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作 者:蒋俊俊[1] 苏锦明[1] 李彧[1] 陈晖[2] 梁冰玉[1] 黄颉刚[1] 潘沛江 刘洁[1] 赵芳凝 梁浩[2] 霍文哲[3] 叶力[1]
机构地区:[1]广西医科大学公共卫生学院,广西艾滋病防治研究重点实验室,广西南宁530021 [2]广西医科大学第一附属医院老年消化内科,广西南宁530021 [3]武汉大学动物实验中心/ABSL-3实验室,湖北武汉430000
出 处:《中华疾病控制杂志》2016年第3期244-248,共5页Chinese Journal of Disease Control & Prevention
基 金:国家自然科学基金(81271851);广西自然科学基金(2013GXNSFCB019004);广西人社厅基金(人社厅函[2013]277号)
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5μg/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,q PCR)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。Objective To investigate the effect of mycophenolic acid (MPA) on hepatitis C virus (HCV) replki- cation and the expression of autophagy-related genes treated by MPA in HCV JFH-I infected Huh7 cells. Methods Huh7 cells were infected with hepatitis C virus Japanese fulminant hepatitis -1 ( HCV JFH-I ), 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 ( LC3 B), 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 ATGT. These results suggest that MPA has a potent antiviral ability against HCV replica- tion by blocking cellular autophagy.
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