Annexure – II proforma for registration of subjects for dissertation




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Rajiv Gandhi University of Health Sciences, Karnataka, Bangalore

Annexure – II




PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION





01

Name and Address of the Candidate

THIPPESWAMY.B

S/o Gangadharappa.B

Doulathpur(P)

Sandur(T)

Bellary(D),KARNATAKA

02

Name of the Institution

T. M. A. E. Society’s

S. C. S. College of Pharmacy,

Harapanahalli – 583 131

KARNATAKA

03

Course of the Study

Branch


M.Pharm. (Pharmacology)

04

Date of Admission to course

01.06.2007

05

Title of the Topic

A Study on antioxidant and organ protective effect of aerial part of

Sonchus asper(L.) Hill.

06

Brief resume of the intended work


6.1. Need for the Study



Enclosure – I





6.2. Review of the Literature

Enclosure – II

6.3. Objective of the Study


Enclosure – III

07

Materials and Methods


7.1. Source of data



Enclosure – IV

7.2. Methods of collection of data


Enclosure – V

7.3. Does the study require any

Investigations on animals?

If yes give details


Enclosure – VI

7.4. Has ethical clearance been

obtained form your institution

in case of 7.3.


Yes, Registration No: 157 / 1999/ CPCSEA

(Copies enclosed)

08

List of References


Enclosure – VII


ENCLOSURE-I



06. Brief resume of Intended Work:

6.1 Need For The Study:

Human beings are exposed to various stress conditions due to the environmental pollution, modern life style, xenobiotics, etc. During such conditions many free radicals are being produced within the body and may react with tissue membrane macro-molecules cause lipid peroxidation and tissue necrosis. However, there are several inbuilt protective mechanisms like tissue glutathione (GSH), glutathione dismutase, etc.

Continual exposure to such stressful conditions generates free radicals, which may over power the inbuilt protective mechanisms and cause tissue damage. Search for remedies/drugs for treating the diseases/organ damage with minimal side effects is in force since ancient periods. Herbs/herbal products are found to be highly effective in this regard. In our search for herbal remedies, we found a plant by name Sonchus asper (L.) Hill. Upon literature review, it was found that the plant possesses flavonoids1 and other polyphenolic compounds4, which are known free radical scavengers. Similarly there was no report in the literature regarding its phytochemical, antioxidant and organ protective effects, hence, it has been planned to take the aerial part of this plant for the study to evaluate the antioxidant and organo protective effects. Since, this plant is grown widely and available at hand stretch; the study may be useful in exploring one more herb for the purpose. In addition it may further reduce the cost of treatment or prevention of organ disorders. Therefore the study is needed and justifiable.


ENCLOSURE-II




6.2 Review of Literature:
The literature survey reveals that the plant Sonchus asper (L.) Hill belongs to family Asteraceae or compositae with the synonyms Lyon sheart, Spiny sow thistle, Prickly sow thistle have been reported to contain flavonoids1( quercetin 3-o-glucoside, luteolin 7-o-glucoside etc). The phyto-chemical profile of this plant reveals the presence of α-amyrin (4.0), β-amyrin (8.0), germanicol (6.0), taraxasterol (17.0), lupeol2 (28%), epifriedelinol acetate, stigmasterol, apigenin and luteolin3. There are reported that the tender leaves of Sonchus asper (L.) Hill.contains polyphenols4 (108.3). The plant can be used as an anti-oxidant & for the treatment of diabetes and cancer5. The local folklore practitioner suggested that the plant possess hepatoprotective, anti-inflammatory, wound healing properties, anti-ulcer activities. The plant has protective effect on cardiac abnormalities in experimental hyper-cholesterolemia and also has good cardiac protective effects6.

At Loralai, the plant is pounded and applied to wounds or boils7. In Spain; it is commonly used as an emolient7. The stems have been peeled and eaten raw like celery8. In the literature, there are no clear cut reports on the organ protective (antioxidant, hepatoprotective, gastroprotective and nephroprotective) activity of this plant and even the phytochemical properties of this plant are incomplete, hence, the present study is designed to explore the valuable pharmacological activities of the aerial part of the plant Sonchus asper (L.) Hill.



ENCLOSURE – III




6.3 Objectives of the study:
Since there is incomplete phytochemical and pharmacological profile, it is planned to undertake a study on the bark of this plant with the following objectives:


  1. To prepare various extracts (petroleum ether extract, chloroform extract, alcoholic extract 70%, aqueous extract) by successive extraction technique of the aerial part of Sonchus asper (L.) Hill. We have planned to use70% ethanolic extract of the plant for various pharmacological activities.

  2. To identify pharmacological profile of the aerial part of the plant:

  1. To assess antioxidant property.

  2. To assess acute toxicity of the aerial part of Sonchus asper (L.) Hill.

  3. To assess organ protective activities on aerial part against experimentally induced hepatotoxicity (CCl4, Paracetamol and Thiocetamide induced hepatotoxicity in rats), nephroprotective activity against paracetamol, cisplatin and gentamicin induced nephro-toxicity in rats and gastro protective activity against experimentally induced gastric ulceration (Pyloric ligation, Aspirin and Alcohol induced ulcer in rats).


ENCLOSURE – IV



7. Material & methods:

7.1 Source of data:

Whole work is planned to generate data from laboratory i.e., experiments on animals. The rat and mice will be used for this purpose. Standard analytical procedures will be adopted for estimation of biochemical like blood creatinine, blood urea and other biochemical like SGPT, SGOT, ALP, bilirubin (Total and Direct), blood urea, blood creatinine etc. Some in-vitro and in-vivo studies like antioxidant property is also planned to generate the data. It is also planned to use the available literature for interpreting the data.




ENCLOSURE – V

The whole study is divided into four phases to generate data.

Following are the four phases:

Phase – I: It is planned to collect the plant material, prepare the extracts (petroleum ether extract, chloroform extract, alcoholic extract and aqueous extract) and qualitatively analyze them for the presence of the types of phytoconstituents of aerial part of Sonchus asper (L.) Hill. In this phase 70% ethanolic extraction is used for further investigation

Phase – II: In this phase, it is intended to establish the correlation between antioxidant and organ protective activity against experimentally induced hepatotoxicity10,11 (CCl4, Paracetamol and Thiocetamide induced hepatotoxicity in rats).

For antioxidant activity we are using the following models:



  1. In vitro models

  1. Reducing power(Oyaizu, 1986)

  2. Superoxide anion scavenging activity(Nishimki et al., 1972)

  3. Hydroxyl radical scavenging activity(Halliwell and Gutteridge)

  4. Nitric oxide radical scavenging activity




  1. In vivo models

1)Glutathione estimation(Aykae, et al.)

2)Lipid peroxidation(John Buege A et al.)



Phase – III: This phase is designed so as to utilize various pharmacological tools as well as to analyze blood and tissue level of vital biochemical parameters like SGOT, SGPT, ALP, bilirubin (total & direct), creatinine, blood urea, etc..
Phase – IV: To assess gastro protective activity against experimentally induced gastric ulceration (Pyloric ligation, Aspirin/Indomethacin and Alcohol induced ulcer in rats), all these models are used to generate the data to support the need for the study.

The Study Design, Criteria and Plan of Work Are Outlined As Below:--

  • Inclusion Criteria for the selection of animals:--

Sex: - Both sex Age: Adult animals

Weight: - 150 – 240 grams Health condition: - Healthy



  • Exclusion criteria:-Any animal not confirming with above criteria are not selected for the experiment.

  • Study sampling: each model of organ toxicity requires five groups of six animals each.

Since the animal models of toxicity are mortal, there is no need to undergo follow up observation.

  • Parameters of study: In hepatotoxicity study, biochemical parameters like SGOT, SGPT. ALP, bilirubin(Total & direct), creatinine, blood urea etc. will be assessed, compared with positive control group & subjected to ANOVA test for statistical evaluation. In addition histopathological assessment will also be done as a part of this investigation.

  • Duration of study: Ten months


ENCLOSURE – VI



7.3 The proposed study requires investigation on albino mice for determination of LD50.

Albino rats for organ protective activities

Experiments are to be conducted on anaesthetized rats.

ENCLOSURE – VII


8.0 List of references:

01. Bruce A.Bohm, Tod F. Stuessy. Flavonoids of the Sunflower Family(Asteraceae) 2001:535

02. Ram.P. Rastogi, Mehrotra. Compendium of Indian Med. Plants.1993;(3):600

03. Ram.P. Rastogi, Mehrotra. Compendium of Indian Med. Plants.1991;(2):637

04. The Local Food-Nutraceuticals Consortium. Understanding local Mediterranean

diets: A multidisciplinary pharmacological and ethnobotanical approach,

Pharmacological Research. June 2005;1(52): 353-366.

05. Meenakshi Dhanawat,Gajendra Kamal Singh, Arindam Paul. Pharmacology and potential therapeutic uses of quercetin-a plant flavonoid, Indian Journal of Natural Products. June2005;2(21):3-11

06. Varatharajan Sudhahar, Sekar Ashok Kumar et al. Protective effect of lupeol and its ester on cardiac abnormalities in experimental hypercholesterolemia, Vascular Pharmacology2007;6 (46): 412-418.

07. Kirtikar KR, Basu BD. Indian Medicinal Plants.1999;(2):1441-1445.

08. Moerman.D. Native American Ethnobotany Timber Press. Oregon. 1998 ISBN

0-88192-453-9

09. Maria Mamani-Matsuda, Jerome Rambert, Denis Malvy et al. Quercetin Induces Apoptosis of Trypanosoma brucei gambiense and Decreases the Proinflammatory Response of Human Macrophages, Antimicrobial agents and Chemotherapy. March2004; 48(3): 924–929.

10. Shivakumar SI, Suresh HM, Kumar GS, Hepatoprotective activity of fruits of coccina grandis against CCl4 induced hepatotoxicity, Adv.Pharmcol Toxicol, 2006;7(1):7-9



11. Suja SR, Latha PG, Assessment of hepatoprotective and free radical scavenging effect of Rhinacanthus nausta(Linn) Kurtz in rats, Journal of, Journal of natural Remedies 2004;4(1): 66-72



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