本文選題：藥物 + HBV��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究MKS001、XSJ002、MGS003、NKD004四種藥物體外對(duì)乙型肝炎病毒的抑制作用,為乙型肝炎病毒的治療及治療機(jī)制提供新的探索思路和途徑。方法:HepG2.2.15細(xì)胞是由HBV全基因組轉(zhuǎn)染人肝癌細(xì)胞株得到的可以在體外穩(wěn)定的、高水平表達(dá)HBsAg、HBeAg及完整的HBV顆粒的細(xì)胞株。通過(guò)體外培養(yǎng)HepG2.2.15細(xì)胞,向細(xì)胞培養(yǎng)液中分別加入上述四種不同濃度的藥物,觀察每種藥物在不同濃度下的細(xì)胞病變程度,并利用CCK-8法檢測(cè)藥物對(duì)細(xì)胞的毒性作用。在最大無(wú)毒性濃度下,將上述四種藥物再配置成不同濃度,然后分別加入HepG2.2.15細(xì)胞中培養(yǎng)72h后收集細(xì)胞培養(yǎng)液,用ELISA法檢測(cè)細(xì)胞培養(yǎng)液中HBsAg和HBeAg,用熒光定量PCR法檢測(cè)細(xì)胞培養(yǎng)液中HBVDNA載量。采用SPSS16.0軟件系統(tǒng)中的t檢驗(yàn)進(jìn)行統(tǒng)計(jì)學(xué)分析,計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差(X±S)表示。P0.05具有統(tǒng)計(jì)學(xué)意義。結(jié)果:MKS001、XSJ002、MGS003在體外均有抑制HBV的作用。MKS001的最大無(wú)毒性濃度(TC0)為1umol/L,此時(shí)對(duì)HepG2.2.15細(xì)胞體外分泌的HBsAg的抑制率為59.96%(P0.01),對(duì)HBe Ag的抑制率為35.17%(P0.01),對(duì)HBVDNA復(fù)制的抑制率為60.00%(P0.01);XSJ002的最大無(wú)毒性濃度(TC0)為0.01umol/L,此時(shí)對(duì)HepG2.2.15細(xì)胞體外分泌的HBsAg的抑制率為27.12%(P0.01),對(duì)HBeAg的抑制率為33.93%(P0.01),對(duì)HBVDNA復(fù)制的抑制率為46.47%(P0.01);MGS003的最大無(wú)毒性濃度(TC0)為0.01umol/L,此時(shí)對(duì)HepG2.2.15細(xì)胞體外分泌的HBsAg的抑制率為25.46%(P0.01),對(duì)HBe Ag的抑制率為31.43%(P0.01),對(duì)HBVDNA復(fù)制的抑制率為29.17%(P0.01);NKD004則在所選的藥物濃度范圍內(nèi)均未發(fā)現(xiàn)體外對(duì)HBsAg、HBe Ag和HBVDNA有抑制性。結(jié)論:1、MKS001、XSJ002、MGS003體外均有抑制HBV作用,NKD004體外未發(fā)現(xiàn)抗HBV作用。2、MKS001、XSJ002、MGS003三種藥物對(duì)HBV的抑制強(qiáng)度均隨藥物濃度的增大而增加,體現(xiàn)出明顯的藥物劑量依賴性。
[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 鹵standard deviation (X 鹵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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R512.62

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