The effect of mycophenolic acid on expression of autophagy-related genes in hepatocytes
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摘要: 目的 比较日本爆发性丙型肝炎病毒1型(hepatitis C virus Japanese fulminant hepatitis-1, HCV JFH-1)感染的Huh7细胞中细胞自噬因子在霉酚酸(mycophenolic acid,MPA)处理组与对照组的表达差异, 探讨MPA对肝细胞自噬相关因子的调节影响。方法 用HCV JFH-1感染Huh7细胞,用5 μg/ml的霉酚酸处理细胞24 h,用实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction, qPCR)和蛋白免疫印迹法测定细胞内HCV、微管相关蛋白1轻链3(microtubule-associated-protein 1 light chain, MAP1-LC3B, 简写LC3B)、自噬相关因子3(autophagy-related gene 3, ATG3)和自噬相关因子7(autophagy-related gene 7, ATG7)的信使核糖核酸(messenger ribonucleic acid,mRNA)和蛋白相对表达量,比较上述因子在处理组和对照组之间的差异。结果 在核糖核酸(ribonucleic acid,RNA)和蛋白水平上,MPA可以降低HCV在Huh7细胞中的表达并且降低自噬标记物LC3B mRNA和LC3B-II的蛋白质表达(均有P<0.001);MPA可以使Huh7细胞中的自噬相关因子ATG3和ATG7的mRNA和蛋白相对表达量下降(P<0.001;P=0.003; P=0.010;P=0.002)。结论 MPA可以抑制HCV JFH-1在Huh7细胞中的复制和细胞自噬,并且可以下调自噬因子ATG3和ATG7的表达,推测MPA可能通过调节自噬关键因子表达来抑制细胞自噬从而拮抗HCV的复制。Abstract: Objective To investigate the effect of mycophenolic acid (MPA) on hepatitis C virus (HCV) replkication and the expression of autophagy-related genes treated by MPA in HCV JFH-1 infected Huh7 cells. Methods Huh7 cells were infected with hepatitis C virus Japanese fulminant hepatitis -1(HCV JFH-1), and then treated with 5μg/ml MPA for 24h. The treated cells were collected to detect the expression of HCV, microtubule-associated protein 1 light chain(LC3B), autophagy-related gene 3(ATG3) and autophagy-related gene 7(ATG7) at messenger ribonucleic acid (mRNA) and protein levels by real-time quantitative PCR(qPCR) and Western blot assay respectively. Results MPA treatment decreased the expression of HCV and LC3B mRNA and LC3B-II protein (P<0.001, compared with control group), and reduced the expression of ATG3 and ATG7 at mRNA and protein levels (P<0.001, P=0.003; P=0.010, P=0.002 compared with control group). Conclusions MPA treatment could inhibit HCV replication and block autopahgy by down-regulating the expression of ATG3 and ATG7. These results suggest that MPA has a potent antiviral ability against HCV replication by blocking cellular autophagy.
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Key words:
- Mycophenolic acid /
- Autophagy /
- Hepatitis C
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