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Transcript
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA-BANGALORE
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR
DISSERATATION
1.
NAME OF THE CANDIDATE
ASHOK A MUCHANDI
DEPARTMENT OF PHARMACOLOGY
H.S.K COLLEGE OF PHARMACY
B.V.V.S.CAMPUS,
BAGALKOT-587101
2.
NAME OF THE INSTITUTION
H.S.K COLLEGE OF PHARMACY
B.V.V.S.CAMPUS, BAGALKOT-587101
3.
COURSE OF STUDY AND SUBJECT
4.
DATE OF ADMISSION TO COURSE
5.
TITLE OF THE TOPIC:
MASTER OF PHARMACY IN PHARMACOLOGY
“Hepatoprotective activity of Stereospermum suaveolens DC on Albino rats.”
1
6.
BRIEF RESUME OF THE INTENDED WORK
6.1 Need for the study:
Free radical reactions have been implicated in the pathology of many human diseases
including atherosclerosis, myocardial infraction, diabetics, neurodegenerative conditions, cirrhosis and
other disease conditions. Radicals and other reactive species are formed constantly in human body,
are removed by enzymatic and non-enzymatic antioxidative defense system. Oxidative stress
occurring when antioxidant defenses or inadequate can damages lipids, proteins, carbohydrates and
DNA1
Liver the key organs of metabolism and excretion are constantly endowed with the task
detoxification of xenobiotics, environmental pollutants, contaminations of water like carbon tetra
chloride, fluoride and other metal2. Fluoride is an environmental and industrial pollutants is already
has been reported that, accumulation of fluoride alters activities of some enzymes involved in
antioxidant defense system3. While a curative agent has not yet been found in modern medicine. A
current usage of corticosteroids and immune suppressive agents are only brought about symptomatic
relief. Further more their usage is associated with relapse and danger of side effects4
Management of liver diseases is still a challenging to the modern medicine. Various herbal
and hepatomineral preparation are extensively used for the various disorders. Many formulations are
available in for treating liver disorders like LIV –52, HD-03 5 and Peumus boldus. Ocimum sanctum6.
Hence in the ethnic medicines have safe and have multitherapeutic applications. With this objectives,
traditionally and in folk medicine claim Stereospermum suaveolens DC were selected for
hepatoprotective activity in albino rats.
2
6.2 Review of literature
Drug Profile:
Title of plant : Stereospermum suaveolens DC
Family
: Bignoniaceae , Bignonia-flowers looks like trumpet
Synonyms
: Patala, Paral, Padal, Kalagora, Padari
Parts used
: Root bark
Habitat
:
In India-Bihar, Bengal, Terrain region of Himalaya and Burma.
Chemical constituents: Root fat contains- Palmitic (30.41%), Stearic (58.16%), Oleic acids (11.43%)
and Ceryl alcohol, Extract of plant contins-Lapachol, Dinatin, Dianatin-7-glucuroniside1, and βsitosterol. It also contains saponin, α- cellulose, lignin, non-drying fatty oil and mucilage7.
Traditional uses: Externally it is used as analgesic, wound healing property. Internal uses are mainly
antidyspeptic, astringent, and liver stimulant. Also useful in vomiting, diarrhea, in asthma and as
general tonic. Flowers are used in semen debility8.
Pharmacological actions: Scientifically validated that this plant shows hypoglycemic activity9,
anticancer activity against human epidermoid carcinoma of nasopharynx in tissue culture10.
6.3 Objectives of the study: The plant is collected and authenticated by botany staff, Ayurvedic
college of this campus. The extract will obtain by using non-polar solvents and polar solvents. The
dose of drug extract will be selected on earlier study.
1. In vivo evaluation plant extract of CCl4 induced hepatotoxicity in rats.
2. In vivo evaluation plant extract on NaF induced oxidative stress.
3
7.
METHODS AND MATERIALS
7.1 Source of data:
Data will be collected from experimental animals using standard parameters and selection of doses
is based on earlier study.
Experimental animal groups:
CCl4 INDUCED HEPATOTOXICITY IN RATS
a) Group I
- Control/positive group, receives only normal saline, (n= 8)
b) Group II - Negative control receives CCl4,( n=8)
c) Group III - Effect of standard drug-Liv 52 on CCl4 induced hepatotoxicity in rats, (n=8)
d) Group IV - Effect of low dose of extract on CCl4 induced hepatotoxicity in rats, (n=8)
e) Group V - Effect of moderate dose of extract on CCl4 induced hepatotoxicity in rats, (n=8)
f) Group VI - Effect of high dose of extract on CCl4 induced hepatotoxicity in rats, (n=8)
NaF INDUCED OXIDESSIVE STRESS IN MICE
a) Group I -Control/positive group, receives only normal saline, (n= 8)
b) Group II -Negative control receives NaF, ( n=8)
c) Group III-Effect of standard drug Liv- 52 on NaF induced oxidative stress in mice, (n=8)
d) Group IV-Effect of low dose of extract on NaF induced oxidative stress in mice, (n=8)
e) Group V -Effect of moderate dose of extract on NaF induced oxidative stress in mice,(n=8)
f) Group VI -Effect of high dose of extract on NaF induced oxidative stress in mice, (n=8)
4
7.2 Materials:
Drug
: Root extract of Stereospermum suaveolens DC.
Animals
: Wister albino rats of either sex and Swiss albino mice of either sex
Instruments: Tissue homogenizer, centrifuge, enzyme assay kits and autoanalyser etc
Chemicals : All the chemicals to be used in the present study will be AR graded
7.3 Methods:
Preparation of plant extract-The roots are authenticated and collected in ideal conditions, will be air
dried, saved and powdered to a fine powder and passed through a sieve no 44.The collected powder
will be extracted using polar and non polar solvents. The present study will be carried out by
following two experimental models
CCl4 induced hepatotoxicity- Rats of various groups containing 8 animals. Each group was divided
as rats of group I, served as normal; group II, served as CCl4 treated group. Rats of group I and II
received the vehicle. Rats of groups IV, V and VI are receives low, moderate and high doses of
extract for a period of 19 days. On days 15th, 17th and 19th of the rats of groups IV, V and VI receives
1 ml / kg body weight of CCl4 in liquid paraffin (1:1) orally and I groups are not given CCl4. 24 hrs
after the last dose of CCl4, blood was collected by retro-orbital puncture and liver was perfused with
normal saline and stored in cold condition until assay11.
NaF induced oxidative stress- To determine the dose dependent activity of extract for hepatic
protection, four different groups of animals – Group I, receives normal saline and Group II receives
NaF treated group. Group IV, V and VI treated with different doses of extract (low, moderate and high
according to body weight) for 10 days prior to NaF treatment through their drinking water (7 days at
the dose of 600 ppm). After 24 hrs the last dose of NaF administration, all the mice were sacrificed.
5
Liver tissues are homogenates for all experimental animals was collected and estimated for enzyme
assay12.
Evaluation of biochemical parameters- The parameters are estimated for serum glutamate
oxalloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), glutathione,
superoxide dismutase (SOD), catalase and lipid peroxidation in liver homogenate by using
spectrophotometric method. Total protein content will estimate by Lowry method.13-14
Histopathological studies- Two animals from each group were scarified on the day of blood
withdrawal and enzyme assays. Liver is isolated. Tissue is immersed in 10% formalin for
histopathological studies15.
7.4.
Dose the study require any investigations or interventions to be conducted on patients
or other humans/animals? If so please
describe briefly
Yes, for the study rats and mice rats will be used. The effects of preparation on liver will be studied by
considering physiological, pathological, biochemical parameters using animal models.
7.5 Has ethical clearance been obtained from your institution for performing various tests on
animals?
Yes, the study is cleared from Institutional animal’s ethics committee and the copy is enclosed with
the protocol.
6
8.
REFERENCES:
1. Mitra, S.K., Venkataranganna, M.V., Sundaram, R. and Gopumadhavan, S. Antioxidant Activity of
AO-8, Herbal Formulation in vitro and in vivo experimental models. Phyt. Res., 1999, 13:300-303.
2. Butler, T.C. Reduction of CCl4 in vivo and reduction of CCl4 and CHCl3 in vitro by tissues and
tissue constituents. J. Pharm. Exp. Ther., 1961, 134:311-318.
3. Patel, P.D., Chinoy, N.J. Influence of Fluoride on biological free radical reactions in ovary of mice
and its reversal. 1998. Fluoride 31., 3:S27.
4. Handa, S.S., Sharma, A. Hepatoprotective activity of Andrographolide against Galactosamine and
paracetamol intoxification in rats. Ind. J. of Med. Res., 1990, 92:284-292.
5. Mitra, S.K., Venkataranganna, M.V., Sundaram, R., Gopumadhavan, S. Protective effect of HD-03,
a herbal formulation, against various hepatotoxic agents in rats. J. of Ethn., 1998, 63:181-186.
6. Chattopadhyay, R.R., Sarkar, S.S., Ganguly, S. Hepatoprotective activity of Ocimum sanctum leaf
extract against paracetamol induced Hepatic damage in rats. Ind. J. of Pharm., 1992, 24:163-165.
7. Chattarjee, Asma., Chandra, Pakrashi, Satyesh. The treatise on Indian medicinal plants. National
Institute of Science Communication, New Delhi, 2000. 2:10-11.
8. Chattarjee, Asma., Chandra, Pakrashi, Satyesh. The treatise on Indian medicinal plants. National
institute of science communication, New Delhi, 1997. 5:46-47
9. Sastry, J.L.N., Illustrated Dravyaguna Vijnana, 5th ed. Choukhambha Orientation, Varanasi, 2005,
2:403.
10. Ayurveda. The wealth of India. New Delhi council of Scientific and Industrial Research, 2003,
vol X Sp-w:51-52.
11. Mitra, S.K., Venkataranganna, M.V., Sundaram, R. and Gopumadhavan. Effect of HD-03, a herbal
formulation, on the Antioxidant Defense System in Rats. Phyt. Res., 1998, 12:114-117.
7
12. Sinha, M., Manna, P., Sil, P. Aqueous extract of the bark of Terminalia arjuna plays a protective
role against sodium fluoride induced hepatic and renal oxidative stress. J. Nat. Med., 2007, 61:251260
13. Reitman, S., Frankel, S., A colorimetric method for the determination of serum glutamic
oxaloacetic and glutamic pyruvic transaminases. Americ. J. Clin. Path., 1957, 28:56-63.
14. Lowry, O. H., Rosebrough, N.J., Farr, A.L., Randall, R.J. Protein measurement with the FolinPhenol reagent. J. Bio. Chem., 1951, 193: 265-272.
15. Cameron, G.R., and Karunarathe, W. A. E. Carbon tetrachloride cirrhosis in relation to liver
degeneration. J. Path. Bacteriol., 1936, 42:1.
8
9.
SIGNATURE OF CANDIDATE
( Mr. ASHOK A MUCHANDI )
10.
REMARKS OF THE GUIDE
This detail literature survey indicates that, the
level of enzymes SGPT, SGOT, SOD and liver
necrosis and fatty liver in the animal are
important factors for evaluating the
hepatoprotective activity of extract. Most of the
plant medicines reported hepatoprotective and
neuroprotective. On this basis, this work has been
selected for evaluation.
11.
NAME AND DESIGNATION OF
THE GUIDE
Mr. V.M.CHANDRASHEKHAR
Assistant Professor
12.
SIGNATURE
13.
CO-GUIDE
--------------------
14.
SIGNATURE
-------------------
15.
HEAD OF THE DEPARTMENT
16.
SIGNATURE
Prof. I.S.MUCHANDI
Department of Pharmacology
H.S.K College of Pharmacy
BAGALKOT- 587101
9
17.
REMARKS OF THE PRINCIPAL
The above mentioned information is correct and
I recommend the same for approval.
18.
NAME OF THE PRINCIPAL
19.
SIGNATURE
Prof .I .S. MUCHANDI
Principal and H.O.D.,
Department of Pharmacology
H.S.K. College of Pharmacy
BAGALKOT-587101
10
OFFICE OF THE INSTITUTIONAL ANIMAL ETHICS
COMMITTEE (IAEC)
HANAGAL NSHRI KUMARESHWAR COLLEGE OF PHARMACY,
BAGALKOT-587101, KARNATAKA
REG NO.821/01/a/CPCSEA, Dated: 6th AUG 2004 UNDER THE RULES 5(a) OF THE
“ BREEDING OF AND EXPERIMENTS ON ANIMALS (Control and Supervision)
Rules 1998”
Ref: HSKCP/IAEC, Clear / 2007-08 / 1-8
Date:
CERTIFICATE
This is to certify that Mr. ASHOK A MUCHANDI a student of FIRST
M.Pharm is permitted to carry out experiments on animals for the dissertation /
thesis work entitled ‘Hepatoprotective activity of Stereospermum suaveolens DC. on
albino rats’ as per details mentioned and after observing the usual formalities
laid down by IAEC as per provision made by CPCSEA.
S
Animal house in charge
CHAIRMAN
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