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Synergistic effect of a novel oxymatrine-baicalin combination against hepatitis B virus replication α smooth muscle actin expression and type I collagen synthesis in vitro

机译:新型氧化苦参碱-黄ical苷组合物对乙型肝炎病毒复制α平滑肌肌动蛋白表达和I型胶原合成的协同作用

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摘要

AIM: To study the effect of oxymatrine-baicalin combination (OB) against HBV replication in 2.2.15 cells and α smooth muscle actin (α SMA) expression, type I, collagen synthesis in HSC-T6 cells.METHODS: The 2.2.15 cells and HSC-T6 cells were cultured and treated respectively. HBsAg and HBeAg in the culture supernatants were detected by ELISA and HBV DNA levels were determined by fluorescence quantitative PCR. Total RNA was extracted from HSC-T6 cells and reverse transcribed into cDNA. The cDNAs were amplified by PCR and the quantities were expressed in proportion to β actin. The total cellular proteins extracted from HSC-T6 cells were separated by electrophoresis. Resolved proteins were electrophoretically transferred to nitrocellulose membrane. Protein bands were revealed and the quantities were corrected by β actin.RESULTS: In the 2.2.15 cell culture system, the inhibitory rate against secretion of HBsAg and HBeAg in the OB group was significantly stronger than that in the oxymatrine group (HBsAg, P = 0.043; HBeAg, P = 0.026; respectively); HBV DNA level in the OB group was significantly lower than that in the oxymatrine group (P = 0.041). In HSC-T6 cells the mRNA and protein expression levels of α SMA in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P = 0.013; protein, P = 0.042; respectively); The mRNA and protein expression levels of type I collagen in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P < 0.01; protein, P < 0.01; respectively).CONCLUSION: OB combination has a better effect against HBV replication in 2.2.15 cells and is more effective against α SMA expression and typeI collagen synthesis in HSC-T6 cells than oxymatrine in vitro.
机译:目的:研究氧化苦参碱-黄ical苷(OB)对2.2.15细胞中HBV复制和α平滑肌肌动蛋白(αSMA)表达,HSC-T6细胞Ⅰ型胶原合成的影响。方法:2.2.15培养细胞和处理HSC-T6细胞。通过ELISA检测培养上清液中的HBsAg和HBeAg,并通过荧光定量PCR测定HBV DNA水平。从HSC-T6细胞中提取总RNA,然后反转录为cDNA。通过PCR扩增cDNA,其数量与β肌动蛋白成比例表达。通过电泳分离从HSC-T6细胞提取的总细胞蛋白。将溶解的蛋白质电泳转移到硝酸纤维素膜上。结果:在2.2.15细胞培养系统中,OB组对HBsAg和HBeAg分泌的抑制率明显强于氧化苦参碱组(HBsAg,P = 0.043; HBeAg,P = 0.026;)。 OB组的HBV DNA水平显着低于氧化苦参碱组(P = 0.041)。在HSC-T6细胞中,与氧化苦参碱组相比,OB组中αSMA的mRNA和蛋白表达水平显着降低(mRNA,P = 0.013;蛋白质,P = 0.042;)。与氧化苦参碱组相比,OB组Ⅰ型胶原蛋白的mRNA和蛋白表达水平显着降低(mRNA,P <0.01;蛋白质,P <0.01;结论):OB联合抗氧化效果更好。 HBV在2.2.15细胞中复制,并且比体外氧化苦参碱对HSC-T6细胞中的αSMA表达和I型胶原合成更有效。

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