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Anti-ulcer and anti-oxidant activity of Pepticare, a herbomineral formulation.

Abstract

Pepticare, a herbomineral formulation of the Ayurveda medicine consisting of the herbal drugs: Glycyrrhiza glabra, Emblica officinalis and Tinospora cordifolia, was tested for its anti-ulcer and anti-oxidant activity in rats. Effects of various doses (125, 250, 500 and 1000 mg/kg, p.o.) of Pepticare were studied on gastric secretion and gastric ulcers in pylorus-ligation and on ethanol-induced gastric mucosal injury in rats. The reduction in ulcer index in both the models along with the reduction in volume and total acidity, and an increase in the pH of gastric fluid in pylorus-ligated rats proved the anti-ulcer activity of Pepticare. It was also found that Pepticare was more potent than G. glabra alone in protecting against pylorus-ligation and ethanol-induced ulcers. The increase in the levels of superoxide dismutase, catalase, reduced glutathione and membrane bound enzymes like [Ca.sup.2+] ATPase, [Mg.sup.2+] ATPase and [Na.sup.+] [K.sup.+] ATPase and decrease in lipid peroxidation in both the models proved the anti-oxidant activity of the formulation. Thus it can be concluded that Pepticare possesses anti-ulcer activity, which can be attributed to its anti-oxidant mechanism of action.

[c] 2004 Elsevier GmbH. All rights reserved.

Keywords: Anti-ulcer; Anti-oxidant; Lipid peroxidation; Superoxide dismutase; Catalase; Reduced glutathione; Ayurveda medicine; Glycyrrhiza glabra; Emblica officinalis; Tinospora cordifolia

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Introduction

Peptic ulcer is the most common gastrointestinal disorder in clinical practice. Considering the several side effects (arrythmias, impotence, gynaecomastia and haematopoeitic changes) of modern medicine, indigenous drugs possessing fewer side effects should be looked for as a better alternative for the treatment of peptic ulcer (Akhtar et al., 1992).

There is evidence concerning the participation of reactive oxygen species in the etiology and pathophysiology of human diseases, such as neurodegenerative disorders, inflammation, viral infections, autoimmune pathologies and digestive system disorders such as gastrointestinal inflammation and gastric ulcer (Repetto and Llesuy, 2002). Studies have shown alterations in the anti-oxidant status following ulceration, indicating that free radicals seem to be associated with the pylorus ligation-induced (Rastogi et al., 1998) and ethanol-induced (Pihan et al., 1987; Mizui et al., 1987) ulceration in rats. Drugs with multiple mechanisms of protective action, including anti-oxidant properties, may be one way forward in minimizing tissue injury in human disease (Barry, 1991).

Since decades many indigenous drugs have been known to posses anti-ulcer activity. The anti-ulcer properties of Emblica officinalis (Rajeshkumar et al., 2001; Al-Rehaily et al., 2002) and Glycyrrhiza glabra (De et al., 1997) have been mentioned. The anti-oxidant properties of Tinospora cordifolia (Mathew and Kuttan, 1997; Prince and Menon, 2001.), E. officinalis (Scartezzini and Speroni, 2000; Bhattacharya et al., 1999; Mathur et al., 1996), G. glabra (Hatano et al., 1991) and Suvarna Makshik bhasma (Shah and Vohora, 2002; Mitra et al., 2002) were earlier investigated and were found to possess free radical scavenging property. These ingredients were also found to produce significant induction in the levels of various endogenous antioxidant enzymes.

It has been demonstrated that many drugs or formulations possess potent anti-oxidant actions and are effective in healing experimentally induced gastric ulcers. From these findings, Pepticare is expected to be effective in preventing ulcer formation and in ulcer healing. In addition in order to clarify whether or not Pepticare exerts an anti-ulcer action by means of its antioxidant activity, we examined the effect of this formulation on the content of thiobarbituric acid-reactive substances, an index of lipid peroxidation and on the activities of endogenous anti-oxidant enzymes.

The present study was thus aimed to investigate the anti-ulcer effects of Pepticare along with its effect on the anti-oxidant enzymes to justify whether the formulation exerts an anti-ulcer action by means of its anti-oxidant activity.

Material and methods

Composition

Each gram of Pepticare (manufactured by Ayur Herbals Pvt. Ltd., Baroda, India) contains powders of T. cordifolia Miers (Menispermaceae; Whole plant; 300 mg), E. officinalis Gaertn. (Euphorbiaceae; Fruit; 200 mg), G. glabra Linn. (Papilionaceae; Root; 300 mg) alongwith Sootshekhar ras (40 mg), Praval bhasma (Corallium rubram; 30 mg), Suvarna Makshik bhasma (Ferri sulphuratum; 20 mg), Kapardi bhasma (calcium; 80 mg) and Shodhit gairik (silicate of alumina and oxide of iron; 30 mg).

Animals

Female albino rats of wistar strain weighing between 150 and 225 g were used for the study. The animals were fed ad libitum with standard pellet diet and had free access to water.

Experimental procedure

The animals were divided into nine groups, each consisting of six rats. Group I represented the control group, which received 5 ml/kg body weight of vehicle (1% gum acacia, p.o.). Groups 2-5 received Pepticare orally at the doses of 125, 250, 500 and 1000 mg/kg body weight, respectively. Groups 6 received powder of G. glabra orally at the dose of 150 mg/kg body weight.

Study of anti-ulcer activity using pylorus ligation method

The method of Shay rat ulcer (Shay et al., 1945) was adopted. Rats were fasted for 48 h. The drug, Pepticare or G. glabra was administered to the animals. During the course of the experiment, food was withdrawn. After the pretreatment period of 1 h, the animals were anaesthetised with anaesthetic ether. The abdomen was opened by a small midline incision below the xiphoid process; pylorus portion of stomach was slightly lifted out and ligated. Precaution was taken to avoid traction to the pylorus or damage to its blood supply. The stomach was placed carefully in the abdomen and the wound was sutured by interrupted sutures. Nineteen hours after pylorus ligation the rats were sacrificed and the stomach was removed. The gastric content was collected and centrifuged. The volume, pH and total acidity of gastric fluid was determined. The stomach was then incised along the greater curvature and observed for ulcers. The number of ulcers was counted using a magnifying glass and the diameter of the ulcers was measured using a vernier caliper. Ulcer index was determined by following the scoring method of Suzuki et al. (1976).

Score 1: maximal diameter of 1 mm.

Score 2: maximal diameter of 1-2 mm.

Score 3: maximal diameter of 2-3 mm.

Score 4: maximal diameter of 3-4 mm.

Score 5: maximal diameter of 4-5 mm.

Score 10: an ulcer over 5 mm in diameter.

Score 25: a perforated ulcer.

Study of anti-ulcer activity using ethanol-induced ulcer method

Pepticare or G. glabra was administered orally to the rats for a period of 10 days. On the 10th day, 1 h after the final dose of Pepticare or G. glabra, 96% ethanol (5 ml/kg, p.o.) was administered to the overnight fasted rats of all groups. The animals were then sacrificed 1 h after the dose of ulcerogen. The stomach was then removed, incised along the greater curvature and its mucosal erosion was determined randomly by measuring the area of the lesions. The sum of the areas was expressed as ulcer index ([mm.sup.2]).

Study of anti-oxidant activity of Pepticare

The stomach of rats of Group 1 (Control) and Groups 2-5 (Pepticare-treated groups) was then weighed and homogenized in chilled Tris buffer (10 mM, pH 7.4) at a concentration of 10% w/v. The homogenates were centrifuged at 10,000g at 0[degrees]C for 20 min using Remi C-24 high-speed cooling centrifuge. The clear supernatant was used for the assays of lipid peroxidation (MDA content), endogenous anti-oxidant enzymes (superoxide dismutase and catalase) and reduced glutathione (GSH). The sediment was resuspended in ice cold Tris buffer (10 mM, pH 7.4) to get a final concentration of 10% and was used for the estimation of different membrane bound enzymes ([Na.sup.+][K.sup.+] ATPase, [Ca.sup.2+] ATPase and [Mg.sup.2+] ATPase) and proteins.

Superoxide dismutase (SOD) was determined by the method of Mishra and Fridovich (1972). Catalase was estimated by the method of Hugo Aebi as given by Colowick et al. (1984). Reduced glutathione was determined by the method of Moron et al. (1979). Lipid peroxidation or malondialdehyde (MDA) formation was estimated by the method of Slater and Sawyer (1971). Membrane bound enzymes namely [Na.sup.+][K.sup.+] ATPase, [Ca.sup.2+] ATPase and [Mg.sup.2+] ATPase were assayed according to the methods of Bonting (1970), Hjerten and Pan (1983) and Ohnishi et al. (1982), respectively. The inorganic phosphorus was estimated by the method of Fiske and Subbarow (1925). Total proteins were determined by the method of Lowry et al. (1951).

Statistical analysis

Results of all the above estimations have been indicated in terms of mean[+ or -]SEM. Difference between the groups (Pepticare and control groups) was statistically determined by analysis of variance followed by Tukey-Kramer Multiple Comparisons test, with the level of significance set at p<0.05. G. glabra group was compared with the control group by using unpaired Student's t-test.

Results

Study of anti-ulcer activity using pylorus ligation method

It was observed that in the vehicle treated control group the ulcer index was 92.75[+ or -]5.20 and the maximum number of ulcers were of the ulcer score 4 and 5. In the rats of this group a number of perforated ulcers (score 25) were also observed.

Pepticare was found to produce a decrease in ulcer index at all the four doses; the percentage reduction being 16.77%, 48.46%, 70.46% and 92.24%, respectively. Significant reduction (p<0.001) in ulcer index was observed at the doses of 250, 500 and 1000 mg/kg. All the ulcers were of score 1, 2, 3 and 4 and no perforated ulcers were observed. The formulation also significantly reduced the volume and total acidity, and increased the pH of the gastric fluid, proving its anti-ulcer activity (Table 1).

G. glabra (150 mg/kg) was not found to produce a significant reduction in ulcer index. It neither reduced the total acidity and volume significantly nor increased the pH of gastric fluid at the dose of 150 mg/kg (Table 4).

Study of anti-ulcer activity using ethanol-induced ulcer method

Administration of ethanol produced significant ulcers (287.98[+ or -]17.79) in the control group. There was a significant (p<0.001) reduction in ulcer index at all the four doses of Pepticare by 72.57%, 85.93%, 94.31% and 98.44%, respectively (Table 3); whereas G. glabra at the dose of 150 mg/kg did not produce a significant reduction in ulcer index (Table 4).

Study of anti-oxidant activity of Pepticare

Pylorus-ligation was found to increase lipid peroxidation and decrease SOD, catalase and reduced glutathione in the control group, thus leading to oxidative stress. Administration of Pepticare, at the doses of 250, 500 and 1000 mg/kg, brought about a significant reduction in lipid peroxidation and an increase in the content of reduced glutathione. The activities of antioxidant enzymes namely, SOD and catalase were also found to be significantly increased at the dose of 1000 mg/kg. An enhancement in the membrane bound ATPases was also observed by Pepticare (Table 2).

Ethanol administration was found to increase lipid peroxidation and decrease SOD, catalase and reduced glutathione in the control group. Administration of Pepticare at the doses of 250, 500 and 1000 mg/kg significantly decreased lipid peroxidation and increased the glutathione content and the activities of SOD and catalase. All the membrane bound ATPases were also found to be elevated in the drug treated groups (Table 3).

Discussion and conclusion

The present study demonstrates that Pepticare exhibits both gastroprotective and ulcer healing properties, probably as a result of the anti-oxidant action of the drug.

Although in most of the cases the aetiology of ulcer is unknown, it is generally accepted that it results from an imbalance between aggressive factors and the maintenance of the mucosal integrity through the endogenous defence mechanism (Piper and Stiel, 1986). To regain the balance, different therapeutic agents including herbal preparations are used to inhibit the gastric acid secretion or to boost the mucosal defence mechanism by increasing mucus production.

Pepticare, a herbal drug formulation, consists of plants that are mentioned in Indian system of medicine (Ayurveda) for their remedial properties. The anti-ulcer effect of Pepticare was tested against gastric lesions induced by pylorus-ligation and ethanol, the experimental models related to lesion pathogenesis with production of reactive species. Pepticare prevented the mucosal lesions induced by pylorus-ligation and ethanol. Pepticare was also found to increase the pH and decrease the acid volume and total acidity of gastric fluid. These effects of Pepticare treatment on the parameters that influence the initiation and induction of ulceration may be considered as highly desirable property of anti-ulcerogenic agent.

Considering the amount of Glycyrrhiza glabra powder for which an anti-inflammatoric (corticomimetic) effect has been proven and which is present in the formulation, i.e. after comparing the ulcer index of 500 mg/kg of Pepticare with that of 150 mg/kg of G. glabra in both the models, it was found that Pepticare was 8.5 and 51 times more potent than G. glabra in protecting against pylorus-ligation and ethanol-induced ulcers, respectively. Thus the study proves the therapeutic advantage of Pepticare over G. glabra alone. It can thus be said that the herbal mixture with the minerals has a much better anti-ulcer effect than Glyzyrrhiza alone, due to potentiated synergistic effects.

Reactive oxygen species are involved in the pathogenesis of pylorus ligation-induced (Rastogi et al., 1998) and ethanol-induced (Pihan et al., 1987) gastric mucosal injury in vivo. Results in the present study also indicate similar alterations in the anti-oxidant status after pylorus ligation and ethanol induced ulcers.

Emblica officinalis and Tinospora cordifolia are categorized as 'rasayanas' (rejuvenatives). Rasayanas are non-toxic Ayurvedic complex herbal preparations or individual herbs used to rejuvenate or attain the complete potential of an individual in order to prevent diseases and degenerative changes that leads to disease. Various activities of rasayanas have been reviewed by Vayalil et al. (2002) to support the above concept, its role as a prophylactic medication and significance in the prevention of diseases in both healthy as well as diseased individuals. The emerging data suggest that the possible mechanisms may be by immunostimulation, quenching free radicals, enhancing cellular detoxification mechanisms, repair damaged non-proliferating cells, inducing cell proliferation and self-renewal of damaged proliferating tissues, and replenishing them by eliminating damaged or mutated cells with fresh cells. The clinical efficacy of the fruits of E. officinalis is held in high esteem in Ayurveda and amla is referred to as a 'maharasayana'. By virtue of their properties and clinical use in Ayurveda, the rasayanas may provide potential therapeutic intervention against oxidative threats, both in health and disease.

Preventive anti-oxidants, such as superoxide dismutase (SOD) and catalase (CAT) enzymes are the first line of defence against reactive oxygen species. Reduced glutathione (GSH) is a major low molecular weight scavenger of free radicals in the cytoplasm and an important inhibitor of free radical mediated lipid peroxidation (Halliwell, 1995). Administration of Pepticare resulted in a significant increase in the SOD, catalase and reduced glutathione levels as compared to the control animals, which suggests its efficacy in preventing free radical-induced damage.

Lipid peroxidation is a free radical mediated process, which has been implicated in a variety of disease states. It involves the formation and propagation of lipid radicals, the uptake of oxygen and rearrangement of double bonds in unsaturated lipids which eventually results in destruction of membrane lipids. Biological membranes are often rich in unsaturated fatty acids and bathed in oxygen-rich metal containing fluid. Therefore it is not surprising that membrane lipids are susceptible to peroxidative attack (Cheesman, 1993). The study has revealed a significant decrease in lipid peroxidation by Pepticare in both the experimental models, which suggests its protective effect.

[Na.sup.+][K.sup.+] ATPase, [Ca.sup.2+] ATPase and [Mg.sup.2+] ATPase are membrane bound enzymes. The drug significantly increased the activity of all the ATPases in both the models.

It is thus concluded that Pepticare is an effective anti-ulcer agent. Further, this study also proves that the anti-ulcer effect may be due to its anti-oxidant mechanism of action.
Table 1. Effect of Pepticare on the various gastric parameters of
pylorus-ligated rats

Groups Ulcer index Volume of gastric fluid
 (ml)

Control 92.75[+ or -]5.20 17.28[+ or -]1.18
Pepticare (125 mg/kg) 77.21[+ or -]3.93 (NS) 17.25[+ or -]0.87 (NS)
 (16.77%)
Pepticare (250 mg/kg) 47.83[+ or -]7.40*** 14.88[+ or -]1.82 (NS)
 (48.46%)
Pepticare (500 mg/kg) 27.44[+ or -]3.53*** 11.50[+ or -]0.68*
 (70.46%)
Pepticare (1000 mg/kg) 7.22[+ or -]1.50*** 10.50[+ or -]0.32**
 (92.24%)
F value 53.332 7.739

Groups pH of gastric fluid Total acidity (mEq/1/
 100 g)

Control 1.20[+ or -]0.07 127.01[+ or -]3.63
Pepticare (125 mg/kg) 1.58[+ or -]0.07 (NS) 112.5[+ or -]3.52*
Pepticare (250 mg/kg) 1.85[+ or -]0.56* 89.03[+ or -]2.68***
Pepticare (500 mg/kg) 2.05[+ or -]0.19** 61.06[+ or -]1.68***
Pepticare (1000 mg/kg) 3.25[+ or -]0.11*** 31.74[+ or -]3.21***
F value 36.423 161.93

Values are expressed as mean [+ or -] SEM.
Pepticare treated groups were compared with control group.
*p < 0.05; **p < 0.01; ***p < 0.001; NS = non significant.
Values in parenthesis indicate the % reduction in ulcer index in
relation to the control group.

Table 2. Effect of Pepticare on the antioxidant parameters in stomach of
pylorus ligated rats

Parameters Normal Control

SOD (Unit/mg protein) 5.64[+ or -]0.33 2.37[+ or -]0.26***
Catalase ([micro]moles 8.27[+ or -]0.27 5.93[+ or -]0.61*
 of [H.sub.2][O.sub.2]
 consumed/min/mg protein)
Reduced glutathione 3.31[+ or -]0.15 0.58[+ or -]0.14***
 ([micro]g of GSH/mg
 protein)
Lipid peroxidation (nmoles 3.45[+ or -]0.24 10.10[+ or -]0.43***
 of MDA/mg protein)
[Na.sup.+] [K.sup.+] 5.29[+ or -]0.23 1.49[+ or -]0.11***
 ATPase ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Ca.sup.2+] ATPase 3.65[+ or -]0.12 1.63[+ or -]0.62***
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Mg.sup.2+] ATPase 3.52[+ or -]0.36 1.53[+ or -]0.63***
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)

Parameters Pepticare
 125 mg/kg 250 mg/kg

SOD (Unit/mg protein) 2.42[+ or -]0.45 (NS) 2.78[+ or -]0.37 (NS)
Catalase ([micro]moles 5.96[+ or -]0.64 (NS) 6.49[+ or -]0.43 (NS)
 of [H.sub.2][O.sub.2]
 consumed/min/mg protein)
Reduced glutathione 1.51[+ or -]0.24 (NS) 2.71[+ or -]0.58***
 ([micro]g of GSH/mg
 protein)
Lipid peroxidation (nmoles 9.12[+ or -]0.60 (NS) 5.87[+ or -]0.35***
 of MDA/mg protein)
[Na.sup.+] [K.sup.+] 1.50[+ or -]0.05 (NS) 1.92[+ or -]0.17 (NS)
 ATPase ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Ca.sup.2+] ATPase 2.79[+ or -]0.12** 2.88[+ or -]0.11***
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Mg.sup.2+] ATPase 2.63[+ or -]0.14*** 2.93[+ or -]0.14***
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)

Parameters Pepticare
 500 mg/kg 1000 mg/kg

SOD (Unit/mg protein) 3.30[+ or -]0.18 (NS) 5.07[+ or -]0.17***
Catalase ([micro]moles 7.37[+ or -]0.19 (NS) 8.28[+ or -]0.19*
 of [H.sub.2][O.sub.2]
 consumed/min/mg protein)
Reduced glutathione 2.92[+ or -]0.30*** 3.53[+ or -]0.36***
 ([micro]g of GSH/mg
 protein)
Lipid peroxidation (nmoles 4.17[+ or -]0.39*** 3.59[+ or -]0.57***
 of MDA/mg protein)
[Na.sup.+] [K.sup.+] 2.46[+ or -]0.39 (NS) 3.69[+ or -]0.40***
 ATPase ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Ca.sup.2+] ATPase 2.99[+ or -]0.10*** 3.30[+ or -]0.13***
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Mg.sup.2+] ATPase 3.30[+ or -]0.10*** 3.62[+ or -]0.17***
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)

Parameters F-value

SOD (Unit/mg protein) 26.073
Catalase ([micro]moles 6.494
 of [H.sub.2][O.sub.2]
 consumed/min/mg protein)
Reduced glutathione 14.813
 ([micro]g of GSH/mg
 protein)
Lipid peroxidation (nmoles 42.169
 of MDA/mg protein)
[Na.sup.+] [K.sup.+] 34.338
 ATPase ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Ca.sup.2+] ATPase 16.909
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)
[Mg.sup.2+] ATPase 27.149
 ([micro]moles of
 inorganic phosphorus
 liberated/min/mg
 protein)

Values are expressed as mean [+ or -] SEM.
Control group was compared with normal group.
Pepticare treated groups were compared with control group.
*p<0.05; **p<0.001; *** p<0.001; NS = non significant.

Table 3. Effect of Pepticare on the ulcer index and antioxidant
parameters in stomach of ethanol-treated rats

Parameters Normal Control

Ulcer index ([mm.sup.2]) -- 287.98[+ or -]17.79
SOD (Unit/mg protein) 5.64[+ or -]0.33 2.23[+ or -]
 0.14***
Catalase ([micro]moles of 8.27[+ or -]0.27 5.12[+ or -]
 [H.sub.2][O.sub.2] consumed/ 0.21***
 min/mg protein)
Reduced glutathione ([micro]g 3.31[+ or -]0.15 0.79[+ or -]
 of GSH/mg protein) 0.17***
Lipid peroxidation (nmoles of 3.45[+ or -]0.24 6.75[+ or -]
 MDA/mg protein) 0.72***
[Na.sup.+][K.sup.+] ATPase 5.29[+ or -]0.23 2.03[+ or -]
 ([micro]moles of inorganic 0.11***
 phosphorus liberated/min/mg
 protein)
[Ca.sup.2+] ATPase ([micro]moles 3.65[+ or -]0.12 1.50[+ or -]
 of inorganic phosphorus 0.21***
 liberated/min/mg protein)
[Mg.sup.2+] ATPase ([micro]moles 3.52[+ or -]0.36 1.49[+ or -]
 of inorganic phosphorus 0.13***
 liberated/min/mg protein)

Parameters Pepticare
 125 mg/kg 250 mg/kg

Ulcer index ([mm.sup.2]) 78.98[+ or -] 40.53[+ or -]12.7***
 14.9***
 (72.57%) (85.93%)
SOD (Unit/mg protein) 2.64[+ or -] 3.70[+ or -]0.17**
 0.14 (NS)
Catalase ([micro]moles of 5.45[+ or -] 6.21[+ or -]0.25*
 [H.sub.2][O.sub.2] consumed/ 0.19 (NS)
 min/mg protein)
Reduced glutathione ([micro]g 1.13[+ or -] 2.87[+ or -]0.40**
 of GSH/mg protein) 0.16 (NS)
Lipid peroxidation (nmoles of 5.93[+ or -] 4.52[+ or -]0.27**
 MDA/mg protein) 0.33 (NS)
[Na.sup.+][K.sup.+] ATPase 2.13[+ or -] 2.38[+ or -]0.11 (NS)
 ([micro]moles of inorganic 0.06 (NS)
 phosphorus liberated/min/mg
 protein)
[Ca.sup.2+] ATPase ([micro]moles 1.54[+ or -] 1.76[+ or -]0.14 (NS)
 of inorganic phosphorus 0.12 (NS)
 liberated/min/mg protein)
[Mg.sup.2+] ATPase ([micro]moles 1.73[+ or -] 2.52[+ or -]0.15**
 of inorganic phosphorus 0.12 (NS)
 liberated/min/mg protein)

Parameters Pepticare
 500 mg/kg 1000 mg/kg

Ulcer index ([mm.sup.2]) 16.39[+ or -]10.9*** 4.54[+ or -]
 5.6***
 (94.31%) (98.44%)
SOD (Unit/mg protein) 4.27[+ or -]0.36*** 5.37[+ or -]
 0.19***
Catalase ([micro]moles of 6.96[+ or -]0.29*** 7.80[+ or -]
 [H.sub.2][O.sub.2] consumed/ 0.20***
 min/mg protein)
Reduced glutathione ([micro]g 3.35[+ or -]0.45*** 4.56[+ or -]
 of GSH/mg protein) 0.58***
Lipid peroxidation (nmoles of 3.58[+ or -]0.28*** 3.24[+ or -]
 MDA/mg protein) 0.13***
[Na.sup.+][K.sup.+] ATPase 4.33[+ or -]0.216*** 5.49[+ or -]
 ([micro]moles of inorganic 0.08***
 phosphorus liberated/min/mg
 protein)
[Ca.sup.2+] ATPase ([micro]moles 1.90[+ or -]0.09 (NS) 2.87[+ or -]
 of inorganic phosphorus 0.05***
 liberated/min/mg protein)
[Mg.sup.2+] ATPase ([micro]moles 3.28[+ or -]0.13*** 3.46[+ or -]
 of inorganic phosphorus 0.17***
 liberated/min/mg protein)

Parameters F-value

Ulcer index ([mm.sup.2]) 149.76
SOD (Unit/mg protein) 34.032
Catalase ([micro]moles of 30.168
 [H.sub.2][O.sub.2] consumed/
 min/mg protein)
Reduced glutathione ([micro]g 20.584
 of GSH/mg protein)
Lipid peroxidation (nmoles of 14.659
 MDA/mg protein)
[Na.sup.+][K.sup.+] ATPase 142.04
 ([micro]moles of inorganic
 phosphorus liberated/min/mg
 protein)
[Ca.sup.2+] ATPase ([micro]moles 25.666
 of inorganic phosphorus
 liberated/min/mg protein)
[Mg.sup.2+] ATPase ([micro]moles 34.955
 of inorganic phosphorus
 liberated/min/mg protein)

Values are expressed as mean [+ or -] SEM.
Control group was compared with the normal group; Pepticare treated
groups were compared with control group.
*p < 0.05; **p < 0.01; ***p < 0.001; NS = non significant.
Values in parenthesis indicate the % reduction in ulcer index in
relation to the control group.

Table 4. Effect of G. glabra powder on the various gastric parameters of
pylorus-ligated and ethanol-treated rats

Gastric parameters Control G. glabra (150 mg/kg)

Ulcer index
Pylorus-ligated rats 92.75[+ or -]5.20 85.02[+ or -]5.36 (NS)
 (8.33%)
Ethanol-treated rats 287.98[+ or -]17.79 282.65[+ or -]21.22 (NS)
 (1.85%)
Volume of gastric fluid 17.28[+ or -]1.18 17.05[+ or -]0.77 (NS)
pH of gastric fluid 1.20[+ or -]0.07 1.38[+ or -]0.11 (NS)
Total acidity (mEq/1/ 127.01[+ or -]3.63 119.32[+ or -]2.56 (NS)
 100 g)

Values are expressed as mean [+ or -] SEM.
G. glabra treated groups were compared with control group. *p < 0.05;
**p < 0.01; ***p < 0.001; NS = non significant.
Values in parenthesis indicate the % reduction in ulcer index in
relation to the control group.


Acknowledgement

We are thankful to Ayur Herbals Pvt. Ltd., Baroda and UGC for providing financial assistance for successfully carrying out this work.

Received 8 July 2003; accepted 1 December 2003

References

Akhtar, M.S., Akhtar, A.H., Khan, M.A., 1992. Antiulcerogenic effects of Ocimum basilicum extracts, volatile oils and flavonoid glycosides in albino rats. Int. J. Pharmacognosy 30, 97-104.

Al-Rehaily, A.J., Al-Howiriny, T.A., Al-Sohaibani, M.O., Rafatullah, S., 2002. Gastroprotective effects of 'Amla' Emblica officinalis on in vivo test models in rats. Phytomedicine 9, 515-522.

Barry, H., 1991. Antioxidant effects a basis for drug selection. Drugs 42, 569.

Bhattacharya, A., Chatterjee, A., Ghosal, S., Bhattacharya, S.K., 1999. Antioxidant activity of active tannoid principles of Emblica officinalis (amla). Indian J. Exp. Biol. 37, 676-680.

Bonting, S.L., 1970. Presence of enzyme system in mammalian tissues. Membrane and Ion Transport. Wiley Inter Science, London 257-263.

Cheesman, K.H., 1993. Lipid peroxidation in biological systems. In: Halliwell, B., Aruoma, O.I. (Eds.), DNA and Free Radicals. Ellis Horwood, London, pp. 12-17.

Colowick, S.P., Kaplan, N.O., Packer, L., 1984. Methods in Enzymology, vol. 105. Academic Press, London 121-125.

De, B., Maiti, R.N., Joshi, V.K., Agrawal, V.K., Goel, R.K., 1997. Effect of some Sitavirya drugs on gastric secretion and ulceration. Indian J. Exp. Biol. 35, 1084-1087.

Fiske, C.H., Subbarow, Y.T., 1925. Colorimetric determination of phosphorus. J. Biol. Chem. 66, 375-400.

Halliwell, B., 1995. Antioxidant characterization: methodology and mechanism. Biochem. Pharmacol. 49, 1341-1348.

Hatano, T., Fukuda, T., Liu, Y.Z., Noro, T., Okuda, T., 1991. Phenolic constituents of licorice. Correlation of phenolic constituents and licorice specimens from various sources and inhibitory effects of licorice extracts on xanthine oxidase and monoamine oxidase. Yakugaku Zasshi 111, 311-321.

Hjerten, S., Pan, H., 1983. Purification and characterization of two forms of a low affinity [Ca.sup.2+] ATPase from erythrocyte membranes. Biochim. Biophys. Acta 728, 281-288.

Lowry, O.H., Rosenbrough, N.J., Farr, A.C., Randell, R.J., 1951. Protein measurement with folin-phenol reagent. J. Biol. Chem. 193, 265-275.

Mathew, S., Kuttan, G., 1997, Antioxidant activity of Tinospora cordifolia and its usefulness in the amelioration of cyclophosphamide induced toxicity. J. Exp. Clin. Cancer Res. 16, 407-411.

Mathur, R., Sharma, A., Dixit, V.P., Varma, M., 1996. Hypolipidaemic effect of fruit juice of E. officinalis in cholesterol-fed rabbits. J. Ethnopharmacol. 50, 61-68.

Mishra, H.P., Fridovich, I., 1972. The role of superoxide anion in the auto-oxidation of epinephrine and a simple assay for superoxide dismutase. J. Biol. Chem. 247, 3170-3175.

Mitra, A., Chakraborty, S., Auddy, B., Tripathi, P., Sen, S., Saha, A.V., Mukherjee, B., 2002. Evaluation of chemical constituents and free-radical scavenging activity of Swarnabhasma (gold ash), an ayurvedic drug. J. Ethnopharmacol. 80 (2-3), 147-153.

Mizui, T., Sato, H., Hirose, F., Doteuchi, M., 1987. Effect of antiperoxidative drugs on gastric damage induced by ethanol in rats. Life Sci. 41, 755-763.

Moron, M.S., Depierre, J.W., Mannervik, B., 1979. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim. Biophys. Acta 582, 67-78.

Ohnishi, T., Suzuki, T., Suzuki, Y., Ozawa, K., 1982. A comparative study of plasma membrane [Mg.sup.2+] ATPase activities in normal, regenerating and malignant cells. Biochim. Biophys. Acta 684, 67-74.

Pihan, G., Regillo, C., Szabo, S., 1987. Free radicals and lipid peroxidation in ethanol- or aspirin-induced gastric mucosal injury. Dig. Dis. Sci. 32, 1395-1401.

Piper, D. W., Stiel, D.D., 1986. Pathogenesis of chronic peptic ulcer, current thinking and clinical implications. Med. Prog. 2, 7-10.

Prince, P.S., Menon, V.P., 2001. Antioxidant action of Tinospora cordifolia root extract in alloxan diabetic rats. Phytother. Res. 5 (3), 213-218.

Rajeshkumar, N.V., Therese, M., Kuttan, R., 2001. Emblica officinalis fruits afford protection against experimental gastric ulcers in rats. Pharm. Biol. 39, 375-380.

Rastogi, L., Patnaik, G.K., Dikshit, M., 1998. Free radicals and antioxidant status following pylorus ligation induced gastric mucosal injury in rats. Pharmacol. Res. 38, 125-132.

Repetto, M.G., Llesuy, S.F., 2002. Antioxidant properties of natural compounds used in popular medicine for gastric ulcers. Braz. J. Med. Biol. Res. 35, 523-534.

Scartezzini, P., Speroni, E., 2000. Review on some plants of Indian traditional medicine with antioxidant activity. J. Ethnopharmacol. 71, 23-43.

Shah, Z.A., Vohora, S.B., 2002. Antioxidant/restorative effects of calcined gold preparations used in Indian systems of medicine against global and focal models of ischaemia. Pharmacol. Toxicol. 90 (5), 254-259.

Shay, H., Komarov, S.A., Fels, S.E., Meraze, D., Gruenstein, M., Siplet, H., 1945. A simple method for the uniform production of gastric ulceration in rat. Gastroenterology 5, 43-61.

Slater, T.F., Sawyer, B.C., 1971. The stimulatory effects of carbon tetrachloride and other halogenoalkanes or peroxidative reactions in rat liver fractions in vitro. Biochem. J. 123, 805-814.

Suzuki, Y., Hayashi, M., Ito, M., Yamagami, I., 1976. Anti-ulcer effects of 4'-(2-carboxyethyl) phenyl trans-4-aminomethyl cyclohexane carboxylate hydrochloride (Cetraxate) on various experimental gastric ulcers in rats. Jpn. J. Pharmacol. 26, 471-480.

Vayalil, P.K., Kuttan, G., Kuttan, R., 2002. Rasayanas: evidence for the concept of prevention of diseases. Am. J. Clin. Med. 30 (1), 155-171.

P.A. Bafna, R. Balaraman*

Department of Pharmacy, Faculty of Technology and Engineering, M.S. University of Baroda, Kalabhavan, Baroda 390 001, Gujarat, India

*Corresponding author

E-mail address: rbalaraman@satyam.net.in (R. Balaraman).
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Author:Bafna, P.A.; Balaraman, R.
Publication:Phytomedicine: International Journal of Phytotherapy & Phytopharmacology
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Date:Apr 1, 2005
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