Lupine Publishers| Archives of Nanomedicine Open Access Journal (ANOAJ)
Abstract
Objective: Leea indica is a well known plant with numerous pharmacological activities owing to the presence of the active constituents. Inspite of numerous therapeutic uses, efficacy of the plant in treating ulcers is not yet evaluated scientifically. Hence an attempt has been made to evaluate the Antiulcer activity of Leea indica.
Methods: The effect of Leea indica methanolic extract (LIME) on gastric ulcer in pylorus ligation-induced and aspirin induced models was studied by employing LIME at 200 mg/kg, 400 mg/kg. In both the models Ranitidine (40mg/kg) was employed as the standard. Depending on the model, parameters evaluated were total acidity, free acidity, volume and pH of gastric fluid, ulcer score and percent inhibition of ulcer index.
Results: Data were analyzed following graph pad instat version 3.0 followed by Dunnets multiple comparision test. LIME (400mg/kg) significantly (P<0.01) reduced gastric ulcer index in pylorus ligation-induced and Aspirin induced ulcer models comparable to that of Standard drug. Histopathological studies confirmed that LIME (400mg/kg) possess anti ulcer activity in both models. The elicited activity might be owing to the existence of secondary metabolites such as flavonoids, tannins, and saponins.
Conclusion: Obtained results elucidate the Antiulcer activity of Leea indica methanolic extract. Further investigations on isolation of specific phyto chemicals and elucidating mechanisms of action are needed.
Keywords:Antiulcer, Pylorus Ligation Model, Aspirin Induced Model, Histopathology, Leea Indica, Peptic Ulceration, Vitaceae, Anti Hyper Lipidaemic, Antitumor, Hydrocarbons
Mini Review
Peptic ulceration is one of the common modern age epidemics affecting nearly 10% of world population [1]. Even though numerous potent anti-ulcer drugs are marketed, most of them were associated with toxicities, thus emphasizing the essentiality in search for new alternatives. Approximately 80% of the world’s population trust on herbal-derived medicines reinforcing herbs was advantageous sources for new drugs. Leea indica a traditional Chinese medicine found to be distributed in India, Srilanka, Bangladesh, Burma, Nepal, Thailand, Laos, Camboida, China, Vietnam, New Guinea, Malasia, Solomon islands, North Australia, Santa Cruz island, New Hebrides and Fiji [2-4]. Leea indica, an evergreen large shrub of family vitaceae [5,6] grows up to 8mt. Leaflets are ovate-lanceolate with crenate to serrate margins. Leaves are 1-3 pinnate bearing 7 leaflets, with petioles 7-20cm long. Fruits are purplish black, bearing six seeds with 1 cm in diameter. Flowers are greenish white with 5mm across. Stems are glabrous to pubescent.
Leea indica leaves were scientifically evidenced for wide pharmacological activities like antitumor [7], antiviral [8], analgesic [9], sedative and anxiolytic [10], Nitric oxide inhibitory [11], phosphodiestrase inhibitory [12], antidiabetic [13]. Anti hyper lipidaemic [14]. Leaf decoction is employed in pregnancy and delivery for birth control and body pain [4,15]. Dried leaves ingested were effective against cancer [16]. Roasted leaves relieve vertigo. Leaves (31.4%) constitute to be the most widely used parts in medicine preparation then fruits and roots (16.05%) [17]. Therefore the current investigation was designed to screen the leaves for the antiulcer activity. Twenty three chemical constituents were detected in Leea indica leaves by Gc-Ms analysis, Spectroscopic techniques, and Co-TLC. Compounds identified were eleven hydrocarbons, palmitic acid, pthalic acid, 1-eicosanol,farnesol, soalnesol, gallic acid, three pthalic acid esters, lupeol, β-sitosterol and finally ursolic acid [18]. Though the plant was reported with numerous chemical constituents antiulcer activity was not yet screened. Hence the present study was taken up to investigate antiulcer activity of Leea indica.
Materials and Methods
Chemicals
Ranitidine(Lee Pharmaceuticals, Hyderabad.), Aspirin (Shalg pharmaceuticals, Goregaon, Mumbai.)
Plant
(Figure 1) Leea indica leaves were procured from Karthikavanam forest, Dhulapally, Hyderabad. These were authenticated by Dr. Madhavachetty, Professor, Sri Venkateshwara University, Tirupathi. The plant was placed in the college herbarium with a vocher specimen no. 438.
Figure 1: Leea indica leaves.
Animals
Male Wistar albino rats (200–250g) were purchased at Albino Research Center, Bachupally, Hyderabad. They were accomodatedin polypropylene cages with standard diet and water ad libitum. Experiment was designed as per ethical norms of CPCESA and IAEC Malla Reddy Institute of Pharmaceutical Sciences (1662/PO/a/12/ CPCSEA).
Extraction
Fresh leaves were shade dried, coarsely powdered and sieved with sieve no.40. Powdered material (500gm) was packed in soxhlet apparatus and subjected to extraction with methanol and water for about 48h. Solvents were evaporated under reduced pressure by rotary vacuum evaporator. Obtained methanolic extract (MELI) and aqueous extracts of Leea indica (AQLI) were subjected to phyto chemical screening and acute toxicity study [19].
Acute Toxicity Study
This study was performed as per OECD guideline 425 using limit test dose 2000mg/kg [20]. Rats were fasted overnight and a limit dose of 2000mg/kg was administered and observed for autonomic profiles (defecation and urination), behavioural profile (alertness, restlessness, irritability and fearfulness), neurologic profile (spontaneous activity, reactivity, touch response, pain response and gait), physical states such as lacrimation, loss of appetite, tremors, hair erection, salivation, diarrhoea and for morbidity or mortality continuously for 2 h, periodically during the first 24 h and daily thereafter, for a total of 14d.
Antiulcer Activity
Pylorus Ligation Method: Male Wistar albino rats were segregated into four groups comprising five each [21]. Group I negative control, received distilled water, Group II positive control, received ranitidine 50mg/kg; Group III LIME (200mg/kg, p.o); Group IV LIME (400mg/ kg, p.o). Plant extracts were administered for 10 d. At the end of experimental period rats were fasted overnight with water ad libitum. Standard, Ranitidine (40mg/kg) and LIME (200, 400mg/ kg) were administered. Under light ether anaesthesia, the abdomen was cut opened by incision below the xiphoid process. The Pylorus end of stomach was slightly lifted up and ligated avoiding damage to blood vessels. After completing ligation stomach was replaced and the abdominal wall is sutured and closed. Rats were deprived of water during post operative period for 3 to 4h [22]. Four hours later, stomach was dissected out. Stomach contents were drained into tubes and were subjected to centrifugation at 2000 r/m for 10 min. Supernatant obtained was analysed for gastric volume, pH, free acidity and total acidity. The stomach was incised along the greater curvature, and observed for ulcers and the ulcer index (UI) was calculated. Ulcers were scored and severity was assessed microscopically (10 X) employing hand lens (10 X). The scores were given as mentioned.
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