Antiulcerogenic effects of Gymnosporia rothiana(Celastraceae) against different experimental models
AS Jain, SJ Surana
Department of Pharmacognosy, R.C.Patel College of Pharmacy, Karvand Naka, Shirpur-425405, Dist. Dhule, Maharashtra, India
|Date of Submission||07-Jan-2009|
|Date of Decision||05-Apr-2009|
|Date of Acceptance||05-May-2009|
|Date of Web Publication||16-Feb-2010|
A S Jain
Department of Pharmacognosy, R.C.Patel College of Pharmacy, Karvand Naka, Shirpur-425405, Dist. Dhule, Maharashtra
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Gymnosporia rothiana is used in Indian folk medicine as an antiulcerogenic agent. Despite of its promising use, there has been no scientific report present regarding its antiulcer activity. Therefore, this study was intended to evaluate the antiulcer property of various extracts of leaves of Gymnosporia rothiana at different dose levels in ethanol induced and indomethacin induced gastric ulcer models. It was observed that oral administration of all the extracts shows significant reduction in ulcer lesion index as well as increase in volume and pH of gastric content in both experimental models, being petroleum ether extract the most effective at dose of 250 mg/kg; it significantly reduced gastric lesion index (70.06%), in comparison to omeprazole (71.20%) and methanolic extract at a dose of 500 mg/kg (67.22%). Increased gastric mucosal defense mechanism by petroleum ether extract is probably due to its high levels of terpenoids like β amyrin, lupeol, friedelin. The present results clearly shows antiulcer effect of Gymnosporia rothiana against various irritants has been mainly due to cytoprotective effect mediated through prostaglandin and partly due to free radical scavenging activity.
Keywords: Gymnosporia rothiana , antiulcer, cytoprotective, terpenoids
|How to cite this article:|
Jain A S, Surana S J. Antiulcerogenic effects of Gymnosporia rothiana(Celastraceae) against different experimental models. Phcog Mag 2009;5, Suppl S2:100-4
|How to cite this URL:|
Jain A S, Surana S J. Antiulcerogenic effects of Gymnosporia rothiana(Celastraceae) against different experimental models. Phcog Mag [serial online] 2009 [cited 2021 Sep 23];5, Suppl S2:100-4. Available from: http://www.phcog.com/text.asp?2009/5/19/100/59792
| Introduction|| |
Peptic ulcer is the most common GIT disorder in the present day life of the industrialized and civilized world. The changing pattern of clinical evaluation and regulatory requirements for merits and demerits of drugs will be highlightened for future challenges and advances in antiulcer drug development  . In this aspect, the plant kingdom might provide a useful source of new antiulcer compounds for the development as pharmaceutical entities or alternately, as simple dietary adjuncts to existing therapies.
The celastraceae family comprises approximately 50 genus and 800 species distributed mainly tropical and subtropical regions , . The Gymnosporia genus is the largest one of the celastraceae family. Many biological activities of this genus were determined experimentally as an anticancer, antiulcerogenic, analgesic, antinociceptive, anti-inflammatory, antioxidant, antimalarial , . Gymnosporia rothiana (walp) Lawson, commonly known as Henkal, is a large armed shrub, growing western peninsular region of India  . Its leaves are used in Indian folk medicine for treatment of number of ailments including cancer, ulcer, and rheumatism  . It is also used as an anti-inflammatory, antioxidant, antiseptic, antidiarrhoeal drug. Its leaves are reported to contain n-octacosanol, β amyrin  . Despite the popular use of this species as a medicinal plant, there is no more data available about its phytochemistry and pharmacological effect. Therefore, present study has been aimed to investigate the antiulcer effects of various extracts of leaves of Gymnosporia rothiana at different dose levels.
| Materials and Methods|| |
Leaves of Gymnosporia rothiana were collected from Satpuda valley, Dist: Nandurbar, Maharashtra, India. The plant was authenticated at Department of Botany, S.S.V.P.S's College of Science, Dhule, Maharashtra. The voucher specimen has been kept at Pharmacognosy Department, R.C.Patel College of Pharmacy, Shirpur. The leaves were shed dried, ground and sieved with a 40 mesh sieve.
Preparation of extracts
About 1 kg of leaves powder was subjected to hot extraction using soxhlet extractor, successively with petroleum ether, chloroform and methanol All the extracts were filtered and concentrated under reduce pressure by using rotary flash vacuum evaporator and then dried by using vacuum dryer, giving GRPE (2.3 %), GRCL (1.8 %) and GRME (15.6 %) respectively.
Extraction of plant material for GC-MS analysis
100 gm leaves powder of Gymnosporia rothiana were extracted with ethyl acetate. After filtration, the acidic compounds were extracted out with 5% aqueous KOH (three times) followed by the extraction of the basic compounds with 5% aqueous HCl (three times). The organic fraction, which contain the neutral compounds, was washed with water to pH- 7 and concentrated in rotary vacuum evaporator to 50 ml. Suspended particle were removed by centrifuging the concentrated extract for 10 min at 6000 rpm. Then solvent was evaporated to dryness, giving a residue, which was dissolved in chloroform for GC-MS analysis.
GC-MS analysis was performed on a Hewlett-Packard 5890 gas chromatograph, with a split injector (1:50) at 280 °C and a Hewlett-Packard 5970 mass selective detector (MSD), with the GC-MS interface temperature at 280 °C. The injection volume was 2μl. Hydrogen was employed as carrier gas, at a pressure of 60 kPa. A HP-1 25m Χ 0.25 mm Χ 0.33 μm methylpolysiloxane cross-linked capillary column was employed with temperature programming from 100 °C (held for 2 min) to 280 °C (held for 30 min) at a ratio of 4 °C/min.
The study was performed with male wistar rats (150-200 g), housed in standard environmental conditions (25 ± 3°C and humidity 60 ± 5%) under a 12h dark: 12h light cycle. During maintenance animals received a diet of food pellets (Amrut Labs, Pune) and water ad libitum. Before the experiments, the animals were deprived of food for 24hrs. Experimental protocol were designed to meet the "Guidelines of animal experimentation", approved by the ethical committee of the institute.
Antiulcer activity was evaluated using two different assay models for induction of gastric lesions: NSAID- induced (indomethacin) and absolute ethanol- induced gastric lesions  . For sake of comparison, animals were treated with omeprazole and cimetidine, depending on the experiment. At the end of each experiment, the animals were sacrificed by cervical dislocation; the stomach was removed and its gastric content was collected. Then stomach opened along the greater curvature and fixed between two glass plates. Free acidity, total acidity, volume and pH of gastric content were calculated as per standard methods. Ulcer lesion index calculated by severity of gastric mucosal lesion  , graded as follows: 1) Loss of normal morphology, discoloration of mucosa, mucosal damage, hemorrhagic streaks (1 point each) 2) petechial point (< 10, 2 points, =10, 3 points) and 3) No. / Size of the ulcers (number of ulcer until 1mm Χ 2 points, larger than 1 mm Χ 3 points) 4) perforated ulcer (number of ulcer Χ 4 points). The percentage determination is as follows:
Ethanol induced gastric lesion
After 24 h fasting, the rats were divided into eight groups of six animals each. The group I served as a normal control, given 1% CMC in water (5 ml/kg, p.o), Group II was treated orally with omeprazole (30 mg/kg), Group III, V, and VII orally received 250 mg/kg of petroleum ether, chloroform, and methanol extract respectively while Group IV, VI, and VIII orally received 500 mg/kg of petroleum ether, chloroform, and methanol extract respectively. After 45 min, ulceration was induced by oral administration of 1 ml of absolute ethanol. Animals were sacrificed after 1h following the administration of absolute ethanol , .
Indomethacin induced gastric lesion
After 24 h fasting, the rats were divided into eight groups of six animals each. The group I served as a normal control, given 1% CMC in water (5 ml/kg, p.o), Group II was treated orally with cimetidine (100 mg/kg). Remaining groups (group III - VIII) were treated as previously described. After 45 min, ulceration was induced by oral administration of 100 mg/kg of indomethacin. Animals were sacrificed after 1h following the administration of indomethacin  .
All the results are reported as mean ± SEM. The statistical analysis was carried out using one-way ANOVA followed by Dennett's multiple comparison and p value < 0.05 were considered statistically significant.
| Results|| |
Phytochemical investigations of petroleum ether extract and chloroform extract revealed the presence of terpenes and steroids while methanol extract shows the presence of condensed tannins, flavanoids, saponin glycosides, alkaloids, proteins & amino acids.
The result of GC-MS analysis of Gymnosporia rothiana were given in [Table 1], which shows GC-MS experimental data, retention time (RT), and mass fragment of compounds for terpenoids and steroids of Gymnosporia rothiana. Individual compound were identified from RT, mass data and by comparison of the data of standard compounds with those of in the literature. Ten compounds viz. Friedelin, Hop-22(29)-en-3.beta-ol, lupeol, β amyrin, campesterol, ergosterol, sitosterol, squalene, phytol, palmitic acid were identified.
Evaluation of antiulcer activity
All the extracts of Gymnosporia rothiana showed a dose dependant gastroprotective effect against ethanol induced [Table 2] and [Table 3], and indomethacin induced gastric ulcer [Table 4] and [Table 5]. Petroleum ether extract at dose of 250 mg /kg significantly protected mucosal damage (70.06%), in comparison to omeprazole (71.20%) and methanolic extract at a dose of 500 mg/kg (67.22%).
| Discussion|| |
Peptic ulcer occurs when there is an imbalance between the damaging effects of gastric acid and pepsin and the defense mechanisms, which protects gastric mucosa from these substances. Therefore, effective drug against peptic ulcer are those which basically act either by reducing the aggressive factors or by stimulating mucosal defense. The genesis of ethanol induced gastric ulcer is associated with disturbances in gastric secretion, damage to gastric mucosa, alteration in permeability, gastric mucus depletion and also with free radical production, leads to increased lipid peroxidation which in turn causes damage to cell and cell membrane , . While non steroidal anti-inflammatory drugs like indomethacin are known to induce ulcer by inhibiting prostaglandin synthetase through cyclooxygenase pathway. In the stomach, prostaglandin plays a vital protective role, stimulating the secretion of bicarbonate and mucus, regulating mucosal cell turnover and repair. Thus the suppression of prostaglandin synthesis by NSAIDS results in increased susceptibility to mucosal injury and gastric ulcer  .
Phytochemical investigations revealed the presence of terpenes and steroids like Friedelin, Hop-22(29)-en-3.beta-ol, lupeol, β amyrin, campesterol, ergosterol, sitosterol, squalene and phytol. Methanol extract shows the presence of condensed tannins, flavanoids, saponin glycosides, alkaloids, proteins & amino acids. A review on antiulcer drugs of plant origin shows that Triterpenes like β amyrin, lupeol acetate, ursolic acid, glycerrhetinic acid and sterols like β sitosterol exert their antiulcer effect by strengthening defensive factors such as stimulation of mucus synthesis or maintenance of prostaglandin contents of gastric mucosa at high levels  . In addition, these compounds act as antioxidant which protects gastric mucosa against oxidative damage  . Furthermore extracts containing flavanoids also shows antisecretory, cytoprotective and antioxidant activity  .
It was observed that oral administration of petroleum ether, chloroform and methanol extracts of Gymnosporia rothiana produces significant reduction in ulcer lesion index as well as increase in volume and pH of gastric content in both ethanol and indomethacin induced gastric ulcer models, being petroleum ether extract the most effective, it increased the gastric mucosal defense mechanism due to its high levels of terpenoids.
| Conclusion|| |
Present results clearly shows antiulcer effect of extracts of Gymnosporia rothiana against various irritants has been mainly due to cytoprotective effect mediated through prostaglandin and partly due to free radical scavenging activity. Thus the study provides for the first time evidence that showed antiulcer effect of Gymnosporia rothiana which correlate with its folklore claim as an antiulcer drug.
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[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]
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