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《Journal of Clinical Rehabilitative Tissue Engineering Research》 2007-38
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Extraction and primary isolation of hepatoprotective active ingredients on Phyllanthus urinaria L.and carbon tetrachloride-injured models of primary hepatocytes in rats

Fan S,Zuo JP,Yi C,Peng GP,Yang H,Shi GR,Rao LQ. 1College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha 410128,Hunan Province,China; 2Hunan Environment-Biological Polytechnic,Hengyang 421005,Hunan Province,China  
AIM:To extract and primarily isolate the active components from Phyllanthus urinaria L. and investigate the hepatoprotective potential of the monomer component in the pharmacological model of primary culture hepatocytes in rat following chemical injury induced by carbon tetrachloride. Hepatoprotective potential is tested by method of Methylthiazoletrazolium (MTT) colorimetry. METHODS: From May to December in 2006,an experiment was accomplished in the Key Laboratory of Traditional Chinese Medicine Development and Estimation in Zhejiang University Pharmaceutical College.①Newborn rat was used to prepare the pharmacological model that the hepatocytes were injured by carbon tetrachloride.②Preparation of Phyllanthus urinaria L. extracts: The active component of Phyllanthus urinaria L. was extracted by back-flow adopting water and alcohol,in which the volume fraction was 0.95.③Primary isolation of the active component: The extraction was performed in proper order with the petroleum ether,chloroform,ethyl acetate and n-butanol; The substep elution applied AB-8 big-hole for resin adsorptions,and adopted the alcohol of 0.1-0.9 volume fraction.④Using MTT colorimetry,the hepatoprotective potential of back-flow extract and elution by AB-8 big-hole for hepatocytes in rat injured by carbon tetrachloride was evaluated through the protection rate and the proliferation index. RESULTS: ①The extract from n-butanol solvent had higher protection rate and the proliferation index than positive controls and other extracts (P 0.05-0.01).②In the extract of n-butanol,the elution by 0.1,0.3,0.5 volume fractions of alcohol had hepatoprotective potential,and that by 0.1 volume fraction of alcohol was the best,in which hepatoprotective potential was remarkably better than positive controls and other fractions (P 0.01). At the concentration of 200 g/L,the protective rate of elution by 0.1 volume fraction of alcohol was 64.43%,and the proliferation index was 2.952. CONCLUSION: The effective location of Phyllanthus urinaria L. to protect livers is AB-8 big-hole resin adsorptions at 0.1 volume fraction of alcohol in n-butanol extract. But the effective monomer of Phyllanthus urinaria L. does not ascertain and remains to be proved in the further experiment.
【Fund】: 湖南省教育厅青年基金(04B018)~~
【CateGory Index】: R-332;R284
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