Filtration Anti-hepatitis B Virus Activity Components For Natural Medicinal Plants

To filtrate the components of natural medicinal plants which have anti-hepatitis B virus activity, extracting the fractions and polysaccharide form Sarcopyramis nepalensis Wall, Phyllanthus urinaria Linn, Pholidota Lindl and evaluating the anti-HBV activity in vitro of fractions for the three natural medicinal plants Sarcopyramis nepalensis Wall, Phyllanthus urinaria Linn ans Pholidota Lindl were collected and identified in the locality. Classical hot water extracting and ethanol precipitating method were employed to isolate polysaccharide , and the content of total sugar was determined by phenol-sulfuric acid method.After the cude SWP was extracted with hot water and precipitated with ethano ,protein and pigment were also removed.HepG2.2.15 cell transfected with hepatitis B virus gene was used as cell model in vitro, cell toxicity of polysaccharide of Sarcopyramis nepalensis Wall on cultured cell line 2.2.15 was observed by MTT, and safe concentration was definite according to the TC50 and TC0. Then five concentrations of polysaccharide of Sarcopyramis nepalensis Wall and ACV were added into each well, the culture fluid was changed every 72 hours, and the supernatant of culturing fluid was collected and preserved at -20℃on the 9th day. Cell control with non-durg was set at the same time. And then the inhibition to HBsAg and HBeAg was tested by ELISA. Finally, we evaluated the anti-HBV activity in virtro of polysaccharide of Sarcopyramis nepalensis Wall according to all results above.Maximum non-toxic concentration (TC0) and half toxic concentration (TC50) of polysaccharide of Sarcopyramis nepalensis Wall on cultured cell line 2.2.15 were 3.12mg/ml and 45.41 mg/ml, and the cell toxicity was low. Half inhibitory concentrations (IC50) of polysaccharide of Sarcopyramis nepalensis Wall on HBsAg and HBeAg in cultured cell line 2.2.15 were 3.82 mg/ml and 14.60 mg/ml and , selectivity index (TI) was 11.89 and 3.11. Half inhibitory concentration (IC50) of ACV on HBsAg and HBeAg in cultured cell line 2.2.15 was 0.58 mg/ml and 0.49 mg/ml, and selectivity index (TI) was 7.86 and 9.31. The result showed that polysaccharide of Sarcopyramis nepalensis Wall could inhibit the secretion of HBsAg and HBeAg in cultured cell line 2.2.15, and had a stronger effect on HBeAg than on HBsAg, but, the inhibition effect was a little weaker than ACV.The optimum extracting procedure was simple, reasonable and reliable. The results indicated that polysaccharide of Sarcopyramis nepalensis Wall can suppress the expression of HBsAg ans HBeAg and decrease the HBV-DNA level in cultured cell line 2.2.15. The study provides therical and experimental evidences for further research of the polysaccharide Sarcopyramis nepalensis Wall, and also provides a new choice for the effective anti-HBV durg with low-toxicity.