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Research Gate: Pharmaceutical Sciences, 1 (2013) 76-79 Antiulcerogenic Effects of Terminalia chebula (Retz.) in Pylorus Ligated Rats
1Department of Pharmaceutical Sciences, Doctor Hari Singh Gour Vishwavidyalaya Sagar-(M.P.) India 2MES College of Pharmacy Sonai (M.H.) India A R T I C L E I N F O
A B S T R A C T
Keywords:
The cause of ulceration in patients is mainly due to hyper secretion of gastric juice and also due to hyper secretion of pepsin. In traditional system of medicine a number of herbal preparations have been used for the treatment of peptic ulcers. The marketed poly-herbal formulation and many synthetic drugs has been used for the treatment of gastric-ulcer diseases. In view of this, in present study we have to evaluate the anti-ulcer activity of Terminalia chebula. Study wascarried out, on pyloric ligation induced ulcers in rats pretreated with the doses of 1ml/kgAbsolute ethanol, Extracts and fraction of Terminalia chebula, Omeprazole: 20 mg/kg body wt.
After 4 hour in pyloric ligation method induced method rat was sacrificed and stomach wasremoved for observation of various biochemical and histo-pathological parameters such as pH ofgastric content, free acidity, total acidity and ulcer index were determined. The results ofpharmacological activity led to the conclusion that the Alcoholic extract and Ethyl acetatefraction of alcoholic extract exhibited activity that is more significant. Ethanol extract and its Article History:
Ethyl acetate fraction significantly raised the pH of gastric content and thus reducing the level of acidity in the stomach. It lowered the free and total acidity and ulcer index as compared to control group. The antigastric-ulcer activity may be due to Gallic acid as the fraction contains Introduction
effects and limitations Herbal medicine deals with plants The acid-peptic diseases are those disorders in which and plant extracts in treating diseases. These are gastric acid and pepsin are necessary, but usually not considered safer because of the natural ingredients with sufficient, pathogenic factors. While inherently caustic, no side effects. Gastric ulcer, one of the most widespread, acid and pepsin in the stomach normally do not produce is believed to be due to an imbalance between aggressive damage or symptoms because of intrinsic defense and protective factors.2 The gastric mucosa is mechanisms. Barriers to the reflux of gastric contents into continuously exposed to potentially injurious agents such the esophagus comprise the primary esophageal defense.
as acid, pepsin, bile acids, food ingredients, bacterial If these protective barriers fail and reflux occurs, products (Helico bacter pylori) and drugs.3 dyspepsia and/or erosive esophagitis may result. 1 Ulcers These agents have been implicated in the pathogenesis of are sores on the lining of the digestive tract which consists gastric ulcer, including enhanced gastric acid and pepsin of the esophagus, stomach, duodenum and intestines. Most secretion, inhibition of prostaglandin synthesis and cell ulcers are located in the duodenum and they are called proliferation growth, diminished gastric blood flow and duodenal ulcers. Ulcers located in the stomach are called gastric ulcers. Ulcers in the esophagus are called Terminalia chebula (Family: Combretaceae) was one of the traditional medicine used in many folk claims and it is Stomach ulcer (peptic ulcer) is a breaking in the tissue called as “King of medicine”. It is an middle-sized tree lining of the stomach. Most are caused by infection with leaves are ovate, or elliptic, flowers are yellowish white, the Helicobacter pylori bacterium and not by stress or fruits are yellowish brown in colour distributed poor diet. Symptoms may include abdominal pain just throughout in India.5 The plant has extensively used in below the ribcage, indigestion, nausea, loss of appetite, ayurveda and sidda for Constipation, diarrhea, ulcers, vomiting, weight loss, blood in the vomit or bowel motions gastroenteritis, asthma, cough, dyspnea, dyspepsia, and symptoms of anaemia, such as light-headedness. The hemorrhoids, candidiasis, parasites, malabsorption existing treatment options include antibiotics and acid- syndrome, hepatomegaly, vesicular and renal calculi, urinary discharges, tumors, skin diseases, leprosy, Although a number of antiulcer drugs such as H antagonists, proton pump inhibitors and cytoprotectants neuropathy, paralysis, memory loss, epilepsy, depression, are available for ulceration all these drugs have side diabetes, cardiovascular diseases, anorexia, wounds.
* Corresponding author: [email protected] Research Gate: Pharmaceutical Sciences, 1 (2013) A peptic ulcer, also known as ulcus pepticum or peptic Pylorus Ligation induced gastric ulcers
ulcer disease, is an ulcer (defined as mucosal erosions equal to or greater than 0.5 cm) of an area of the Albino rats were divided into five groups of six animals gastrointestinal tract that is usual y acidic and thus each and fasted for 48h with free access to water. Pyloric extremely painful. 70-90% of ulcers are associated with ligation was performed under light ether anesthesia to Helicobacter pylori, a spiral-shaped bacterium that lives in each animal. Animals were given 1% Tween 80 solution or the acidic environment of the stomach. Ulcers can also be extracts 250mg/ kg or 20 mg/ kg omeprazole orally caused or worsened by drugs such as aspirin and other immediately after pylorus ligation. Animals were sacrificed 4 h later. The stomach was carefully removed and gastric contents were collected. The excised stomach Therapies are directed at decreasing gastric acidity, was studied histologically and the gastric juice was enhancing the lower esophageal sphincter, or stimulating centrifuged at 3000 rpm for 30 min and the supernatant esophageal motility. In the stomach, mucus and was used for analysis of various biochemical and bicarbonate, stimulated by the local generation of prostaglandins, protect the gastric mucosa. If these defenses are disrupted, a gastric or duodenal ulcer may Biochemical Parameters:
form. The treatment and prevention of these acid-related Estimation of pH of gastric contents
disorders are accomplished either by decreasing the level of gastric acidity or by enhancing mucosal protection.
The pH of Gastric contents were measured by pH meter, Materials and Methods
Table 1: pH Study of Rat Gastric juice treated with
Plant Material
extracts/fraction of fruit of Terminalia chebula Fruits of Terminalia chebula were purchased from local Alcoholic
Ethyl acetate
market of Sagar (M.P.) in the months of November and Fraction
December 2011. Fruits were authenticated by Dr. P.
Tiwari (Herbarium no. Bot/1019) at the Department of
Mean ±SD (n=6 Significance * p<0.01); #std. Omeprazole
Botany Dr. H. S. Gour Vishwavidyalaya, Sagar (M.P.) and preserved in the herbarium of the institute.
Determination of free acidity and total acidity of
Gastric contents
Preparation of Extracts
The gastric contents were centrifuged at 1000 rpm for Successive Solvent Extraction
10min. 1ml of supernatant was diluted with 9ml of The powder of fruits (600g) were successively extracted distilled water. A volume of 2ml diluted gastric juice was with petroleum ether (60-80C), benzene, ethyl acetate, titrated with 0.1N Sodium hydroxide run from a chloroform, ethanol and water by soxhlet extraction microburette using 3-4 drops of Topfer's reagent as process. During the extraction, the test was performed for indicator until canary yellow colour was observed. Volume the completion of the process. The completion of the of NaOH required was noted. This corresponds to free extraction was confirmed by evaporating a few drops of acidity. Further 2-3 drops of phenolphthalein was added the extract on the watch glass and ensuring that no and titrated with NaOH until pink colour was restored.
residue is found after evaporating the solvent.7 The marc This gives total acidity. Free acidity and total acidity is was dried in hot air oven below 40C before being expressed in terms of ml of 0.1N HCl per 100 gms of extracted with the next solvent. The solvent was gastric contents. This is the same as mEq/lit. To obtain recovered by distillation, and the last traces of solvent this figure multiply the burette reading obtained from were removed under vacuum. The extract obtained with titration by 10 (Table 2 and 3). The total acidity is each solvent was weighed to a constant weight and expressed as m Eq/L by the following formula: percentage on w/w basis was calculated.
Acidity = (Vol. of NaOH × N × 100)/0.1 mEq/L Drugs and Chemicals
Table 2: Free acidity of Rat Gastric contents of treated
Omeprazole drug sample was received from Sun Pharma with fruit extracts/fraction of Terminalia chebula and used during the experimental protocol.
Alcoholic
Alcoholic
Fraction
In vivo Study
Animal selection-Wistar albino rats
Mean ±SD (n=6 Significance * p<0.01); #std. Omeprazole
Wistar albino rats weighing between 150 and 200 g were Table 3: Estimation of Total acidity of Gastric contents
selected for the antiulcer studies. The animals were acclimatized to standard laboratory conditions of Alcoholic
Alcoholic
Fraction
temperature (22±30°C) and maintained on 12:12 h light:dark cycle and distilled water. The animal care and experimental protocol was in accordance with IAEC of Department of Pharmaceutical sciences, Dr. H. S. Gour Mean ±SD (n=6 Significance * p<0.01); #std. Omeprazole
Research Gate: Pharmaceutical Sciences, 1 (2013) Histological Parameters
a known anti-ulcer drug, Ethyl acetate fraction of Alcoholic Ulcer Index study and Percentage Inhibition
extract and alcoholic extract showed good results, whereas Aqueous extract are less potent in decreasing Rat stomach were excised and opened along the greater gastric acidity. The results are tabulated in Table 2.
curvature, washed and stretched and studied for its Estimation of total acidity of gastric content in terms
histology. The internal surface of stomach was examined of ml of 0.1N HCl / 100ml of gastric content
for the presence of lesions and the extent of the lesions was measured. The number of the lesions along the wall of Gastric total acidity is increased to (44.67 mEq/litre) in stomach was recorded as ulcer index and % inhibition was control animals. Ethyl acetate fraction (34.87 mEq/litre) also calculated (Basile, et al., 1990). Each stomach sample showed significant decrease in total acidity (p<0.01) as was examined grossly and the ulcers were graded using compared to control. The decrease in total acidity by the following system suggested by Kunchandy et al. 1985.
alcoholic extract and Aqueous extract is 41.4 and 42.56 mEq/litre) respectively. The results are tabulated in Table Calculation of Ulcer index (UI) and percent inhibition
(% I)
Determination of ulcer index
Ulcer Index (UI) = (Number of ulcers in control- Number The ulcer index in control animals was (5.18). Ethyl acetate Fraction (3.85) significantly reduced the ulcer Percent Inhibition (% I) =(UI of control – UI of test *100)/ index (p<0.01) as compared to control. The reduction in ulcer index by alcoholic extract and aqueous extract is 4.4 and 4.85 respectively. Omeprazole, a standard anti-ulcer Statistics -
drug showed ulcer index 2.19. The results are tabulated in The ulcer index and percent ulcer inhibition and Statistical analysis was performed by one-way analysis of variance Table 4: Ulcer Index and ulcer Inhibition percentage
(ANOVA) followed by Dunnett’s t-test for multiple profile of fruit extracts/fraction of Terminalia chebula comparisons. The significance of difference was accepted Parameters
Alcoholic
Alcoholic
at p < 0.05. An analysis was performed using the software Fraction
Observation
The prepared slides were viewed under light microscope #std. Omeprazole
(100X). The observed stomach epithelium was Red colored and severely eroded with perforations in case of Conclusion
control group of animals. The number of ulcers was more The pathogenesis of gastric ulcers remains still unclear, in control group as compared to the treated groups. The ranges from stressful lifestyle, alcohol consumption, use of number of ulcers and severity of ulceration was less in steroidal and non-steroidal anti inflammatory drugs groups treated with extracts and Ethyl acetate fraction.
(NSAIDs) and drugs which stimulate gastric acid and The presence of few ulcer sites and ulceration of moderate pepsin secretion, Helicobacter pylori infections, smoking, or negligible nature in case of Ethyl acetate fraction lower socio-economic status and family history. All these treated groups of animals gives significant indication of represent significant risk factors that may contribute to potent ulcer protective effects compared to control increasing gastric damage. Now days, various approaches have been made to study herbal drugs for treatment of various gastrointestinal disorders. From the present study, it was concluded that all extracts possesses Estimation of pH of gastric content
significant anti-ulcerogenic effect at varied level, thus a In control animals, without any drug the mean is 2.51.
specific chemical entity which is responsible for the anti- Ethyl acetate Fraction showed significant rise in pH (3.37) ulcerogenic effect is yet to be explored and suggested by as compared to control. The rise in pH shown by Alcoholic the authors at molecular level by altering protocols at and aqueous extracts are 2.71 and 2.62 respectively.
Omeprazole, a standard drug raised the pH to 3.86, which Reference
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