Application Of In Vitro Cell Culture System To Screen Anti-HBV Activity Of The Experimental Study Of Chinese Medicine

Objective: Hepatitis B is a kind of important infectious disease which threat human health. The infection of HBV is extensive within the scope of the world. Currently the infectors of HBV are about 350 million. Our country is a high incidence of viral hepatitis, HBV carriers only reached 120 million, of which patients with chronic hepatitis B has more than 30 million, but has not yet been made on the fundamental breakthroughs in treatment. There are medicines such as virus inhibitory factor and nucleoside which can cause vice-reactions such as virus drug fast and active virus replication. So researching for safe and effective anti-HBV medicine has been an urgent task in the medicine world. In recent years,Chinese herbal anti-HBV study at home and abroad has become a hot spot. Chinese medicine is mainly thought it's because of the damp-heat accumulation in the liver caused by drugs in the treatment of bite to Qingrejiedu, Shugan dampness, mainly Jianpi. The study found a number of traditional Chinese medicines in anti-virus, immune regulation, there is a clear connection Baogan clinical effect.The use of the subject HBV gene-transfected human hepatoma cell HepG2. 117 single-cell models to screen herbs Scutellaria barbata, pearl grass, and Artemisia annua lucidum inhibit HBsAg, HBeAg and HBV DNA's role to provide reference for clinical treatment basis.Methods: HepG 2.117 cell culture, cell culture bottles covered with, the use of 0.25% trypsin digestion Match the concentration of 1×105 per milliliter of cells inoculated to cultivate in the 24-hole plate, each hole 1ml, buy 37℃with 5% CO2 in incubator with DMEM complete medium for liquid ,3-4d, the cell culture plate covered with the end of the experiment can be carried out. Experimental sub-group 9, including 8 drug group and a control group, each set up three parallel holes, the experimental group used drug-containing medium for fluid, blank control group with DMEM complete medium for liquid, every 24h for 1 times, respectively medication 24h, 48h, 72h after the cell supernatant collected, -20℃to preserve home test preparation. Sargent JM, etc. set up using the methyl thiazolyl tetrazolium (MTT) colorimetric assay of drugs on cell toxicity by enzyme-linked immunosorbent assay (ELISA) detected in culture supernatants of HBsAg and HBeAg levels, using fluorescent quantitative PCR assay supernatant HBV DNA level,and thereby the evaluation of anti-HBV effect of drugs.Application of statistical software SASV8.1 deal: Measurement data has T test, P <0.05 for the difference has statistical significance.Results: OD value of MTT colorimetric comparison, the experimental group and control group were no significant difference, suggesting that the selected concentrations of drugs on HepG 2.117 cell growth without toxicity. OD values of ELISA colorimetric comparison, the single Chinese medicine group, Scutellaria barbata, pearl grass, Ligustrum lucidum with the blank control group HBsAg, HBeAg compared there was a significant difference (P<0.05). Compound drug group, the Chi Lin and Chi Lin Qinggan Capsule capsule Qinggan add lamivudine with the blank control group HBsAg, HBeAg compared there was a significant difference (P<0.05). Positive control group, lamivudine and oxymatrine compared with blank control group were significantly differences (P<0.05). And artesunate group and control group no significant difference (P>0.05). HBsAg comparison, single herbs, the Scutellaria barbata at 24h and 72h when the highest inhibition rate, namely 27.98% and 33.97%, pearl grass at 48h when the highest inhibition rate for the 13.81%. Compound drugs, Chi Lin Qinggan capsules at 24h and 72h when the highest inhibition rate, namely 30.05%and 36.26%, Chi Lin Qinggan Capsule add lamivudine inhibited at 48h when the highest rate for the 24.69%. Positive control, the lamivudine at 24h, 48h and 72h when the inhibition rates are the highest, respectively, 35.75%,21.05%and 40.08%. HBeAg comparison, the single Chinese herbal medicine, Scutellaria barbata at 48h and 72h when the highest inhibition rate, namely 13.16%and 23.48%, pearl grass at 24h when the highest inhibition rate for the 5.77%; compound drugs, Chi Lin Qinggan capsules and Lamy stavudine at 24h, 48h and 72h when the inhibition rates are the highest, respectively, 14.10%,11.58%and28.34%, positive control drugs, lamivudine at 24h, 48h and 72h when the inhibition rates are the highest, respectively, 10.26%,13.68%and 28.74%. Comparison of HBV DNA, with the exception of artesunate group, the other experimental drug group and control group were significantly different (P<0.05), one of a single taste of Chinese medicine group, the highest rate of inhibition of Scutellaria barbata, for 31.36%; compound drug group, the sticks Lin Qinggan Capsule add the highest inhibitory rate of lamivudine for 42.01%, positive control drug group, the highest inhibitory rate of lamivudine for 43.42%.Conclusion: At 1mg/ml concentration, Scutellaria barbata, pearl grass, three kinds of single herbs lucidum inhibit both the role of HBV, of which the highest rate of inhibition of Scutellaria barbata, for 30.74 % , Artemisia annua without inhibiting HBV role; compound drugs, Chi Lin Qinggan Capsule add the highest inhibitory rate of lamivudine for 42.01%; positive control drugs, the role of lamivudine in inhibiting HBV best inhibitory rate of 43.42%.