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6. 1 Need for the study


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BRIEF RESUME OF INTENDED WORK:
6.1 Need for the study:
Peptic ulcers are the areas of degeneration and necrosis of gastrointestinal mucosa exposed to acid peptic secretions. Though they can occur at any level of the alimentary tract that is exposed to hydrochloric acid pepsin, they occur most commonly (98­­-99%) in either the duodenum or the stomach in the ratio of 4:1. Peptic ulcers are common in the present -day life of the industrialised and civilised world. The immediate cause of peptic ulcer disease is disturbance in normal protective mucosal barrier by acid - pepsin, resulting in digestion of the mucosa.1 It results probably due to an imbalance between the aggressive (acid and pepsin) and the defensive (gastric mucus and bicarbonate secretion, innate resistance of the mucosal cells) factors. A variety of psychosomatic, humoral and vascular derangements have been implicated.2

Even though a range of drugs available for the treatment of ulcers, many of these do not fulfill all the requirements and side effects such as arrhythmias, impotence and haematopoietic changes are noted. There are many agents in alternative medicine, which have shown promising antiulcer activity without producing above -mentioned adverse reactions.3

The literature survey reveals that Mentha arvensis Linn. leaves have been used as stimulant, antispasmodic, nervine, antiemetic, carminative and antihysteric properties.4,7 The leaves are acrid , aromatic, thermogenic, anodyne, deodorant, antiseptic, vulnerary, anthelmintic, digestive, stomachic, cardiotonic, expectorant, diuretic, depurative, sudorific, emmenagogue, dentifrice, febrifuge, contraceptive, wounds, cuts, helminthiasis, vomiting, diarrhoea, cardiac debility, cough, asthma, bronchitis, skin diseases, jaundice, cephalagia and general weakness,5 but no scientific and methodological investigations have so far been reported in literature regarding its action on gastric ulcers.

Hence the present study has been designed to evaluate the gastric antisecretory and antiulcer property of leaves of Mentha arvensis Linn. in albino rats.







6.2 REVIEW OF LITERATURE :


Mentha arvensis Linn.(Lamiaceae) commonly known as pudina. Leaves narrowed below, stalked, ovate and growing throughout India.6 Phytochemical rewiew reveals that the major constituents of leaves is essential oil : carvone, flavonoids, L –limonene, dihydrocarvone, carvomenthone, iso-menthone, dihydrocarveylacetate, diometin-7-glucoside, diosmin, diosmetin-7-0-beta-D-glucuronide, and luteolin-3-beta-D-glucuronide.7

The essential oil also contains menthol, menthyl acetate, menthone, piperitone, furfural, camphene, caryophyllene, dl-sesquiterpene alcohol, phenyl acetate, and other free and esterified fatty acids.7 and also contains tannins, lactones, and carbohydrates.17


Traditionally, it is employed for the treatment of jaundice, hepatopathy, asthma, cough, diarrhoea, bronchitis, fever, cephalagia, and general weakness.5

Review of literature shows Mentha arvensis Linn. leaves shows antioxidant activity,8, 9,nitric oxide scavenging activity,10 inhibition of immunologic and non- immunologic stimulation.11 It also possess anticandida,13 antifertility,14,23 antibacterial,16,19 antidiarrhoeal,17spermicidal,20 contraceptive,21 organogenesis,15 nitrification inhibitor,18 and fructolysis effect.22 Review of literature shows Menthol, 8 caryophyllene7 and flavonoids is known to possess a potent antiulcer activity. Mentha arvensis Linn. As an antiulcer is not reported so far. The above facts of folklore use as well as review of literature will have positive co-relation in relation to antiulcer property of Mentha arvensis Linn. Therefore, the present work has been designed to investigate the ethanol extract of Mentha arvensis Linn. Leaves on gastric antisecretory and antiulcer property in albino rats.


6.3 OBJECTIVE OF THE STUDY:


  • To assess gastric antisecretary and antiulcer activity of Mentha arvensis Linn. by experimentally inducing ulcer models in rats

  • To evaluate the possible mechanism of action of Mentha arvensis Linn. by measuring the various biochemical parameters in gastric ulcer.

  • To assess the cytoprotective property of Mentha arvensis Linn. by histo pathological investigations in ethanol induced ulcer model.





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METHODS AND MATERIALS:
7.1 SOURCE AND COLLECTION OF DATA :

Data will be generated using laboratory experimental techniques on animals evaluations such as antisecretory and antiulcer activities on albino rats of either sex. The results data obtained from the present study shall be analyzed by one-way analysis of variance (ANOVA) test followed by Dunnett,s t test.




    1. PHARMACOLOGICAL STUDIES:


Animals:

Albino rats weighing between 150-200g will be used and randomly assigned into 3 groups of 6 animals each one of control, second of standard and third of test.


Evaluation for acute toxicity study:23

The albino mice of either sex will be used during investigation. The animals are fasted over night prior to the experimental procedure. The OECD guidline no-420 fixed dose method will be adopted and accordingly doses of extract will be caluculated. 1/10 of the LD 50 will be administered.




  1. Pylorus ligated model 24 :

In this method, albino rats will be deprived of food for 24 hr, care is taken to avoid coprophagy. Under light ether anesthesia the abdomen is opened by small midline incision and pyloric end portion of the stomach is ligated .The animals would be killed, 4 hr after the operation under the over dose of anesthetic ether and the gastric contents would be collected. After centrifugation, the gastric content of each animal will be individually assessed for volume of gastric secretion. Free and total acid production will be estimated by titrating against 0.01N NaOH using Topfer’s reagent and phenolphthalein as an indicator. Drugs/extract will be administered 30 min prior to pylorus ligation. Effective dose of the one is administered to the rest of other groups.


b) Cold stress induced ulcer model 25:

In this method, rats will be divided into 3 groups of 6 animals each one of control, second of standard and third of test. In this method, wistar rats will be deprived of food for 12 hrs, they are then immobilized in stress cage and forced to remain in a cold room (4-6˚C) for 3 hrs. The animals would be sacrificed and ulcer index is calculated. Drugs/extract will be administered 30 min prior to cold stress.


c) Aspirin induced ulcer model 26:

The rats will be divided into 3 groups of 6 animals each one of control, second of standard, and third of test. Aspirin will be suspended in 1% carboxy methylcellulose in water (20mg/ml) and administered orally in a dose of 500mg/kg in 36 hrs, fasted rats. Four hours later the animals are sacrificed and ulcer index is calculated. Drugs/extract will be administered 30 min prior to aspirin administration.


c) Ethanol induced ulcer model 27:

The albino rats will be divided into 3 groups of 6 animals each one of control, second of standard, and third of test. Ethanol will be orally administered at a dose of 5ml/kg. The animals were killed 15 minutes after ethanol administration. The gastric lesions would be evaluated. Drugs/extract will be administered 30 min prior to ethanol administration.



7.3 Does the study require any investigation or interventions to be conducted on patients or other humans/animals? If so please described briefly.

The above study requires investigation to be done on albino rats of wistar strain for the determination of gastric antisecretary and antiulcer activities. These studies are planned in accordance with the procedure reported in the literature.




    1. Has ethical clearance been obtained from your institution in case?

The study has been given for clearance from the ethical committee of the institution.



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REFERENCES:


  1. Harshmohan. Textbook of Pathology, 4th ed. 2004. Jaypee brother Medical publisher Ltd, New Delhi.

  2. K.D.Tripathi. Essentials of Medical Pharmacology” 5th ed. Jaypee brother Medical publisher Ltd, New Delhi.

  3. Anoop A, Jagadeesan M. Biochemical studies on the anti-ulcerogenic potential of hemidesmus indicus R.Br.Var. Indicus. J. Ethanopharmacol 2003;84:149-6.

  4. Khare CP. 2004. Encyclopedia of Indian medicinal plants. Springer-verlag berlin heidol: 309-0.

  5. Arya Vaidya Sala. Indian medicinal plants. 4th ed.1995. orient longaman private Ltd.

  6. Kirtikar K R, Basu B D, Indian medicinal plants, 2nd ed. Allahabad, India;1998.

  7. The wealth of India, raw materials, 4th ed.

  8. Ka MH, choi EH, Chun HS, Lee KG. Antioxidant activity of volatile extracts isolated from Angelica tenuissimae roots, peppermint leaves, pine needles, and sweet flag leaves. J Agric Food Chem 2005;53(10):4124-9.

  9. Dorman HJ, Kosar M, Kahlos K, Holm Y, Hiltunen R. Antioxidant properties and composition of aqueous extracts from Mentha species, hybrids, varieties, and cultivars. J Agric Food Chem 2003;51(16):4563-9.

  10. Baliga MS, Jagetia GC, Rao SK, Babu K. Evaluation of nitric oxide scavenging activity of certain spices in vitro: a preliminary study. 2003; 47 (4): 261-4.

  11. Shin TY. Inhibition of immunologic and nonimmunologic stimulation-mediated anaphylactic reactions by the aqueous extract of Mentha arvensis Linn. Immunopharmacol Immunotoxicol 2003;25(2):273-3.

  12. Koreamed. Inhibition of IgE-mediated anaphylactic reaction by Mentha arvensis Linn. in rats, J asthma allergy Clin Immunol 2000;20(4):601-8.

  13. Marta Cristina Teixeira Duarte, Glyn Mara Figueira, Adilson Sartoratto, Vera Lucia Garcia Rehder and Camila Delarmelina. Anti-candida activity of brazilian medicinal plants. J Ethanopharmacol 2005;97(2):305-11.



  1. Nidhi Sharma and D. Jocob. Antifertility investigation and toxicological screening of the petroleum ether extract of the leaves of Mentha arvensis Linn. in male albino mice. J Ethanopharmacol 2001;75(1):5-12.

  2. Savita V. Phatak and Mohan R. Heble. Organogenesis and terpenoid synthesis in Mentha arvensis Linn. Fitoterapia 2002;73(1):32-9.

  3. Bhusita Wannissorn, Siripen Jarikasem. Thammathad Siriwangchai and Sirinum Thubthimthed. Antibacterial properties of essential oils from Thai medicinal plants. Fitoterapia, 2005;76(2):233-36.

  4. Attia H. Atta and Samar M. Mouneir. Antidiarrhoeal activity of some Egyptian medicinal plant extracts.J ethanopharmacol 2004;92(2-3):303-09.

  5. Usha Kiran and D. D Patra. Medicinal and aromatic plant materials as nitrification inhibitors for augmenting yield and nitrogen uptake of Japanese mint (Mentha arvensis Linn.).Bioresourse Technol 2003;86(3):267-76.

  6. Imai H, Osawa K, Yasuda H, Hamashima H, Arai T, Sasatsu M. Inhibition by the essential oils of peppermint and spearmint of the growth of pathogenic bacteria. Microbios 2001;106(1):31-9.

  7. Primorac M, Sekuovic D, Antonic S. In vitro determination of the spermicidal activity of plant saponins. Pharmazie1985;40(8):585.

  8. Sharma N, Jacob D. Assessment of reversible contraceptive efficacy of methanol extract of Mentha arvensis Linn. leaves in male albino mice. J Ethanopharmacol 2002;80(1):9-13.

  9. Mathur R. Fructolysis effect of 50% ethanolic extract of Mentha arvensis Linn. (Leaves) in seminal vesicles of rat. Acta Eur Fertil 1991;22(4):219-20.

  10. OECD( Organisation for Economic Co-operation and Development). Guideline 420: Acute oral toxicity-Fixed dose Procedure, Paris: OECD;1992.

  11. Kanjanapothi D, Smitasiri Y, Panthong A, Taesotikul T, Rattanapanone V. Postcoital antifertility effect of Mentha arvensis Linn. Contraception 1981;24 (5):559-67.


  1. Shay M, komarav SA, Fels, Meranza D, Gruenstein H, Siplet H . A simple method for uniform production of gastric ulceration in rats, Gastroenterol 1945; 5:43-1.

  2. Vincent GP, Glavin GB, Rutkowski JL, pare WP. Body orientation, food deprevation and potention of restraint ulcers induced gastric lesions, Gastroent. Clin Biol, 1977;1:539-3.

  3. Ganguly AK, Bhatnagar OP. Effect of bilateral adrenalectomy on the production of restraint ulcers in stomach of albino rats. Canad j physiol & pharmacol 1973;51:748-0.

  4. Chi- Feng Liu, Chun-Chung Lin, Lean Tienk Ng, Song-Chow Lin. Protection by Teramethylpyrazine in acute absolute ethanol induced gastric lesions, J Biomed sci 2002;9:395-0.



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