Penilaian hepatoprotektif ekstrak kloroform phyl/anthus pulcher wall. ex mull. arg. dan penyisihan flavonoid daripada ekstrak metanolnya berdasarkan aktiviti penyingkiran radikal bebas
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Date
2007
Authors
Suh In, Loh
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Abstract
Preliminary study has shown a strong cytotoxic activity of chloroform extract of
Phyllanthus pulcher against in vitro human hepatocellular carcinoma (HepG2) cell line.
Therefore, further investigation on the hepatoprotective effect of the chloroform extract
has been carried out on the in vivo CCI4-induced liver damage in mice. The acute and
sub-acute toxicological tests were conducted in order to determine the doses for the
hepatoprotective study. From the acute toxicity test, the extract was considered having
moderately toxicity with a slightly low LD50 (119.8 mg/kg body weight) and the result
suggested the use of lower but repeated doses in sub-acute toxicity test which was
conducted for 14 days. No significant adverse effects observed from the animals in the
test except the decrease of body weight at end of the test on female mice in groups
dosed 5 and 10 mg/kg bw and the increase of relative weight of lungs on male mice in
group dosed 2.5 mg/kg bw. On the other hand, the chloroform extract has shown very
well protective effects on the liver injury induced by CCI4 . Pre-treated groups of extract
have significantly decreased the induced level of liver enzymes (AL T and AST) and it is
supported by the histological results. Nevertheless, the post-treated groups showed
better effect by reducing enzymes level to normal. The hepatoprotection of posttreated
groups showed an equivalence effectiveness to silymarin which served as
positive control. Meanwhile, an evaluation of in vitro antioxidant activity has also been
carried out as another part of this study. The free radical DPPH was used to evaluate
for the antioxidant property of five extracts of P. pu/cher. Methanolic extract has been
chosen for the bioassay-guided fractionation because it possessed the best
scavenging activity. The result was also supported by the observation of non-
spectrophotometry study. Fraction F5 (with 86 % of scavenging activity at the final
concentration of 125 IJg/ml) from the first fractionation has been separated to get bands
of F5-6, F5-7 and F5-8 which possessed a high score of scavenging percentage (82-
85 % at the same final concentration). Mixture of those bands was then being
hydrolyzed to obtain the aglycone. The aglycones were fractionated and their
scavenging activities were assessed. Subtraction of F5-CAII was eventually identified
tentatively as kaempferol and the identification was supported by the results from UVVisible
spectrophotometry, LC-MS and TLC. ECso of methanol extract, fraction F5-C,
subtraction F5-CAII and other flavonols (kaempferol, quercetin and myricetin) were
determined. EC50 of subtraction and kaempferol or, and fraction F5-C are not differed
from each other (p>0.05) respectively, and have higher value than that of quercetin and
myricetin. EC50 of methanol extract is the highest in this study.
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Keywords
Hepatoprotektif ekstrak kloroform , Phyllanthus pulcher wall. ex mull. arg. , Penyingkiran radikal bebas