RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

Annexure – II

PROFORMA FOR REGISTRATION OF SUBJECT FOR

DISSERTATION

1. / NAME OF THE CANDIDATE AND ADDRESS / Mr. NAGESH GOWDA. S
DEPARTMENT OF PHARMACOLOGY,
SET `s COLLEGE OF PHARMACY
S.R.NAGAR
DHARWAD 580002
2. / NAME OF THE INSTITUTION / SET `S COLLEGE OF PHARMACY,
S.R.NAGAR,
DHARWAD – 580002
3. / COURSE OF STUDY AND SUBJECT / MASTER OF PHARMACY IN
PHARMACOLOGY
4. / DATE OF ADDMISSION TO THE COURSE /
JUNE 2008
5. / TITLE OF THE TOPIC“EFFECT OF PIPER NIGRUM LINN. FRUIT EXTRACT ON TESTOSTERONE INDUCED BENIGN PROSTATIC HYPERPLASIA IN RATS”.
6.0 / BRIEF RESUME OF THE INTENDED WORK
6.1 Need for the study:
Benign prostate hyperplasia (BPH) is a benign and uncontrolled growth of prostate gland, which may lead to bladder out flow obstruction and lower urinary symptoms. It has been estimated that more than 50% of men older than 50 exhibit signs of BPH and approximately 45 million men will suffer from BPH by 20201. Between 15% to 30% of these men have lower urinary tract symptoms like smooth muscle dysfunction and urinary retention. In England and Wales up to 25% men undergo prostatectomy because of acute urinary retention, which doubles the risk of death & morbidity. More over, chronic urinary retention can lead to renal failure and is responsible for 15% prostatemies2.
The etiology of BPH, although not fully elucidated but involves hormonal changes in ageing man. The development and growth of the prostate glands depends on androgen stimulation, mainly by Dihydrotestosterone (DHT) which is formed in the prostate, as in the tissues, through the enzymatic conversion of testosterone into its more active metabolite DHT, catalyzed by prostate 5 apha- reductase. DHT binds to androgenic receptor and promotes protein synthesis and cellular growth, with ageing, production and accumulation of DHT in prostate increases, encouraging cell growth and causing hyperplasia
The pathogenesis of BPH still remains largely unresolved, but ageing, androgens3, inflammation4, and oxidative stress5 are the established risk factor for the development of BPH and benign prostatic enlargement (BPE). Therefore anti-androgenic drugs, mainly 5-alpha reductase inhibitors are indications for the treatment of BPH. BPH also involves augmented adrenergic tone in prostate smooth muscle, regulated through alpha-adrenoceptor. Therefore alpha-adrenoceptor blockers are also used in the treatment of BPH and are particularly effective for relaxing the smooth muscle and improving BPH symptoms2.
It is reported that many traditional plants like coconut oil 6, SPLE 7and permixon 8, are used in the treatment of BPH because these plants contains fatty acids like oleic acidand linoleic acid, which are responsible for 5 alpha-reductase activity, Plants which possesses anti-inflammatory9 and anti-oxidant properties10 are also useful in the treatment of BPH.
The Piper Nigrum Linn contains fatty acids like oleic acid, linoleic acid, alkaloids & volatile oils possesses anti-androgenic and anti-inflammatory activity11. The polyphenolic compound possesses anti-oxidant activity12.
Literature survey about Piper Nigrum Linn. Suggested the paucity of information regarding its effect on BPH. Therefore we have undertaken to investigate the effect of piper nigrum Linn. fruit extract on testosterone induced BPH in rats.
6.2 Review of literature
Piper nigrum Linn. (Black pepper) (Fam: piperaceae) is a stout globrous climbing perennial. Leaves are simple, alternate, cordate, very variable in breath. Flowers are minute in spikes, usually dioecious. Fruits are globular or oblong, externally blackish brown with raised reticulated wrinkles13, 14.
Piper nigrum contains fatty acids like oleic acid, linoleic acid, and palmitric acid11, alkaloids like piperine, piperamides, and guineensine. It also contains polyphenols and volatile oils like eugenol, safrol, myristicine14 etc.
In traditional system of medicine Piper nigrum is used as aphrodisiac, diuretic, urethrorrhea, arthritis, asthma, and dysentry13. It has been reported that fruit extract possesses anti-androgenic11, anti-oxidant12,15 properties and prevents inflammation16, lung carcinogenes17 and colon cancer18.
6.3 Objectives of study
The objective of the proposed study is to evaluate the:
Ø  Effect of alcoholic and petroleum ether extract of Piper nigrum Linn. fruits on prostate to body weight ratio
Ø  Effect of alcoholic and petroleum ether extract of Piper nigrum Linn. fruits on alpha adrenergic activity in rats using vasdeferens
Ø  Effect of alcoholic and petroleum ether extract of Piper nigrum Linn. fruits on histopathological changes of prostate
7. / MATERIALS AND METHODS
7.1 Source and collection of data:
The sources of data will be from the laboratory experiments, which involves extraction of the fruits and their pharmacological activities on experimental animals, such as male Wister rats. The result and data obtained from present study will be analyzed by one way analysis (ANOVA) test followed by Tukey`s test.
7.2 Method of collection of data:
Extraction: the dried fruit of Piper nigrum will be extracted exhaustively with ethanol and Pet.ether by using Soxhlet`s extractor.
Materials:
Ø  Chemicals: a) olive oil
b) Anesthetic ether (diethyl ether)
Ø  Standard drug: a) Phenylephrine
b) Testosterone
c) Finasteridae
Animals:
Male wistar rats weighing 150-200g will be used. The animals will be maintained under controlled condition of temperature (23 ± 20C), cycles. The animals are randomized into experimental and control groups and housed each in sanitized polypropylene cages containing sterile paddy husk.
1. In vivo study on the activity against experimentalprostatic Hyperplasia19. The male rats will be selected and randomized in to different groups.
Testosterone propionate (TP) (3mg/kg) in olive oil is administered by sc route for 21days to induce prostate hyperplasia. Finasteride (1mg/kg) is used as a standard, and olive oil is a vehicle and given alone to control groups of animals. Both ethanolic and Pet.ether extract of Piper nigrum fruit are administered orally for 21days along with TP (sc route).
On the 22nd day rats are been anesthetized using anesthetic ether. The prostate glands are collected and the prostate to body weight ratio will be recorded.
2. In vitro study on the alpha-adrenergic antagonistic activity19, 20.
The rats will be anesthetized with ether and the vasdeferens will be dissected and freed from extraneous tissue and suspended in organ bath containing Tyrode solution gassed with 95% O2 and 5% CO2 at 37o C. The contraction of the tissue will be recorded using phenylephrine in different concentration with or without Piper nigrum Linn. fruit extract.
3.  Histopathological studies:
After recording prostate to body weight ratio, the prostate are fixed in 10% neutral buffer formalin, and they are subjected to histological examination.
7.3 DOSE THE STUDY REQUIRES ANY INVESTIGATION OR INTERVENTION TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS? IF SO, PLEASE MENTION BRIEFLY.
Yes, the above study requires In Vivo screening techniques on male Wistar rats.
7.4 HAS ETHICAL CLEARANCE BEEN OBTAINED FROM INSTITUTION IN CASE OF?
The copy of the ethical clearance certificate obtained from institutional animal ethical committee (IAEC) is attached.

8.

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REFERENCES:

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2.  Thorpe A, Neal D. Benign prostatic hyperplasia. The Lancet 2003; 361(9366):1359-367.
3.  Vincenzo mirone, Ferdinando Fusco, Imbimbo. Androgens and benign prostatic hyperplasia. European urol 2006;5:410-17.
4.  Giacomo norvara, Antonio Galfano, Walter Artibani. Inflammation, apoptosis, and BPH: What is the Evidence? European Urol 2006; 5:401-09.
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9.  Zhang , L Liu , L Song . Effect of the polysaccharide fraction of urtica fissa on castrated rat prostate hyperplasia induced by testosterone propionate. Phytomedicine. (Article in press)
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11.  Noriko H, Masashi T. Testosterone 5 α-reductase inhibitory active constituents of piper nigrum leaf. Biol Pharm Bull 2007;30:2402-405.
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13.  Vaidyaratnam PSV. Indian Medicinal plants. Olient Longman Limited Mumbai 1995;4:297-02.
14.  Indian Herbal Pharmacopoeia. Indian Drug manufacturers Association Mumbai 2002;317-26.
15.  Gulcin I. The anti-oxidant and radical scavenging activities of black pepper seeds. International J of biochem 2008; 12: 94-99.
16.  Mujumdar AM, Dhuley JN. Anti-inflammatory activity of piperine. Japan J Med Sci Biol.1990; 43(3):95-100.
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20.  Khakha BS, Surperenant A, Humphrey PPA. A study on P2X purinoreceptor mediating the electrophysiological and contractile effect on purine nucleotide in rat vasdeference. British J pharmacol 1985;115: 177-85

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