Pharmacological Evaluation Of Orthosiphon Stamineus Extract And Development Of Analgesic Meter For Arthritic Rat Model

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Date
2012-01
Authors
Yam, Mun Fei
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Abstract
Orthosiphon stamineus Benth. is a genus of herb of the family Labiatae or Lamiaceace traditionally used for treatment of many diseases such nephritis, nephrolithiasis, hydronephrosis, vesical calculi, arteriosclerosis, rheumatism, inflammation, gout and diabetes. Despite the availability of modern medications, the use of traditional medicine is growing throughout the world, indicating a need for scientific investigations into the therapeutic effects of medicinal plants and their underlying mechanisms. While no previous investigation has thoroughly reported its pharmacological activities such as anti-ulcer, anti-inflammatory, anti-pyretic, analgesic, hepatoprotective and toxicological effect of such a widely used medicinal herb, this investigation set out to further characterize the traditional claims. The O. Stamineus leaves were dried, pulverized and successively extracted with 50% methanol using maceration method. The extract was dried under reduced pressure and freeze-dried. The yield of lyophylized 50 % methanolic extract of O. Stamineus (SEOS) was found to be 6%. HPLC analysis showed that SEOS contains 0.46%, 1.12%, and 0.94% of 3’-hydroxy-5,6,7,4’- tetramethoxyflavone, sinensetin and eupatorin, respectively. In acute toxicity study, up and down method (limit dose) was adapted. A single dose of 5000 mg/kg of SEOS was given orally to 5 healthy Sprague Dawley (SD) male and female adult rats. The rats were observed for mortality and clinical signs for 3 h and then periodically for 14 days. In the subchronic toxicity study, the extract was administered orally at doses of 1250, 2500 and 5000 mg/kg per day for 28 days to female and male SD rats, respectively. The animals were sacrificed, followed by examination of their organs and blood serum. The results in the acute study showed that SEOS at a dose of 5000 mg/kg caused neither visible signs of toxicity nor mortality. All five rats survived until the end of observation period. While in subchronic toxicity, administration of the SEOS at 1250, 2500, and 5000 mg/kg for 28 days did not produce any mortality and there were no significant differences in the general condition, growth, organ weights, hematological parameters, clinical chemistry values and macroscopic appearance of the organs from the treatment groups as compared to the control group. DPPH radicals scavenging, Fe3+-induced lipid peroxidation inhibiting activities and trolox equivalent antioxidant capacity (TEAC) of SEOS were determined. The results indicated that SEOS exhibited antioxidant, lipid peroxidation inhibition and free radical scavenging activities. The hepatoprotective activity of the SEOS was studied using CCl4-induced liver toxicity in rats. The activity was assessed by monitoring liver function tests in histopathological study and measurement of alanine transaminase (ALT) and aspartate transaminase (AST). SEOS at the dose of 1000 and 500 mg/kg significantly inhibited the increase of serum ALT and AST activities and prevent the liver necrosis. Absolute ethanol-induced gastric membrane lesions model was used in antiulcer study. Oral administration of SEOS (125, 250, 500 and 1000 mg/kg) was found to significantly decrease the ulcer index (P<0.01, P<0.001, P<0.001, and P<0.001, respectively). Histological study of a section of the rat stomach also showed a marked improvement in the gastric mucosal damage in groups receiving SEOS. In order to further investigate the gastroprotective mechanism of SEOS, mucus secretion and lipid peroxidation level were estimated in vitro and ex vivo. SEOS exhibited dose-dependent stimulation of mucus secretion and inhibition of lipid peroxidation in rat gastric mucosal homogenates (both in vitro and ex vivo).
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Analgesics , Medicinal plants
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