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CHAPTER ONE
1.0 INTRODUCTION
Diabetes mellitus is a disease in which blood vessels of glucose (sugar) are high
because the body does not produce or properly use insulin. There are two major forms
of diabetes mellitus. Type 1 diabetes
develops when the pancreas does not produce
insulin. Type 2 diabetes occurs when the body cell resist insulin’s effect (Microsoft
Encarta, 2009). This condition leads to elevated levels of blood glucose. The normal
range of blood glucose level for blood glucose level is between 70-110mg/dl. Insulin is
a hormone that helps to maintain normal blood glucose level by making the body’s cell
absorbs glucose (sugar) so that it can be as a source of energy. In people with diabetes
glucose levels build up in the blood and urine causing excessive urination, thirst,
hunger and problems with fats and protein metabolism because the body cannot
convert glucose into energy, it begins to
break down stored fats for fuel. This
produces increasing amounts of acidic compounds in the blood called ketone bodies
which interfere with cellular respiration energy producing process in cells. Alloxan
induces diabetes mellitus in rats. Alloxan, a beta cytotoxin, induces diabetes in a wide
variety of animal species through damage of insulin secreting cell. In these animals,
with characteristic similar to type 1 diabetes in humans. Hypercholesterolemia and
hypertriglyceridemia are common complications of diabetes mellitus.
(Rerup, C. C.
1999).
Senna tora (originally described by Linne as cassia tora) is a legume in the subfamily
caesalpiniodeae. It grows wild in most of the tropics and is considered a weed in many
1
places. Its native range is not well known but probably South Asia. It is often confused
with Chinese senna or sickle pods obtusifolia. If it is given a distinct common name at
all, it is called sickle wild sensitive plant (nature serve, 200). It has a widely ranging
tropical and the agro climatic conditions, which are conducive for introducing and
domesticating new and exotic plant varieties. The use of the plants, plant extracts and
pure compounds isolated from natural sources provided the foundation to modern
pharmaceutical compounds. An ethno botanical search on fine species senna within
and around Ogbomoso, Oyo state, Nigeria showed their relevance in the local herbal
medicine. In the recent study, screening for hypoglycemic activity of the extract of
senna tora was conducted to provide support for the use of this plant as traditional
medicine. Phytochemical screening provides knowledge of the chemical constituents
of this not only for the discovery of new therapeutic agents, but also for information in
discovering new sources of other materials. The uses of senna tora include the
following, used as liver stimulant, mild laxative, heart tonic, used in treatment of fever,
used to treat eczema and dermatomycosis, etc.
1.1 AIMS AND OBJECTIVES OF THE RESEARCH
Therefore the goal of the study is to:
1.
To determining the blood glucose levels of normal and Alloxan induced
diabetic rats.
2.
To determine the effects of senna tora leaves extract on the blood glucose
levels of the diabetic albino rats.
3.
To compare values before and after induction with Alloxan and senna tora
leaves.
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CHAPTER TWO
2.0 LITERATURE REVIEW
Plant senna tora, formally regarded as cassia tora is God’s gift to man. Senna (from
Arabic Sana), the sennas is a large genus of flowering plants in the family fabaceae,
subfamily caesalpiniodeae. This diverse genus is nature throughout the tropics with
small number of species reaching into temperate regions. The number of species is
usually estimated to be about 360 but believe that there are many as 350.
2.1 SCIENTIFIC CLASSIFICATION OF SENNA TORA
Kingdom: Plantae
Division: Angiospermae
Class: Magnoliopsioda
Sub class: Rosidae
Order: Fabales
Family: Caesalpiniodeae (fabaceae)
Sub family: Caesalpiniodeae
Tribe: Cassieae
Sub tribe: Cassiinae
Genus: Senna
Species: Tora.
Other species in the genus senna include obtusifolia and occidentalis which are similar
both in physical and chemical characteristics and in their applications.
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2.2 DESCRIPTION OF SENNA TORA
The sennas are typically shrubs or sub shrubs, some becoming scandent when
growing into other vegetation. Some are herbs or small trees. Many species have extra
floral nectarines. The leaves are parpinately compound, the leaflets opposite. The
inflorescence is a raceme or some arrangement or racemes. The pedicles lack
bracteoles. The flower lack nectar. They are buzz pollinated and offer pollen as a
reward to pollinators. They stamens may be as few as four, but usually there are ten,
when ten they occur in three sets. The ad axial stamens are staminodial. The four
media stamens are smaller than the three basal stamens. The anthers are basified and
open by two terminal pores or shot slits.
The gynocium is often enantiosylous, which is deflected laterally to the or left. This
makes the flower asymmetrical as well. The fruit is a legume, indehiscent.
2.3 SOME COMMON NAMES OF SENNA TORA
Foetid, cassia, sickle senna, coffee pod, sickle pod, tovaa, and chakvad. Common
Nigeria names include: “ochigichi” (igbos)”ako rere” (Yoruba’s), etc. Due to its
potency in treating diseases and infections (nature serve, 2007).
2.4 GEOGRAPHICAL DISTRIBUTION OF SENNA TORA
Senna
tora
is
worldwide
in
distribution
india,pakiston,bhatan,
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and
very
common
is
Nepal,
Cambodia,Myanmar,Thailand,Vietnam,Indonesia,maylesia,papua,new
guinea,philipines,southwestern
pacific
countries
ranging
from
India
subcontinent(Asian-tropical),Malaysia Solomon islands (the pacific),and Nigeria,
along a stream band in aguodo-okelerin,ogbomoso, where it was found to grow
together with senna occidentalis (ogunkule and ladejobi,2000 and duke,2002).the
countries can be found along the tropical and subtropical regions of the globe at
latitude 10o -30o north and south the equator. The climate consists mostly of the
tropical monsoon climate of 270o (80.60of), 30oc (86of) in summer and 270oc (77.2of)
in winter. The temperature of 24oc (69.8of) in winter, where the rainfall ranging
between 10,795 mm,(425 inches) and 22,907mm (905 inches) as recorded in 1861,
can be expected (aerola,et al.1992). But a temperature of 90c (150f) can be
obtained in Himalayan (nature serve, 2007).
2.5 GROWTH REQUIREMENTS OF SENNA TORA
Senna tora can grow both on sand, loamy, and clay soils and can withstand acidic,
neutral or semi shade or no shade, provided there is moisture. It flowers between the
month of July and September other species flowers within May and June and produces
fruit within august – October. Thus it can grow both in tropic and wastelands in the
cultivated beds and along roadsides (Nature Serve, 2007).
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2.6 PROPAGATION OF SENNA TORA
The seeds of senna tora are scarified and pre-soaked in warm water for 2-3 hours
before sowing usually from early spring to early summer. In a green house (Thompson
and Moran,1989). The seedlings are picked out into individual pots once they are
picked out into individual pots once they are enough to handle. Division is done in the
spring as growth commences (Murray, 1981) and moderate cutting of ripe wood is
done in July, in frames (Huxley, 1992).
2.7 SOME STUDIES ON THE PHYTOCHEMICAL USES OF SENNA TORA
Antioxidant properties (yen and chuang, 2009) and inhibitory effect (Wu et al, 2001)
of the extract of senna tora have already been reported. A recent study was conducted
by nutrition, catholic university of daegu, Korea who conducted that cassia tora
supplements can help improve serum lipid status in type II diabetes without significant
adverse effect (cho et al, 2005). In the recent study, screening for antibacterial as well
as antioxidant activity of the extract of senna tora was conducted to provide support
for the use of this as a traditional medicine.
Phytochemical screening provides knowledge of the chemical constituents of this
plant not only for the discovery of new therapeutic agents but also for information in
discovering new sources of other economic materials. Methanol and aqueous extract
of the dried aerial part of senna tora Roxb. Were subjected to the potential antioxidant
of the extract determined on the basis of their scavenging activity if the stable 1, 1diphenyl-2-prcdrylhdrazyl (DPPH) free radical.ic50 of the methanol extract of senna
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tora possess strong antioxidant activity. However the aqueous both Gram positive and
Gram –negative bacteria in agar diffusion method. The zones of inhibition produced
by the crude methanol and aqueous extract against few sensitive strains were measure
and compared with those of standard antibiotic gentamycin. It is evident that both
extracts are active against both gram positive and gram negative bacteria in agar
diffusion method. The zones of inhibition produced by the crude methanol and
aqueous extract against few sensitive strains were measure and compared with those
of standard antibiotic gentamycin. It is evident that both extracts are active against the
bacteria at low concentrations (uddin,et al 2008).Also the seeds of cassia tora l. have
been conventionally used throughout the American region for several centuries. Its
roasted seeds have a favourable flavour, so it is used popularly as a tea in Korea,
cassia tora l. has been also prescribed in oriental herb medicine to treat night
blindness, hypertension, hypercholesrolemia and constipation. (Ahn.1998., kim et al,
1990), it was reported that cassia tora l.posseses various functional properties
including hypoglycaemic (Lim et al, 1995; Lim and Han, 1997), antimutagenic (Choi
et al, 1997) activities. Cassia tora fiber supplements showed a reduction in the levels
of serum triglycerides and LDL in Korean diabetic patients (Alternative and
Complement Therapies, 2005).
2.8 SOME CHEMICAL PROPERTIES OF SENNA TORA
Cassia tora l. Contains may active substances including nor-rubrofusarin,aurantioobtusin,chryso,obtusin,emoiudin,obrusifolin,chrysophanol,physcion
7
and
chryso-
obtusin-2-o-B-glucoside (Jang et al,2007).the seeds contains anthraquinones and
nanphthopyrones (manila,1998) and also a good source of tannin (gamble,1992).
2.9 USES AND BENEFITS OF CASSIA TORA
-cassia tora is used as coffee substitute.
-It is useful in treating skin diseases like ring worm and itching or body scratch and
psonasis
-the alcoholic or vinegar maceration of pounded fresh leaves is used externally to treat
eczema and dermatomycosis.
-decoction of the fruit is used in treating fever.
It act as a nerve tonic since the herb acts as a kapha and vata dosha suppressant.
-it is consumed in worm infestation and cures the infestation occurring in the body.
-cassia tora acts as a liver stimulant, mild laxative and heart tonic.
-the herb helps in maintaining the normal level of cholesterol.
-it paste is used for treating skin ailments and also for getting rid of chronic diseases.
-its powder proves useful combating indigestion, toning up heart muscles and
purifying blood.
-the juice extracted from its leaves is used in case of skin ailments, rashes and
allergies. It is also used an antidote in case of various poisonings.
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-the leaves and seeds of cassia tora are useful in leprosy, flatulence, cardiac
disorders.(nature serve,2007). They are used in Nepal externally in threading
leucoderma, leprosy and itchy skin (mamandhar, 2002). The seeds are known to
contain anthraguinones and naphthepyrones (mamilla, 1998). They are highly
recommended in agricultural industries in the retrieval of salines, brackish and
alkaline soils. it is used as yellow fertilizer crop in basic soil. Dehydrated seeds are
pre-arranged as protein for livestock feed since it contains about 25% protein (nature
serve, 2007). In the manufacturing industries, the seeds serve as a good source of
tannins (gamble, 1972) and blue, yellow and red dyes. They are also source of cassia
gum and found additives usually used as thickener (nature serve, 2007).
2.10 DEFINITION OF DIABETES
The term diabetes, without qualification, usually refers to diabetes mellitus, which
roughly translates to excessive sweet urine (known as "glycosuria"). Several rare
conditions are also named diabetes. The most common of these is diabetes insipidus
in which large amounts of urine are produced (polyuria), which is not sweet
(insipidus meaning "without taste" in Latin).The two types of diabetes are diabetes
mellitus and diabetes insipidus.
2.11 DIABETES INSIPIDUS
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These are rare diseases caused by deficiency of vasopressin, one of the hormones of
the posterior pituitary gland, which controls the amount of urine secreted by the
kidneys. The symptoms of diabetes insipidus are marked thirst and the excretion of
large quantities of urine, as much as 4 to 10 liters a day. This urine has a low specific
gravity and contains no excess sugar. In many cases, injection or nasal inhalation of
vasopressin controls the symptoms of the disease (Microsoft ® Encarta ® 2009).
2.12 DIABETES MELLITUS
Diabetes mellitus is a disease, in which the pancreas produces insufficient amounts of
insulin, or in which the body’s cells fail to respond appropriately to insulin levels or
poor response to insulin prevents cells from absorbing glucose. Diabetes mellitus is a
common disorder of metabolism in which the body (pancreas) does not produce or
properly use insulin, as result of this, glucose builds up in the blood .when glucoseladen blood passes through the kidneys, the organ that absorb all of the excess
glucose. This excess glucose spills into urine accompanied by water and electrolytesions require by cells to regulate the electric charge and flow of water molecules across
the cell membrane. This causes frequent urination to get rid of the additional water
drawn into the urine; excessive thirst to triggered replacement of lost water and
hunger to replace the glucose lost in urination.
2.13 HISTORY OF DIABETES MELLITUS
10
The term diabetes was coined by Aretaeus of Cappadocia. It was derived from the
Greek verb, diabaínein, itself formed from the prefix dia-, "across, apart," and the verb
bainein, "to walk, stand." The verb diabeinein meant "to stride, walk, or stand with
legs asunder"; hence, its derivative diabetes meant "one that straddles," or specifically
"a compass, siphon." The sense "siphon" gave rise to the use of diabētēs as the name
for a disease involving the discharge of excessive amounts of urine. Diabetes is first
recorded in English, in the form diabetes, in a medical text written around 1425. In
1675, Thomas Willis added the word mellitus, from the Latin meaning "honey", a
reference to the sweet taste of the urine. This sweet taste had been noticed in urine by
the ancient Greeks, Chinese, Egyptians, Indians, and Persians. In 1776, Matthew
Dobson confirmed that the sweet taste was because of an excess of a kind of sugar in
the urine and blood of people with diabetes.
Diabetes mellitus appears to have been a death sentence in the ancient era.
Hippocrates makes no mention of it, which may indicate that he felt the disease was
incurable. Aretaeus did attempt to treat it but could not give a good prognosis; he
commented that "life (with diabetes) is short, disgusting and painful." Sushruta (6th
century BCE) identified diabetes and classified it as Medhumeha. He further
identified it with obesity and sedentary lifestyle, advising exercises to help "cure" it.\
The ancient Indians tested for diabetes by observing whether ants were attracted to
a person's urine, and called the ailment "sweet urine disease" (Madhumeha). The
Chinese, Japanese and Korean words for diabetes are based on the same ideographs
which mean "sugar urine disease".
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In medieval Persia, Avicenna (980–1037) provided a detailed account on diabetes
mellitus in The Canon of Medicine, "describing the abnormal appetite and the
collapse of sexual functions," and he documented the sweet taste of diabetic urine.
Like Aretaeus before him, Avicenna recognized primary and secondary diabetes. He
also described diabetic gangrene, and treated diabetes using a mixture of lupine,
trigonella (fenugreek), and zedoary seed, which produces a considerable reduction in
the excretion of sugar, a treatment which is still prescribed in modern times.
Avicenna also "described diabetes insipidus very precisely for the first time", though
it was later Johann Peter Frank (1745–1821) who first differentiated between
diabetes mellitus and diabetes insipidus.
Although diabetes has been recognized since antiquity, and treatments of various
efficacy have been known in various regions since the Middle Ages, and in legend for
much longer, pathogenesis of diabetes has only been understood experimentally
since about 1900. The discovery of a role for the pancreas in diabetes is generally
ascribed to Joseph von Mering and Oskar Murkowski, who in 1889 found that dogs
whose pancreas was removed developed all the signs and symptoms of diabetes and
died shortly afterwards. In 1910, Sir Edward Albert Sharpey-Schafer suggested that
people with diabetes were deficient in a single chemical that was normally produced
by the pancreas—he proposed calling this substance insulin, from the Latin insula,
meaning island, in reference to the insulin-producing islets of Langerhan s in the
pancreas.
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The endocrine role of the pancreas in metabolism, and indeed the existence of
insulin, was not further clarified until 1921, when Sir Frederick Grant Banting and
Charles Herbert Best repeated the work of Von Mering and Minkowski, and went
further to demonstrate they could reverse induced diabetes in dogs by giving them
an extract from the pancreatic islets of Langerhans of healthy dogs. Banting, Best,
and colleagues (especially the chemist Collip) went on to purify the hormone insulin
from bovine pancreases at the University of Toronto. This led to the availability of an
effective treatment—insulin injections—and the first patient were treated in 1922.
For this, Banting and laboratory director MacLeod received the Nobel Prize in
Physiology or Medicine in 1923; both shared their Prize money with others in the
team who were not recognized, in particular best and Collip. Banting and Best made
the patent available without charge and did not attempt to control commercial
production. Insulin production and therapy rapidly spread around the world, largely
as a result of this decision. Banting is honoured by World Diabetes Day which is held
on his birthday, November 14.The distinction between what is now known as type 1
diabetes and type 2 diabetes was first clearly made by Sir Harold Percival (Harry)
Himsworth. Diabetes is most common in adults over 45 years of age; in people who
are overweight or physically inactive; in individuals who have an immediate family
member with diabetes; and in people of African, Hispanic, and Native American
descent. The highest rate of diabetes in the world occurs in Native Americans. More
women than men have been diagnosed with the disease.
13
In diabetes mellitus low insulin levels or poor response to insulin prevent cells from
absorbing glucose. As a result, glucose builds up in the blood. When glucose-laden
blood passes through the kidneys, the organs that remove blood impurities, the
kidneys cannot absorb all of the excess glucose. This excess glucose spills into the
urine, accompanied by water and electrolytes—ions required by cells to regulate the
electric charge and flow of water molecules across the cell membrane (Microsoft ®
Encarta ® 2009).
2.14 TYPES OF DIABETES MELLITUS
2.15 TYPE1DIABETES MELLITUS: this type of diabetes was formerly called
insulin –dependent diabetes mellitus (IDDM) and juvenile onset diabetes. In this type
of diabetes mellitus, the body (pancreas) fails to produce insulin or produces it only in
very small quantities. Symptoms usually appear suddenly, in individuals under 20
years of age. Most cases occur around puberty around the age of 10-12 years in girls
and 12-14 years in boys. Type 1diabetes is an autoimmune disease, the auto immune
defence system goes awry and attacks healthy tissues that is, the auto immune defence
system against disease is believed to incorrectly identify the islet cells as foreign body
and destroy them. These islet cells are the insulin-producing cells, known as beta cells,
in the pancreas. Insulin dependent diabetes mellitus may be triggered by viruses,
combination of genetic and environmental factors. It can also be caused by surgical
removal if the pancreas. This form of diabetes mellitus requires immediate treatment
by both diet and injections of insulin since it can quickly prove fatal.(Holmsn R.R et
al,1991). This disease condition does not only cause the build up of glucose in the
14
blood but also affects fat metabolism if left untreated. Since there is no conversion of
glucose into energy in the body, it leads to breakdown of body fat as an alternative
source of energy and this leads to production of increasing amount of acidic
compounds in the blood known as ketone which interfere with cellular respiration, the
energy –producing process in cells which can result in coma. Hypercholesterolemia is
a common complication of diabetes is indeed very high, depending on the type of
diabetes and degree of control (Maruf.A, 2007).
TYPE 2 DIABETES MELLITUS: Also known as non0 insulin dependent diabetes
mellitus or adult onset diabetes or adult onset diabetes. It usually affects people of
aged over 40 and progresses gradually. In this type, the pancreas has not ceased to
produce insulin but the hormone (insulin) is not stimulating the glucose uptake in
muscles and tissues required for energy that is the body’s delicate balance between
insulin goes awry. The result is the build up of glucose in blood and urine. Although
the cause of this malfunctioning is unclear, on-insulin dependent diabetes mellitus
tends to run in families. Other risk factors such as increasing age, obesity, and certain
lifestyle probably may contribute to its increased incidence. In developed countries.
(Holman R.R et al, 1991). Symptoms characteristics of type 2 diabetes include
repeated infection or skin sores that heal slowly or not at all, generalised tiredness and
tongling or numbness. In the hands or feet. Non insulin dependent diabetes mellitus
can often be controlled with tablets that reduce the amount of blood glucose.
2.17 GESTATIONAL DIABETES
15
Gestational diabetes mellitus (GDM) resembles type 2 diabetes in several respects,
involving
a
combination
of
relatively
inadequate
insulin
secretion
and
responsiveness. It occurs in about 2%–5% of all pregnancies and may improve or
disappear after delivery. Gestational diabetes is fully treatable but requires careful
medical supervision throughout the pregnancy. About 20%–50% of affected women
develop type 2 diabetes later in life.
Even though it may be transient, untreated gestational diabetes can damage the
health of the foetus or mother. Risks to the baby include macrosomia (high birth
weight), congenital cardiac and central nervous system abnormalities, and skeletal
muscle malformations. Increased foetal insulin may inhibit foetal surficient
production and cause respiratory distress syndrome. Hyperbilirubinemia may result
from red blood cell destruction. In severe cases, prenatal death may occur, most
commonly as a result of poor placental perfusion due to vascular impairment. Labour
induction may be indicated with decreased placental function. A caesarean section
may be performed if there is marked foetal distress or an increased risk of injury
associated with macrosomia, such as shoulder dystocia.
2.18 HYPERGLYCAEMIA AND HYPOGLYCAEMIA
If the body produces too much pituitary hormone or too little insulin, the amount of
sugar in the blood rises abnormally, producing a condition known as hyperglycemia.
In hyperglycemia the blood may contain as much as four times the normal amount of
16
sugar. Hyperglycemia in itself is not lethal, but it is a symptom of a serious disease,
diabetes mellitus. Diabetes is sometimes caused by a tumor or other condition in the
pancreas that prevents the formation of insulin. Diabetic patients do not die of
hyperglycemia, but if they are not given injections of insulin they may die from such
causes as the accumulation of poisons in the body, produced by altered metabolism
of fats; the body of the diabetic consumes fats as a substitute for the sugar that it
cannot use. If an excessive amount of insulin is injected into the body, the amount of
sugar is reduced to a dangerously low level, a condition known as hypoglycemia or
insulin shock. Hypoglycemia, condition characterized by an abnormally low level of
sugar in the blood. Symptoms of hypoglycemia include weakness, shakiness,
nervousness, anxiety, and faintness and actual fainting. Patients also may show
marked personality changes and may seem intoxicated. Hypoglycemia is the result of
hyperinsulinism, or an excess of insulin, due either to an overdose of insulin—in the
case of persons with diabetes mellitus—or to the body's overproduction of insulin.
Insulin is instrumental in regulating carbohydrate metabolism; when hyperinsulinism
occurs, glucose is sharply depleted in the process of conversion to glycogen in the
liver and muscles and to fat in the adipose tissues.
Reactive, or functional, hypoglycemia—the most common type—occurs particularly
among persons under emotional stress. It is also due to overproduction of insulin,
commonly three to five hours after meals. Its symptoms are milder than those
suffered by insulin-dependent diabetics, and it can be controlled by lowering
carbohydrate intake. Because reactive hypoglycemia has many of the classical
17
symptoms of depression or anxiety, it is often wrongly believed to be the cause of
underlying psychological disorders. Even when this physical condition is properly
diagnosed, it is most often found to be incidental to, rather than the direct cause of,
the patient's symptoms (Microsoft ® Encarta ® 2009).
2.19 CAUSES OF DIABETES
The cause of diabetes depends on the type. Type 2 diabetes is due primarily to
lifestyle factors and genetics. Type 1 diabetes is also partly inherited and then
triggered by certain infections, with some evidence pointing at Coxsackie B4 virus.
There is a genetic element in individual susceptibility to some of these triggers which
has been traced to particular HLA genotypes (i.e., the genetic "self" identifiers relied
upon by the immune system). However, even in those who have inherited the
susceptibility, type 1 diabetes mellitus seems to require an environmental trigger.
Some cases of diabetes are caused by the body's tissue receptors not responding to
insulin (even when insulin levels are normal, which is what separates it from type 2
diabetes); this form is very uncommon. Genetic mutations (autosomal or
mitochondrial) can lead to defects in beta cell function. Abnormal insulin action may
also have been genetically determined in some cases. Any disease that causes
extensive damage to the pancreas may lead to diabetes (for example, chronic
pancreatitis and cystic fibrosis). Diseases associated with excessive secretion of
insulin-antagonistic hormones can cause diabetes (which is typically resolved once
18
the hormone excess is removed). Many drugs impair insulin secretion and some
toxins damage pancreatic beta cells. The ICD-10 (1992) diagnostic entity,
malnutrition-related diabetes mellitus (MRDM or MMDM, ICD-10 code E12), was
deprecated by the World Health Organization when the current taxonomy was
introduced in 1999.C:\Users\IZU\Desktop\Downloads\Diabetes_mellitus.htm - cite_noteWHO1999-DefDiagClass-8 list of other causes of diabetes include:




Genetic defects of β-cell Function
o
Maturity onset diabetes of the young (MODY)
o
Mitochondrial DNA mutations
Genetic defects in insulin processing or insulin action
o
Defects in proinsulin conversion
o
Insulin gene mutations
o
Insulin receptor mutations
Exocrine Pancreatic Defects
o
Chronic pancreatitis
o
Pancreatectomy
o
Pancreatic neoplasia
o
Cystic fibrosis
o
Hemochromatosis
o
Fibrocalculous pancreatopathy
Endocrinopathies
o
Growth hormone excess (acromegaly)
19


o
Cushing syndrome
o
Hyperthyroidism
o
Pheochromocytoma
o
Glucagonoma
Infections
o
Cytomegalovirus infection
o
Coxsackievirus B
Drugs
o
Glucocorticoids
o
Thyroid hormone
o
β-adrenergic agonists
2.20 SYMPTOMS OF DIABETES MELLITUS
20
Signs and symptoms
FIG. 1
Overview of the most significant symptoms of diabetes.
The classical symptoms of diabetes are polyuria (frequent urination), polydipsia
(increased thirst) and polyphagia (increased hunger). Symptoms may develop rapidly
(weeks or months) in type 1 diabetes while in type 2 diabetes they usually develop
much more slowly and may be subtle or absent.
Prolonged high blood glucose causes glucose absorption, which leads to changes in
the shape of the lenses of the eyes, resulting in vision changes; sustained sensible
glucose control usually returns the lens to its original shape. Blurred vision is a
21
common complaint leading to a diabetes diagnosis; type 1 should always be
suspected in cases of rapid vision change, whereas with type 2 changes are generally
more gradual, but should still be suspected.
People (usually with type 1 diabetes) may also present with diabetic ketoacidosis, a
state of metabolic dysregulation characterized by the smell of acetone; a rapid, deep
breathing known as Kussmaul breathing; nausea; vomiting and abdominal pain; and
an altered states of consciousness.
A rarer but equally severe possibility is hyperosmolar nonketotic state, which is more
common in type 2 diabetes and is mainly the result of dehydration. Often, the
patient has been drinking extreme amounts of sugar-containing drinks, leading to a
vicious circle in regard to the water loss. A number of skin rashes can occur in
diabetes that are collectively known as diabetic dermadromes.
2.21 PATHOPHYSIOLOGY OF DIABETES MELLITUS
Insulin is the principal hormone that regulates uptake of glucose from the blood into
most cells (primarily muscle and fat cells, but not central nervous system cells).
Therefore deficiency of insulin or the insensitivity of its receptors plays a central role
in all forms of diabetes mellitus.
Humans are capable of digesting some carbohydrates, in particular those most
common in food; starch, and some disaccharides such as sucrose, are converted
22
within a few hours to simpler forms most notably the monosaccharide glucose, the
principal carbohydrate energy source used by the body. The most significant
exceptions are fructose, most disaccharides (except sucrose and in some people
lactose), and all more complex polysaccharides, with the outstanding exception of
starch. The rest are passed on for processing by gut flora largely in the colon. Insulin
is released into the blood by beta cells (β-cells), found in the Islets of Langerhans in
the pancreas, in response to rising levels of blood glucose, typically after eating.
Insulin is used by about two-thirds of the body's cells to absorb glucose from the
blood for use as fuel, for conversion to other needed molecules, or for storage.
Insulin is also the principal control signal for conversion of glucose to glycogen for
internal storage in liver and muscle cells. Lowered glucose levels result both in the
reduced release of insulin from the beta cells and in the reverse conversion of
glycogen to glucose when glucose levels fall. This is mainly controlled by the
hormone glucagon which acts in the opposite manner to insulin. Glucose thus
forcibly produced from internal liver cell stores (as glycogen) re-enters the
bloodstream; muscle cells lack the necessary export mechanism. Normally liver cells
do this when the level of insulin is low (which normally correlates with low levels of
blood glucose).
Higher insulin levels increase some anabolic ("building up") processes such as cell
growth and duplication, protein synthesis, and fat storage. Insulin (or its lack) is the
principal signal in converting many of the bidirectional processes of metabolism from
23
a catabolic to an anabolic direction, and vice versa. In particular, a low insulin level is
the trigger for entering or leaving ketosis (the fat burning metabolic phase).
If the amount of insulin available is insufficient, if cells respond poorly to the effects
of insulin (insulin insensitivity or resistance), or if the insulin itself is defective, then
glucose will not have its usual effect so that glucose will not be absorbed properly by
those body cells that require it nor will it be stored appropriately in the liver and
muscles. The net effect is persistent high levels of blood glucose, poor protein
synthesis, and other metabolic derangements, such as acidosis.
When the glucose concentration in the blood is raised beyond its renal threshold
(about 10 mmol/L, although this may be altered in certain conditions, such as
pregnancy), reabsorption of glucose in the proximal renal tubuli is incomplete, and
part of the glucose remains in the urine (glycosuria). This increases the osmotic
pressure of the urine and inhibits reabsorption of water by the kidney, resulting in
increased urine production (polyuria) and increased fluid loss. Lost blood volume will
be replaced osmotically from water held in body cells and other body compartments,
causing dehydration and increased thirst.
2.22 DIAGNOSIS, TREATMENT AND MANAGEMENT OF DIABETES
MELLITUS
Diabetes mellitus is a chronic disease which is difficult to cure. Management
concentrates on keeping blood sugar levels as close to normal ("euglycemia") as
24
possible without presenting undue patient danger. Diabetes is detected by measuring
the amount of glucose in the blood after an individual has fasted (abstained from food)
for about eight hours. In some cases, physicians diagnose diabetes by administering an
oral glucose tolerance test, which measures glucose levels before and after a specific
amount of sugar has been ingested.
Once diabetes is diagnosed, treatment consists of controlling the amount of glucose
in the blood and preventing complications. Depending on the type of diabetes, this
can be accomplished through regular physical exercise, a carefully controlled diet,
and medication. Individuals with Type 1 diabetes must receive insulin, often two to
four times a day, to provide the body with the hormone it does not produce. Insulin
cannot be taken orally, because it is destroyed in the digestive system. Consequently,
insulin-dependent diabetics have historically injected the drug using a hypodermic
needle or a beeper-sized pump connected to a needle inserted under the skin. In
2006 the United States Food and Drug Administration approved a form of insulin that
can be inhaled and then is absorbed by blood in the lungs.
The amount of insulin needed varies from person to person and may be influenced by
factors such as a person’s level of physical activity, diet, and the presence of other
health disorders. Typically, individuals with Type 1 diabetes use a meter several times
a day to measure the level of glucose in a drop of their blood obtained by pricking a
fingertip. They can then adjust the dosage of insulin, physical exercise, or food intake
to maintain the blood sugar at a normal level. People with Type 1 diabetes must
carefully control their diets by distributing meals and snacks throughout the day so as
25
not to overwhelm the ability of the insulin supply to help cells absorb glucose. They
also need to eat foods that contain complex sugars, which break down slowly and
cause a slower rise in blood sugar levels. Generally, insulin is self-administered by
patients by injection or with automatic drug injectors containing a cartridge of insulin
can be carried in the pocket for erase and speed of treatment. (muherjee S.K,1990)
herbal
medicine
can
be
used
to
treat
diabetes
mellitus
(kamseswara
R,1997).Although most persons with Type 1 diabetes strive to lower the amount of
glucose in their blood, levels that are too low can also cause health problems. For
example, if a person with Type 1 diabetes takes too much insulin, it can produce low
blood sugar levels. This may result in hypoglycemia, a condition characterized by
shakiness, confusion, and anxiety. A person who develops hypoglycemia can combat
symptoms by ingesting glucose tablets or by consuming foods with high sugar
content, such as fruit juices or hard candy. (Microsoft ® Encarta ® 2009). Oral
medications may be used in the case of type 2 diabetes, as well as insulin).
For persons with Type 2 diabetes, treatment begins with diet control, exercise, and
weight reduction, although over time this treatment may not be adequate. People with
Type 2 diabetes typically work with nutritionists to formulate a diet plan that regulates
blood sugar levels so that they do not rise too swiftly after a meal. A recommended
meal is usually low in fat (30 percent or less of total calories), provides moderate
protein (10 to 20 percent of total calories), and contains a variety of carbohydrates,
such as beans, vegetables, and grains. Regular exercise helps body cells absorb
glucose—even ten minutes of exercise a day can be effective. Diet control and
26
exercise may also play a role in weight reduction, which appears to partially reverse
the body’s inability to use insulin.
For some people with Type 2 diabetes, diet, exercise, and weight reduction alone
may work initially, but eventually this regimen does not help control high blood sugar
levels. In these cases, oral medication may be prescribed. If oral medications are
ineffective, a person with Type 2 diabetes may need insulin doses or a combination
of oral medication and insulin. About 50 percent of individuals with Type 2 diabetes
require oral medications, 40 percent require insulin or a combination of insulin and
oral medications, and 10 percent use diet and exercise alone.
2.23 LIFESTYLE MODIFICATIONS
There are roles for patient education, dietetic support, sensible exercise, with the goal
of keeping both short-term and long-term blood glucose levels within acceptable
bounds. In addition, given the associated higher risks of cardiovascular disease,
lifestyle modifications are recommended to control blood pressure in patients with
hypertension, cholesterol in those with dyslipidemia, as well as exercising more,
smoking less or ideally not at all, consuming a recommended diet. Patients with foot
problems are also recommended to wear diabetic socks, and possibly diabetic shoes
2.24 EFFECT AND COMPLICATIONS OF DIABETES MELLITUS
If left untreated, diabetes mellitus may cause life-threatening complications. Type 1
diabetes can result in diabetic coma (a state of unconsciousness caused by extremely
27
high levels of glucose in the blood) or death. In both Type 1 and Type 2 diabetes,
complications may include blindness, kidney failure, and heart disease. Diabetes can
cause tiny blood vessels to become blocked; when this occurs in blood vessels of the
eye, it can result in retinopathy (the breakdown of the lining at the back of the eye),
causing blindness. Diabetes mellitus is the leading cause of new cases of blindness in
people aged 20 to 74. In the kidneys, diabetes can lead to nephropathy (the inability
of the kidney to properly filter toxins from the blood). About 40 percent of new cases
of end-stage renal disease (kidney failure) are caused by diabetes mellitus. Blockages
of large blood vessels in diabetics can lead to many cardiovascular problems,
including high blood pressure, heart attack, and stroke. Although these conditions
also occur in non-diabetic individuals, people with diabetes are two to four times
more likely to develop cardiovascular disorders.
Diabetes mellitus may also cause loss of feeling, particularly in the lower legs. This
numbness may prevent a person from feeling the pain or irritation of a break in the
skin or of foot infection until after complications have developed, possibly
necessitating amputation of the foot or leg. Burning pain, sensitivity to touch, and
coldness of the foot, conditions collectively known as neuropathy, can also occur.
Other complications include higher-risk pregnancies in diabetic women and a greater
occurrence of dental disease.
2.25 ALLOXAN
28
Alloxan is an oxidation product of uric acid that is found in the human intestine in
diarrhoea. Alloxan has been used to produce diabetes in experimental animals by
destroying the insulin-secreting islets cells of the pancreas. Alloxan (2, 4, 5, 6tetraoxypyridine; 2, 4, 5, 6-pyrimidineterone) is an oxygenated pyrimidine derivative.
It is present as Alloxan hydrate in an aqueous solution.
2.26 HISTORY OF ALLOXAN
Alloxan is a toxic glucose analogue, which selectively destroys insulin producing cell
in the pancreas when administered to rodents and diabetes mellitus (called “Alloxan
diabetes”). In those animals, with characteristics similar to type 1 diabetes in humans,
Alloxan is selectively toxic to insulin-producing pancreatic beta cells because it
preferentially accumulates in beta cells through uptake via the GLUT2 glucose
transporter, Alloxan in the presence of intracellular thiols, generates reactive oxygen
species (ROS) in a cyclic reaction with its reduction product, dialuric acid. The beta
cells toxic action of Alloxan is initiated by free radicals formed in this redox reaction.
2.27 DISCOVERY OF ALLOXAN
The compound was discovered by Justus van Liebig and Friedrich Wohler following
the discovery of urea in 1928 and is one of the oldest name organic compounds that
exist.
2.28 SYNTHESIS OF ALLOXAN
29
The original properties for Alloxan were by oxidation of uric by nitric acid. In other
method the monodyrate is prepared by oxidation of baribituric acid by chromium
trioxide. Alloxan is a strong oxidizing agent and it forms a hemiacetal with its
reduced reaction product dialuric acid (in which a carbonyl group is group is reducted
to a hydroxyl group) which is called alloxantin.
2.29 STRUCTURE OF ALLOXAN
Alloxan or mesoxalylurea is an organic compound based on a pyrimidine heterocyclic
skeleton. This compound has a high affinity for water and therefore exists as the
monohydrate.
IUPAC name: 1, 3-diaziane-2-4-5-6-tetrone.
FIG. 2
Alloxan has a molecular formular:4H4N2O5. Molar mass 160.09.melting
point:2530c.
2.30 IMPACT UPON BETA CELLS
Because it selectively kills the insulin-producing beta cells found in the pancreas.
Alloxan is used to induce diabetes in laboratory animals. This occurs most likely
30
because of selective uptake of the compound due to its structural similarity to glucose
as well as the beta cell’s highly efficient uptake mechanism.
2.31 MECHANISM OF ALLOXAN ACTION
The precise mechanism of the selective beta- cytotoxicity of Alloxan is not properly
understood. Coopertein Watkins and lazarow (1964) have known that Alloxan
probably exerts toxic effect on the beta-cells by selectively interacting with certain
components of the plasma membrane. This results in an altered permeability which
permits the diffusion of extra cellular fluid markers such as D-mannitol and insulin
into the surrounding incubation medium. However, the fact that only beta-cell
components of intermediate cells are affected by Alloxan suggests that there is no
comparable damage to their plasma membrane as occurs in the beta-cell (cooperstein
S, et al, 1964),but is consistent with the possibility that Alloxan interacts with the
membranes of the beta-cell cytoplasmic organelle. In intermediate cells this imitates
the remarkably selective recognition and an autophary of these organelles. The same
considerations probably apply to the effects of streptozotoxia on the intermediate
cells. Alloxan is known to induce diabetic renal changes as well as causing
nephrotoxic alterations however. No ultra structural study has been performed to
differenciate diabetic verses toxic effect of tubules and glomerulus (Andrew p, 1992)
31
2.30 PANCREAS
Pancreas, conglomerate gland lying transversely across the posterior wall of the
abdomen. It varies in length from 15 to 20 cm (6 to 8 in) and has a breadth of about
3.8 cm (about 1.5 in) and a thickness of from 1.3 to 2.5 cm (0.5 to 1 in). Its usual
weight is about 85 gm (about 3in), and its head lies in the concavity of the
duodenum.
The pancreas has both an exocrine and an endocrine secretion. The exocrine
secretion is made up of a number of enzymes that are discharged into the intestine
to aid in digestion. The endocrine secretion, insulin, is important in the metabolism
of sugar in the body Insulin is produced in small groups of especially modified
glandular cells in the pancreas; these cell groups are known as the islets of
Langerhans. The failure of these cells to secrete sufficient amounts of insulin causes
diabetes
In 1968 a team of surgeons at the medical school of the University of Minnesota
performed the first pancreas transplants on four diabetics, using the pancreases of
cadavers. Since this first transplant, thousands of pancreas transplants have been
performed in the United States. According to the International Pancreas Transplant
Registry (IPTR), based at the University of Minnesota, about 14,705 pancreas
transplant operations occurred from 1987 to 2003. The majority of these, about 78
percent, were simultaneous pancreas-kidney transplants, known as SPK transplants.
These transplants are performed because diabetics also suffer kidney failure. About
32
14 percent of pancreas transplants are known as pancreas after kidney (PAK)
transplants, indicating that a kidney transplant was performed prior to the pancreas
transplant. A smaller percentage of transplants, about 6 percent, are pancreas
transplants alone (PTA).
Patient survival after one year for transplants performed from 1999 to 2003 was at
least 94 percent in all categories and was highest, at 98.4 percent, in the PTA
category, according to the IPTR. The ten-year survival rate, based on data from 1987
to 1992, was 68 percent for PTA transplants, 63 percent for SPK transplants, and 54
percent for PAK transplants. Patients who have pancreas transplants must take
immunosuppressant drugs, such as cyclosporine, for the remainder of their lives. The
pancreas has both a digestive and a hormonal function. Composed mainly of
exocrine tissue, it secretes enzymes into the small intestine, where they help break
down fats, carbohydrates, and proteins. Pockets of endocrine cells called the islets of
Langerhans produce glucagon and insulin, hormones that regulate blood-sugar level.
(Encarta Encyclopaedia)
33
FIG. 4
FIG.5
2.33THE STRUCTURES OF PANCREAS
34
Diseases of the pancreas are relatively rare. Cancer of the pancreas is rare but
deadly. It is the fourth leading cause of cancer death in the United States and the
fifth leading cause worldwide. The mortality rate is high because pancreatic cancer
produces few if any symptoms and so is often not detected until it has spread to
other organs. Smoking is thought to cause about 30 percent of pancreatic cancers.
Haemorrhage in the pancreas and acute pancreatitis are also serious conditions. If
not treated quickly, they may cause death. The symptoms are not definite,
resembling those of peritonitis or intestinal obstruction.
2.34 INSULIN
Insulin is a polypeptide hormone formed after eliminate after elimination of (peptide
by hydro sis of chains 21 and 30 amino acids) connected by two disulfide bridges. it is
secreted by the b cells of the islets of langerhams of the pancreas and exerts an
hypoglycaemic action. It belongs to the group of peptide called IGF (insulin like
growth factors) or somatomedins.
2.35 HISTORY OF INSULIN
Insulin was first extracted from the pancreatic tissue of dogs in 1921 by the Canadian
physiologists Sir Frederick Grant Banting and Charles Herbert Best and the British
physiologist John James Rickard Macleod. The Canadian biochemist James Bertram
Collip then produced it in sufficiently pure form to be injected into humans. The
molecular structure of insulin was determined in 1955 by the British biochemist
Frederick Sanger; it was the first protein to be deciphered. Human insulin, the first
human protein to be synthesized, was made in 1965. In 1981 insulin made in bacteria
35
by genetic engineering became the first human hormone obtained in this way to be
used to treat human disease. (Microsoft ® Encarta ® 2009).
2.36 THE STRUCTURE OF INSULIN
FIG. 6
2.37 INSULIN BIOSYNTHESIS
Insulin is produced by the B cells of the islets of Langerhans in the pancreas. As is
usual with secretory proteins, the hormone’s precursor (pre proinsulin) carries a signal
peptide that directs the peptide chain to the interior of the endoplasmic reticulum is
produced in the ER by cleavage of the signal peptide and formation of disulfide
bonds. Proinsulin passes to the Golgi apparatus, where it is packed into vesicles
granules. After cleavage of the C peptide, mature insulin is formed in the granules and
is stored in the form of zinc-containing hexamers until secretion.
2.38 EFFECTS OF INSULIN DEFICIENCY
36
The effects of insulin deficiency are mainly found on carbohydrate and fats
metabolism. The effects of insulin on carbohydrate metabolism can be described as
stimulation of glucose utilization and inhibition of gluconeogenesis. In addition, the
transport of glucose from the blood into most tissues is also insulin-dependent
(exceptions to this include the liver, CNS, and erythrocytes).The lipid metabolism of
adipose tissue is also influenced by the hormone. In these cells, insulin stimulates the
reorganization of glucose into fatty acids. This is mainly based on activation of acetyl
CoA carboxylase and increased availability of NADPH+H+ due to increased PPP
activity On the other hand, insulin also inhibits the degradation of fat by hormone
sensitive. Hormone sensitive lipases and prevents the breakdown of muscle protein.
In muscle and adipose tissue – the two most important glucose consumers—glucose
uptake and glucose utilization are impaired by insulin deficiency. Glucose utilization
in the liver is also reduced. At the same time, gluconeogenesis is stimulated, partly
due to increased proteolysis in the muscles. This increases the blood sugar level still
further. When the capacity of the kidneys to resorb glucose is exceeded (at plasma
concentrations of 9 mM or more), glucose is excreted in the urine (glucosuria). The
increased degradation of fat that occurs in insulin deficiency also has serious effects.
Some of the fatty acids that accumulate in large quantities are taken up by the liver
and used for lipoprotein synthesis (hyperlipidemia), and the rest are broken down into
acetyl CoA. As the tricarboxylic acid cycle is not capable of taking up such large
quantities of acetyl CoA, the excess is used to form ketone bodies (acetoacetate and
hydroxybutyrate As H+ ions are released in this process, diabetics not receiving
adequate treatment can suffer severe metabolic acidosis (diabetic coma). The acetone
37
that is also formed gives these patients’ breath a characteristic odour. In addition,
large amounts of ketone body anions appear in the urine (ketonuria).
2.39 FAT METABOLISM
Fat metabolism in adipose tissue (top). Fats (triacylglycerols) are the most important
energy reserve in the animal organism. They are mostly stored in insoluble form in the
cells of adipose tissue—the adipocytes—where they are constantly being synthesized
and broken down again. As precursors for the biosynthesis of fats.
(Lipogenesis), the adipocytes use triacylglycerols from lipoproteins (VLDLs and
chylomicrons; which are formed in the liver and intestines and delivered by the blood.
Lipoprotein lipase, which is located on the inner surface of the blood capillaries,
cleaves these triacylglcerols into glycerol and fatty acids, which are taken up by the
adipocytes and converted back into fats.
The degradation of fats (lipolysis) is catalyzed in adipocytes by hormone-sensitive
lipase an enzyme that is regulated by various hormones by cAMP-dependent
interconversion. The amount of fatty acids released depends on the activity of this
lipase; in this way, the enzyme regulates the plasma levels of fatty acids. In the blood
plasma, fatty acids are transported in free form-i. e., non-esterified. Only short-chain
fatty acids are soluble in the blood; longer, less water-soluble fatty acids are
transported bound to albumin.
2.40 DEGRADATION OF FATTY ACIDS IN THE LIVER.
38
Many tissues take up fatty acids from the blood plasma in order to synthesize fats or to
obtain energy by oxidizing them. The metabolism of fatty acids is particularly
intensive in the hepatocytes in the liver. The most important process in the
degradation of fatty acids is -oxidation—a metabolic pathway in the mitochondrial
matrix Initially, the fatty acids in the cytoplasm are activated by binding to coenzyme
A into acyl CoA .Then, with the help of a transport system (the carnitine shuttle the
activated fatty acids enter the mitochondrial matrix, where they are broken down into
acetyl CoA. The resulting acetyl residues can be oxidized to CO2 in the tricarboxylic
acid cycle, producing reduced coenzyme and ATP derived from it by oxidative
phosphorylation. If acetyl CoA production exceeds the energy requirements of the
hepatocytes as is the case when there is a high level of fatty acids in the blood plasma
(typically in hunger and diabetes mellitus) then the excess is converted into ketone
bodies .These serve exclusively to supply other tissues with energy.
2.41 FAT SYNTHESIS IN THE LIVER.
Fatty acids and fats are mainly synthesized in the liver and in adipose tissue, as well
as in the kidneys, lungs, and mammary glands. Fatty acid biosynthesis occurs in the
cytoplasm in contrast to fatty acid degradation. The most important precursor is
glucose, but certain amino acids can also be used. The first step is carboxylation of
acetyl CoA to malonyl CoA. This reaction is catalyzed by acetyl-coA carboxylase,
which is the key enzyme in fatty acid biosynthesis. Synthesis into fatty acids is carried
out by fatty acid synthase. This multifunctional enzyme starts with one molecule of
39
acetyl- CoA and elongates it by adding malonyl groups in seven reaction cycles until
palmitate is reached. One CO2 molecule is released in each reaction cycle. The fatty
acid therefore grows by two carbon units each time. NADPH+H are used as the
reducing agent and are derived either from the pentose phosphate pathway or from
isocitrate dehydrogenase and malic enzyme reactions. The elongation of the fatty acid
by fatty acid synthase concludes at C16, and the product, palmitate, is released.
Unsaturated fatty acids and long-chain fatty acids can arise from palmitate in
subsequent reactions. Fats are finally synthesized from activated fatty acids (acyl
CoA) and glycerol 3-phosphate. To supply peripheral tissues, fats are packed by the
hepatocytes into lipoprotein complexes of the VLDL type and released into the blood
in this form.
Glucose is the universal fuel for human cells. Every cell type in the human is able to
generate adenosine triphosphate (ATP) from glycolysis, the pathway in which glucose
is oxidized and cleaved to form pyruvate. The importance of glycolysis in our fuel
economy is related to the availability of glucose in the blood, as well as the ability of
glycolysis to generate ATP in both the presence and absence of O2. Glucose is the
major sugar in our diet and the sugar that circulates in the blood to ensure that all cells
have a continuous fuel supply. The brain uses glucose almost exclusively as a fuel.
2.42 GLYCOLYSIS
Glycolysis begins with the phosphorylation of glucose to glucose 6-phosphate
(glucose-6-P) by hexokinase (HK). In subsequent steps of the pathway, one glucose6-P molecule is oxidized to two pyruvate molecules with generation of two molecules
40
of NADH.A new generation of two molecules of ATP occurs through direct transfer
of high-energy phosphate from intermediates of the pathway to ADP (substrate level
phosphorylation). Glycolysis occurs in the cytosol and generates cytosolic NADH.
Because NADH cannot cross the inner mitochondrial membrane, its reducing
equivalents are transferred to the electron transport chain by either the malateaspartate shuttle or the glycerol 3 phosphate shuttle Pyruvate is then oxidized
completely to CO2 by pyruvate dehydrogenase and the TCA cycle. Complete aerobic
oxidation of glucose to CO2 can generate approximately 30 to 32 moles of ATP per
mole of glucose. When cells have a limited supply of oxygen (e.g., kidney medulla),
or few or no mitochondria (e.g., the red cell), or greatly increased demands for ATP
(e.g., skeletal muscle during high intensity exercise), they rely on anaerobic glycolysis
for generation of ATP. In anaerobic glycolysis, lactate dehydrogenase oxidizes the
NADH generated from glycolysis by reducing pyruvate to lactate because O2 is not
required to reoxidize the NADH; the pathway is referred to as anaerobic. The energy
yield from anaerobic glycolysis (2 moles of ATP per mole of glucose) is much lower
than the yield from aerobic oxidation. The lactate (lactic acid) is released into the
blood. Under pathologic conditions that cause hypoxia, tissues may generate enough
lactic acid to cause lactic acidemia. In each cell, glycolysis is regulated to ensure that
ATP homeostasis is maintained, without using more glucose than necessary. In most
cell types, hexokinase (HK), the first enzyme of glycolysis, is inhibited by glucose 6phosphate. Thus, glucose is not taken up and phosphorylated by a cell unless glucose6-P enters a metabolic pathway, such as glycolysis or glycogen synthesis. The control
of glucose-6-P entry into glycolysis occurs at phosphofructokinase-1(PFK-1), the rate-
41
limiting enzyme of the pathway. PFK-1 is allosterically inhibited by ATP and
allosterically activated by AMP. AMP increases in the cytosol as ATP is hydrolyzed
by energy-requiring reactions. Living cells require a constant source of fuels from
which to derive ATP for the maintenance of normal cell function and growth.
Therefore, a balance must be achieved between carbohydrate, fat, and protein intake,
their storage when present in excess of immediate need, and their mobilization and
synthesis when in demand. The balance between need and availability is referred to as
metabolic homeostasis. The inter tissue integration required for metabolic homeostasis
is achieved in three principal ways: The concentration of nutrients or metabolites in
the blood affects the rate at which they are used and stored in different tissues.
Glucose
Glucose-6 -phosphate
Fructose-6-phosphate
Fructose-1, 6-diphosphate
Dihydroxy
Glyceraldehyde-3-phosphate
Acetone
42
Phosphate
1,3-diphosphoglycerate (2)
3-phosphoglycerate (2)
2-phosphoglycerate (2)
Phosphoenolpyruvate (2)
Pyruvate
2.43 CARBOHYDRATE METABOLISM
Glucose is central to all of metabolism. It is the universal fuel for human cells and the
source of carbon for the synthesis of most other compounds. Every human cell type
uses glucose to obtain energy. The release of insulin and glucagon by the pancreas
aids in the body’s use and storage of glucose. Other dietary sugars (mainly fructose
and galactose) are converted to glucose or to intermediates of glucose metabolism.
Glucose is the precursor for the synthesis of an array of other sugars required for the
production of specialized compounds, such as lactose, cell surface antigens,
nucleotides, or glycosaminoglycans. Glucose is also the fundamental precursor of
noncarbohydrate compounds; it can be converted to lipids (including fatty acids,
cholesterol, and steroid hormones), amino acids, and nucleic acids. Only those
compounds that are synthesized from vitamins, essential amino acids, and essential
fatty acids cannot be synthesized from glucose in humans. More than 40% of the
calories in the typical diet in the United States are obtained from starch, sucrose, and
lactose. These dietary carbohydrates are converted to glucose, galactose, and fructose
43
in the digestive tract. Monosaccharides are absorbed from the intestine, enter the
blood, and travel to the tissues where they are metabolized. After glucose is
transported into cells, it is phosphorylated by a hexokinase to form glucose 6phosphate. Glucose 6-phosphate can then enter a number of metabolic pathways. The
three that are common to all cell types are glycolysis, the pentose phosphate pathway,
and glycogen synthesis. In tissues, fructose and galactose are converted to
intermediates of glucose metabolism. Thus, the fate of these sugars parallels that of
glucose.The major fate of glucose 6-phosphate is oxidation via the pathway of
glycolysis, which provides a source of ATP for all cell types. Cells that lack
mitochondria cannot oxidize other fuels. They produce ATP from anaerobic
glycolysis (the conversion of glucose to lactic acid). Cells that contain mitochondria
oxidize glucose to CO2 and H2O via glycolysis and the TCA cycle. Some tissues,
such as the brain, depend on the oxidation of glucose to CO2 and H2O for energy
because they have a limited capacity to use other fuels. Glucose produces the
intermediates of glycolysis and the TCA cycle that are used for the synthesis of amino
acids and both the glycerol and fatty acid moieties of triacylglycerols. Another
important fate of glucose 6-phosphate is oxidation via the pentose phosphate pathway,
which generates NADPH. The reducing equivalents of NADPH are used for
biosynthetic reactions and for the prevention of oxidative damage to cells in this
pathway; glucose is oxidatively decarboxylated to 5-carbon sugars (pentoses), which
may re-enter the glycolytic pathway. They also may be used for nucleotide synthesis.
There are also non-oxidative reactions, which can convert six- and five-carbon sugars.
44
2.44 METABOLIC HOMEOSTASIS
Living cells require a constant source of fuels from which to derive ATP for the
maintenance of normal cell function and growth. Therefore, a balance must be
achieved between carbohydrate, fat, and protein intake, their storage when present in
excess of immediate need, and their mobilization and synthesis when in demand. The
balance between need and availability is referred to as metabolic homeostasis. The
intertissue integration required for metabolic homeostasis is achieved in three
principal ways:
• The concentration of nutrients or metabolites in the blood affects the rate at which
they are used and stored in different tissues.
• Hormones carry messages to individual tissues about the physiologic state of the
body and nutrient supply or demand.
• The central nervous system uses neural signals to control tissue metabolism, directly
or through the release of hormones. Insulin and glucagon are the two major hormones
that regulate fuel storage and mobilization. Insulin is the major anabolic hormone of
the body. It promotes the storage of fuels and the utilization of fuels for growth.
Glucagon is the major hormone of fuel mobilization. Other hormones, such as
epinephrine, are released as a response of the central nervous system to hypoglycemia,
exercise, or other types of physiologic stress. Epinephrine and other stress hormones
also increase the availability of fuels. The special role of glucose in metabolic
homeostasis is dictated by the fact that many tissues (e.g., the brain, red blood cells,
the lens of the eye, the kidney medulla, exercising skeletal muscle) are dependent on
glycolysis for all or a portion of their energy needs and require uninterrupted access to
45
glucose on a second-to-second basis to meet their rapid rate of ATP utilization. In the
adult, a minimum of 190 g glucose is required per day; approximately 150 g for the
brain and 40 g for other tissues. Significant decreases of blood glucose below 60
mg/dL limit glucose metabolism in the brain and elicit hypoglycemic symptoms (as
experienced by Bea Selmass), presumably because the overall process of glucose flux
through the blood-brain barrier, into the interstitial fluid, and subsequently into the
neuronal cells, is slow at low blood glucose levels because of the Km values of the
glucose transporters required for this to occur. The continuous movement of fuels into
and out of storage depots is necessitated by the high amounts of fuel required each day
to meet the need for ATP. Disastrous results would occur if even a day’s supply of
glucose, amino acids, and fatty acids were left circulating in the blood. Glucose and
amino acids would be at such high concentrations that the hyperosmolar effect would
cause progressively severe neurologic deficits and even coma. The concentration of
glucose and amino acids would be above the renal tubular threshold for these
substances (the maximal concentration in the blood at which the kidney can
completely resorb metabolites), and some of these compounds would be wasted as
they spilled over into the urine. Nonenzymatic glycosylation of proteins would
increase at higher blood glucose levels. Triacylglycerols circulate in cholesterolcontaining lipoproteins, and the levels of these lipoproteins would be chronically
elevated, increasing the likelihood of atherosclerotic vascular disease. Consequently,
glucose and other fuels are continuously moved in and out of storage depots as
needed.
46
2.45 MAJOR HORMONES OF METABOLIC HOMEOSTASIS
The hormones that contribute to metabolic homeostasis respond to changes in the
circulating levels of fuels that, in part, are determined by the timing and composition
of our diet. Insulin and glucagon are considered the major hormones of metabolic
homeostasis because they continuously fluctuate in response to our daily eating
pattern. They provide good examples of the basic concepts of hormonal regulation.
Certain features of the release and action of other insulin counter regulatory
hormones, such as epinephrine, nor epinephrine, and cortisol, will be described and
compared with insulin and glucagon. Insulin is the major anabolic hormone that
promotes the storage of nutrients: glucose storage as glycogen in liver and muscle,
conversion of glucose to triacylglycerols in liver and their storage in adipose tissue
and amino acid uptake and protein synthesis in skeletal muscle. It also increases the
synthesis of albumin and other blood proteins by the liver. Insulin promotes the
utilization of glucose as a fuel by stimulating its transport into muscle and adipose
tissue. At the same time, insulin acts to inhibit fuel mobilization. Glucagon acts to
maintain fuel availability in the absence of dietary glucose by stimulating the release
of glucose from liver glycogen, by stimulating gluconeogenesis from lactate, glycerol,
and amino acids, and, in conjunction with decreased insulin, by mobilizing fatty acids
from adipose triacylglycerols to provide an alternate source of fuel. Its sites of action
are principally the liver and adipose tissue; it has no influence on skeletal muscle
metabolism because muscle cells lack glucagon receptors. The release of insulin from
the beta cells of the pancreas is dictated primarily by the blood glucose level. The
highest levels of insulin occur approximately 30 to 45 minutes after a high-
47
carbohydrate meal. They return to basal levels as the blood glucose concentration
falls, approximately 120 minutes after the meal. The release of glucagon from the
alpha cells of the pancreas, conversely, is controlled principally through suppression
by glucose and insulin. Therefore, the lowest levels of glucagon occur after a highcarbohydrate meal. Because all of the effects of glucagon are opposed by insulin, the
simultaneous stimulation of insulin release and suppression of glucagon secretion by a
high carbohydrate meal provides an integrated control of carbohydrate, fat, and
protein metabolism.
2.46 CHANGES IN HORMONE LEVELS AFTER A MEAL
After a typical high carbohydrate meal, the pancreas is stimulated to release the
hormone insulin, and release of the hormone glucagon is inhibited circle. Endocrine
hormones are released from endocrine glands, such as the pancreas, in response to a
specific stimulus. They travel in the blood, carrying messages between tissues
concerning the overall physiologic state of the body. At their target tissues, they adjust
the rate of various metabolic pathways to meet the changing conditions. The
endocrine hormone insulin, which is secreted from the pancreas in response to a highcarbohydrate meal, carries the message that dietary glucose is available and can be
used and stored. The release of another hormone, glucagon, is suppressed by glucose
and insulin. Glucagon carries the message that glucose must be generated from
endogenous fuel stores. The subsequent changes in circulating hormone levels cause
Changes in the body’s metabolic patterns, involving a number of different tissues and
metabolic pathways.
48
2.47 CHANGES IN HORMONES LEVEL DURING DIABETES MELLITUS
Diabetes mellitus is associated little or low levels of insulin with high levels of
glucagon. Since the pancreas cannot be able to release insulin or inability of the body
cells to respond to the insulin produced, the level of glucagon increases to maintain
the availability of energy (fuel) in the body system.
CHAPTER THREE
3.0 MAREIALS AND METHODS
3.1 PLANT MATERIALS
The fresh leaves of senna tora were dried and used for this study. The fresh leaves
were harvested from “Ugba” village in Amorji-Nike Emene very close to caritas
university Amorji -Nike Enugu and were identified by the villagers and also by Mr.
Moses Ezenwali of department of biochemistry, Caritas University Amorji- nike.
3.2 ANIMALS
A total number of twenty (20) albino rats of either sex weighing135.5-279.3grams
with the age range of three to five months were used in the experimental study. The
animals were bought from the animal house of faculty of natural sciences and were
49
fed with growers poultry feed in pelleted form and water. They were to stay for two
weeks for them to acclimatize.
3.3 CHEMICALS AND REAGENTS
Some of the chemicals and reagents used for this study were bought from scientific
shops in Ogbete market, Enugu and were of analytical grade. While others were
obtained from the laboratories of biochemistry departments caritas university and
Ebonyi state university. Some of them include: methanol, Alloxan and distilled water.
3.4 EQUIPMENTS AND APPARATUS
1. The equipments and apparatus used include:
2. Weighing balance
3. Glucometer
4. Test strips insulin injections
5. Needle and syringes
6. Disposable hand gloves
7. Pen and marker
8. Masking tape
9. Nose masks
50
10. Test tubes filter paper
11. Cotton wool
12. Soxhlet apparatus.
METHODOLOGY
3.5 PREPARATION OF PLANT MATERIAL
The fresh leaves of senna tora were extracted with 1,326 ml methanol to obtain the
extract in a soxhlet apparatus, then the was concentrated in a water bath at temperature
of 550 to obtain a paste like extract of the leaves.
3.6 WEIGHING AND GROUPING OF RATS.
The rats were weighed and grouped into four (4) groups of five rats each in separate
cages.
Group A: diabetic, administered with of
500mg/kg of Alloxan and 500mg/kg of
senna tora leaves extract.
Group B: diabetic, administered with of 500mg/kg of Alloxan and 1000mg/kg of
senna tora leaves extract.
Group C: served as diabetic control with only 500mg/kg of Alloxan.
Group D: served as a non diabetic control without Alloxan or the extract
3.7 INDUCTION OF DIABETES
51
Group A, B and C were rendered diabetic by injecting a freshly prepared aqueous
solution of Alloxan. The rats were injected with 100mg/kg body weight of alloxa
monohydrate each.
3.8 INJECTION OF SENNA TORA
Group A and B were injected with 500mg/kg body weight of senna tora respectively.
3.9 COLLECTION OF BLOOD SAMPLES
Blood samples were collected through the eye and tail veins. Samples were collected
as follows: before Alloxan induction, at 30 and 60 minutes after Alloxan induction
and at 30, 60, 90 and 120 minutes after senna tora injection.
3.10 BLOOD GLUCOSE ESTIMATION
Sample: whole blood
Kit used: one touch glucometer test kit.
3.11 PRINCIPLE: Glucose and oxygen react in the presence of gluconic acid and
hydrogen peroxide. In a reaction mediated by peroxidase producing a blue colour is
directly proportional to the glucose concentration in the sample.
52
3.12 PROCEDURE: The test strip was into the glucometer, the code written on the
strip container appeared and later disapproved. A drop of blood was spotted on the
sample spot and the blood glucose level result displayed on the meter within five
seconds in mg/dl.
3.12 NORMAL RANGES: the range of blood glucose level in human being is 70110mg/dl while that of albino rats is 80-95mg/dl
CHAPTER FOUR
4.0 RESULTS
The
blood
glucose estimation of the rats in different groups before Alloxan
induction, after Alloxan induction and also after senna tora induction are shown in
the tables below:
Table 1: Glucose level before Alloxan induction (mg/dl)
Group A
Group B
Group C
Group D
(Mg/dl)
(Mg/dl)
(Mg/dl)
(Mg/dl)
86
67
81
53
92
84
85
80
83
65
96
92
88
83
82
85
85
80
87
90
91
Table 2. Glucose level at 30 minutes after Alloxan induction
Group A
Group B
Group C
(mg/dl)
(mg/dl)
(mg/dl)
107
86
94
90
104
107
82
108
100
89
94
96
92
101
105
54
Table 3.Glucose level at 60 minutes (1 hr.) after Alloxan induction
Group A
Group B
Group C
(mg/dl)
(mg/dl)
(mg/dl)
118
99
120
110
141
123
97
158
120
101
132
119
112
121
126
Table 4.Glucose level at 30 minutes after senna tora injection
Group A
Group B
Group C (no injection)
(mg/dl)
(mg/dl)
(mg/dl)
115
80
118
105
130
131
94
153
116
97
128
117
102
109
122
55
Table 5.Glucose level at 60 minutes (1 hr.) after senna tora injection
Group A
Group B
Group C (no injection)
(mg/dl)
(mg/dl)
(mg/dl)
103
76
115
101
115
128
89
141
112
92
120
114
95
100
118
Table 6.Glucose level at 90 minutes (1½ hrs.) after senna tora injection
Group A
Group B
Group C (no injection)
(mg/dl)
(mg/dl)
(mg/dl)
95
65
111
92
93
122
78
119
106
89
100
109
90
91
101
56
Table 7.Glucose, level at 120 minutes (2 hrs.) after senna tora injection
Group A
Group B
Group
(mg/dl)
(mg/dl)
(mg/dl)
84
57
108
87
76
119
67
84
99
83
90
103
82
87
97
Table 8. Group D: Average glucose level without Alloxan and senna tora
No
of Before
rats
5
Alloxan
86
Before
Alloxan After senna tora injection
induction
30mins.
60 mins. 30 mins
60 mins. 90 mins. 120mins.
86.8
86.4
86.6
87
87.4
86
Table 9: % average increase of glucose level at one hour after Alloxan induction
Group A
Group B
Group C
57
26%
35.9%
29.6%
Table 10: % decrease of glucose at one after senna tora injection
Group A
Group B
12%
17.9%
HISTOGRAM REPRESENTATION OF THE RESULTS
140
120
100
80
60
40
20
0
1
2
3
4
585
6
7
8
9
Before
alloxan
inducation
84.3
30mins after
alloxan inducation
60mins (1hr) after
alloxan inducation
30mins after senna
tora inducation
1hr after senna
tora inducation
1hr 30mins after
senna tora
inducation
2hrs after senna tora
inducation
100mg/kg
100mg/kg
500/1000mg/kg
500/1000mg/kg
500/1000mg/kg
500/1000mg/kg
97
119.8
114.5
107.9
97.3
86.9
CHAPTER FIVE
5.0 DISCUSSION
Primary therapeutic purpose for treating type 1 diabetes is to reduce blood glucose
levels. Various hypoglycemic medications have been prescribed in the hospitals and
clinics which promote insulin sensitivity and reduce hepatic glucose output. However,
some of these hypoglycemic medications may have some effects. In case of type 1
diabetes patients who have been treated with insulin therapy for extended period of
time compound about pain, bruise and even insulin allergy rash and dyspnoea (Lee,
2008). Therefore natural medicinal plants and foods that have antidiabetic functions
but do not have harmful side effect have been focused on. Recently kinds of
59
polysaccharides from the edible plants have been reported as antidiabetic agents. In
the present study, positive effect of senna tora leaves extract on blood glucose control
was clearly seen in Alloxan diabetic rats. From the results obtained, there was an
increment in the random blood glucose levels at 30 and 60 minutes after the induction
of Alloxan however, the lessening of the blood glucose rise was seen even with a short
period of 30 minutes after injection of the rats with senna tora leaves extract. There
was a significant difference in the reduction of glucose levels amongst the different
groups. The reduction of blood glucose level is highest in group B than in other
groups followed by that in group A and finally in the group C. This might be related
with the concentration of senna tora leaves extract administered to the different
groups. Group B received 1000mg kg body weight of the extract while group C did
not receive any extract, however the three groups A, B and C received 100mg kg body
weight of Alloxan each before the administration of senna tora extract. Other possible
explanation in regards to the differences in reduction of blood glucose level is that
senna tora extract might protect some pancreatic cells from further damaging or even
enhancement of remaining beta cell function.
5.1 CONCLUSION
The result obtained showed that methanol extract of senna tora leaves has a beneficial
effect for hypoglycemic control, thus the extract can be used for the treatment of type
1 diabetes mellitus.
5.2 RECOMENDATION
60
Studies on the long-term effects of senna tora may be worthy to performed in the
future since this short term study has implications in terms of reducing blood glucose
levels and also further studies with pancreatic beta cells are needed to clarify its
effects, the mechanism involved and the Phytochemical properties.
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