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“PHARMACOLOGICAL EVALUATION OF ETHANOLIC LEAF EXTRACT OF
DALBERGIA SISSOO FOR ITS ANTIDIABETIC ACTIVITY”
BRIEF RESUME OF THE INTENDED WORK:6.1 NEED FOR THE STUDY:
The present epidemiological data revealed that diabetes is likely to remain a significant threat
to public health in the years to come. In the absence of effective and affordable interventions
for both types of diabetes, the frequency of the disease will escalate worldwide, with a major
impact on the population of developing countries1. The World Health Organization (WHO)
has predicted that the world wide number of patients with diabetes will double by the year
2025, from the current number of approximately 150 million to 300 million2. Despite the
considerable progress in the understanding and management of diabetes, the disease and the
complications related to it continue to increase3.
Diabetes mellitus is a heterogeneous metabolic disorder as old as mankind and
its incidence is considered to be high (4-5%) all over the world4. Diabetes is a
chronic disorder of carbohydrate, fat and protein metabolism characterized by
elevation of both fasting and post-paradinal blood sugar levels. This disorder is
characterized by chronic hyperglycemia and abnormality of lipid profile such as cholesterol,
low and high density lipoprotein and triglyceride leading to series of secondary complications
(5,6)
. The synthetic oral hypoglycemic agents can produce serious side effects in addition they
are not considered safe for use during pregnancy (7,8).
Medicinal plants have played a major role therapeutic agent. A multitude of herbs, spices
and other plant materials have been described for the treatment of diabetes
throughout the world. The medicinal plant acts as a useful source of new oral hypoglycemic
compounds for development of pharmaceutical entities. Few of the plants used for the
treatment of diabetes have received scientific or medical scrutiny and committee on
diabetes recommends that this area warrant further attention (9,10).
Despite the presence of known antidiabetic medicines in the pharmaceutical
market,
screening for new antidiabetic sources from natural plants is becoming more important
so that we can have alternative and safe effect on diabetes mellitus.
Attempts will be made to study and evaluate the anti-diabetic activity of the leaf extract
of Dalbergia sissoo. Shisham (Dalbergia sissoo Roxb), a deciduous tree of family
Fabaceae, is an important plant of great economic importance. Shisham wood is used in
furniture, construction work, agricultural implements, plywood industries and fuel purposes.
According to Greeks, Shisham plant has some medicinal properties. It provides financial
support to the farmers as it is considered as cash plant. Dalbergia sissoo Roxb. (Fabaceae),
known as Indian Rosewood, is reported to be useful in many conditions including fever,
ulcers, digestive disorders, and skin diseases
(11,12)
. Dried leaves of Dalbergia sissoo is
reported to have antibacterial, anti protozoan and anti inflammatory activity
(13)
. By
keeping in view of the importance of Dalbergia sissoo plant, the present study was designed
to evaluate the antidiabetic potential of ethanolic extract of Dalbergia sissoo in streptozotocin
induced diabetic rats.
6.2 REVIEW OF LITERATURE
In ayurveda the leaf juice of Dalbergia sissoo is used for eye ailments, The wood and
bark of Dalbergia sissoo
acts as abortifacient, anthelmintic, antipyretic, apertif,
aphrodiasiac, expectorant and refrigerant (10).
Ethanolic extract of the fruits of Dalbergia sissoo exhibited molluscicide effect against
eggs of the freshwater snail Biomphalaria pfeifferi(14).
The active extract of Dalbergia sissoo bark contains carbohydrates, phenolic compounds,
flavonoids and tannins. The ethanolic extract of the bark at a dose of 1000 mg/kg had the
most potent anti-inflammatory activity (15).
The leaves and trunk exudates of Dalbergia sissoo contain various compounds like
dalbergenone, dalbergin and methyl dalbergin, 4-phenyl chromene, dalbergichromene
(16, 17)
.
The plant also contains dalbergichromene, nordalbergin and isodalbergin as minor
constituents (18-22).
6.3 OBJECTIVES OF THE STUDY

Collection, Identification and authentication of the plant material.

Extraction of the plant material with solvents using direct soxhlet method.

Phytochemical analysis of the extracts.

Invivo pharmacological evaluation of the extract for it antidiabetic activity
b) MATERIALS AND METHODS
7.1 SOURCE OF DATA:
Collection & Authentication of Plant Material:
Dalbergia sissoo leaves will be collected from Acharya N.G Ranga University, Hyderabad,
India and will be authenticated by taxonomist.
Experimental Animals:
Sprague Dawley rats of both sexes weighing (200 to 300 g) will be maintained under
controlled conditions of (12 h Light and Dark Cycles) and temperature 25°C ± 1°C in the
Animal House, for two weeks prior to the experiment for acclimatisation. Animals had access
to food and water ad libitum. All pharmacological activities will be carried out as per
CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on
Animals) norms after obtaining the approval from the Institutional Animal Ethical
Committee.

The literature survey will be done by referring the abstracts and articles of all the
National and International Journals of Pharmacology.

National Research laboratories like IICT CCMB, NIN.

Science Direct, PubMed and other online E journals.
7.2. Method:
Ethanolic extracts will be prepared by steeping 40 grams of powdered plant material in 90%
ethanol for 48 hrs, using soxhlet extraction method followed by filtration. The extracts will be
concentrated to dryness by evaporation, under reduced pressure, in a rotary evaporator
(Heidolf,Germany) at temperatures below 45 °C. Stock solutions of the extracts will be made
fresh, by dissolving 4 g of dry extract in 20 ml dimethylsulfoxide (DMSO) and then makeup
to 100 ml volume with dechlorinated tap water.
Phytochemical Evaluation:
Qualitative phytochemical analysis of the ethanolic extract of Dalbergia sissoo will be
carried out using standard procedures to assess the different types of phytochemical
constituents present in the leaves of D. sissoo using different chemical tests. Screenings will
be done for carbohydrates, glycosides, proteins, amino acids, phytosterols, saponins,
flavonoids, alkaloids and tannins (23-25).
Induction of Diabetes:
To attain the hyperglycemic state rats, a single intraperitoneal injection of
Streptozoticin (STZ) 32 mg/kg body wt. dissolved in 0.1 M citrate buffer, pH 4.5.
Streptozoticin solutions will be prepared freshly due to limited stability of the compound.
After 72 hours, diabetes will be confirmed by the determination of fasting blood glucose.
Rats with blood glucose levels <180 mg/dL were excluded from the study and rest will be
distributed into different groups. Rats were kept in separate cages (one per cage) due to
excess urination in STZ induced rats. The bed of the cage will be monitored and if necessary
they will be changed every day. Body weight and fasting blood glucose levels of all the rats
will be measured before and at different weeks after the initiation of the experiment.
Studies of Acute Toxicity:
Acute toxicity studies will be performed on SD rats according to standard procedures.
Ethanolic extracts at doses of 50, 100, 300, 1000, and 3000 mg/kg body weight will be
administered to separate groups of rats (n = 5) after overnight fasting. Subsequent to
administration of drug extract, the animals are observed closely for the first 3 hours for any
toxic manifestations. Subsequent observations will be made at regular intervals for 24 h. The
animals will be further kept for observation for a week (26).
Pharmacological Evaluations of the extract:
1) Estimation of Blood glucose levels.
2) Estimation of Body weights, daily food and water intake.
3) Antioxidant capacity of ethanolic extract
a. DPPH radical scavenging activity
b. H2O2 scavenging activity (FOX assay)
4) Following Biochemical Estimations will be carried out after 28 days of sub-acute
toxicity studies.
1) SGPT
2) SGOT
3) ALP
4) ACP
5) Antioxidants Enzymatic Assays
a. Superoxide dismutase (SOD)
b. Catalase (CAT)
c. Reduced Glutathione (GSH)
d. Glutathione Peroxidase (GPx)
e. Glutathione S-transferase (GST)
6) Organ Weights will be taken after the duration of exposure
7) Histopathological studies will be carried out on vital organs
STATISTICAL ANALYSIS:
The results will be analyzed by using one-way ANOVA to establish the statistical
significance. The least significant difference will be used to determine significant differences
between individual samples. Values with p < 0.05 will be considered as significant.
7.3 Does the study require any investigations or invention to be conducted on patients or
other human or animals? If so, please mention briefly.
YES
Study requires investigation on animals. The effects of drug will be studied on various
parameters using rats as experimental animals.
7.4
Has ethical clearance been obtained from your institution in
case of 7.3?
Ethical committee approval letter is enclosed.
c) REFERENCES
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