MKS001等四种药物体外抑制HBV的实验研究

发布时间：2018-05-05 13:42

本文选题：药物 + HBV　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:研究MKS001、XSJ002、MGS003、NKD004四种药物体外对乙型肝炎病毒的抑制作用,为乙型肝炎病毒的治疗及治疗机制提供新的探索思路和途径。方法:HepG2.2.15细胞是由HBV全基因组转染人肝癌细胞株得到的可以在体外稳定的、高水平表达HBsAg、HBeAg及完整的HBV颗粒的细胞株。通过体外培养HepG2.2.15细胞,向细胞培养液中分别加入上述四种不同浓度的药物,观察每种药物在不同浓度下的细胞病变程度,并利用CCK-8法检测药物对细胞的毒性作用。在最大无毒性浓度下,将上述四种药物再配置成不同浓度,然后分别加入HepG2.2.15细胞中培养72h后收集细胞培养液,用ELISA法检测细胞培养液中HBsAg和HBeAg,用荧光定量PCR法检测细胞培养液中HBVDNA载量。采用SPSS16.0软件系统中的t检验进行统计学分析,计量资料以均数±标准差(X±S)表示。P0.05具有统计学意义。结果:MKS001、XSJ002、MGS003在体外均有抑制HBV的作用。MKS001的最大无毒性浓度(TC0)为1umol/L,此时对HepG2.2.15细胞体外分泌的HBsAg的抑制率为59.96%(P0.01),对HBe Ag的抑制率为35.17%(P0.01),对HBVDNA复制的抑制率为60.00%(P0.01);XSJ002的最大无毒性浓度(TC0)为0.01umol/L,此时对HepG2.2.15细胞体外分泌的HBsAg的抑制率为27.12%(P0.01),对HBeAg的抑制率为33.93%(P0.01),对HBVDNA复制的抑制率为46.47%(P0.01);MGS003的最大无毒性浓度(TC0)为0.01umol/L,此时对HepG2.2.15细胞体外分泌的HBsAg的抑制率为25.46%(P0.01),对HBe Ag的抑制率为31.43%(P0.01),对HBVDNA复制的抑制率为29.17%(P0.01);NKD004则在所选的药物浓度范围内均未发现体外对HBsAg、HBe Ag和HBVDNA有抑制性。结论:1、MKS001、XSJ002、MGS003体外均有抑制HBV作用,NKD004体外未发现抗HBV作用。2、MKS001、XSJ002、MGS003三种药物对HBV的抑制强度均随药物浓度的增大而增加,体现出明显的药物剂量依赖性。
[Abstract]:Objective: to study the inhibitory effect of four drugs (MKS001, XSJ002, MGS003, NKD004) on hepatitis B virus in vitro, and to provide new ideas and approaches for the treatment and mechanism of hepatitis B virus. Methods: human HepG2.2.15 cells were transfected into human hepatoma cell line by HBV genome, which could express HBsAg HBeAg and complete HBV granules at high level in vitro. HepG2.2.15 cells were cultured in vitro and four different concentrations of drugs were added to the cell culture medium to observe the degree of cytopathic effect of each drug at different concentrations. The cytotoxicity of the drugs to cells was detected by CCK-8 method. At the maximum nontoxic concentration, the above four drugs were reconfigured into different concentrations, then cultured in HepG2.2.15 cells for 72 hours and then collected the cell culture medium. HBsAg and HBeAg in cell culture medium were detected by ELISA assay and HBVDNA load in cell culture medium was detected by fluorescence quantitative PCR method. T test in SPSS16.0 software system was used for statistical analysis. The measurement data were expressed as mean 卤standard deviation (X 卤S). P05 was statistically significant. Results: MKS001XSJ002MGS003 inhibited HBV in vitro. The maximum nontoxic concentration (TC0) of MKS001 was 1 umolrL. The inhibition rate of HBsAg secreted by HepG2.2.15 cells in vitro was 59.96 (P0.01N), the inhibition rate of HBe Ag was 35.17171.The inhibition rate of HBVDNA replication was 60.005% P0.01XSJ002. TC0 was 0.01umol/ L, the inhibition rate of HBsAg secreted by HepG2.2.15 cells in vitro was 27.12 and that of HBeAg was 33.93mg / L, the inhibition rate of HBVDNA replication was 46.47mol / L, the maximum nontoxic concentration TC0 of HBVDNA was 0.01umolL, the inhibition rate of HBsAg secreted by HepG2.2.15 cells in vitro was 25.464mg / r, and the inhibition rate on HepG2.2.15 cells in vitro was 25.464mg / r. The inhibition rate of HBe Ag was 31.43mg / kg, and the inhibition rate on HBVDNA replication was 29.17 and 29.17 respectively. However, no inhibition of HBe Ag and HBVDNA in vitro was found in the selected drug concentration range. Conclusion the inhibitory effect of MKS001, XSJ002, MGS003 on HBV in vitro was not found in NKD004. The inhibitory intensity of three drugs against HBV was increased with the increase of drug concentration, which showed a significant dose-dependent manner, and the inhibitory effect of MKS001XSJ002MGS003 on HBV was not found in vitro.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.62

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