氧化苦参碱对miR-122低表达、感染HBV肝细胞的抗病毒作用观察  

Antiviral effect of oxymatrine on low expression of miR-122 and HBV infection of hepatocytes

作　　者：张轩[1] 王淏[2] ZHANG Xuan;WANG Hao(Guangdong Provincial TCM Hospital,Guangzhou 510120,China)

机构地区：[1]广东省中医院/广州中医药大学第二附属医院,广州510120 [2]广州市血液中心

出　　处：《山东医药》2018年第23期26-28,共3页Shandong Medical Journal

基　　金：广东省中医药局基金资助项目(20152140)

摘　　要：目的观察氧化苦参碱(OMT)对miR-122低表达、感染HBV肝细胞的抗病毒作用。方法体外培养Hep G2.2.15细胞,分为A、B、C、D、E、F组。细胞接种培养板24 h后将A、B、C组更换为无血清DMEM培养基,D、E、F组更换为配制好的含4 mg/m L OMT的无血清DMEM培养基。同时A、D组加入Lipofectamine2000;B、E组加入Lipofectamine2000和miR-122抑制剂hsa-miR-122-5p inhibitor;C、F组加入Lipofectamine2000和miR-122抑制剂阴性对照inhibitor Negative Control#22。48 h后收集各组细胞,采用荧光定量PCR法检测miR-122。于转染后48 h收集各组培养液上清和细胞,采用电化学发光法检测培养液上清中的HBsAg和HBeAg,采用荧光定量PCR法检测细胞中的HBV DNA。结果 A、B、C、D、E、F组miR-122相对表达量分别为1.00±0.26、0.72±0.15、1.04±0.28、3.09±0.94、1.27±0.59、3.00±0.62。B组miR-122相对表达量小于A、C组(P均<0.05);E组miR-122相对表达量小于D、F组(P均<0.05)。E组培养液上清中HBsAg、HBeAg水平及细胞中HBV DNA表达低于B组,高于D组和F组(P均<0.05)。D组培养液上清中HBsAg、HBeAg水平及细胞中HBV DNA表达低于A组(P均<0.05),F组培养液上清中HBsAg、HBeAg水平及细胞中HBV DNA表达低于C组(P均<0.05)。结论 OMT对miR-122表达下调Hep G2.2.15细胞的抗HBV作用受到抑制,OMT可能通过调节miR-122的表达对HBV复制起抑制作用。Objective To observe the antiviral effect of oxymatrine（ OMT） on low expression of miR-122 and HBV infection of hepatocytes. Methods Hep G2. 2. 15 cells were cultured in vitro,and then were divided into groups A,B,C,D,E,and F. After 24 h,the culture medium was replaced with serum-free DMEM medium in the groups A,B and C,and that in the groups D,E,and F was replaced with serum-free medium containing 4 mg/m L OMT. At the same time,the cells in the groups A and D were added with Lipofectamine2000,cells in the groups B and E were added with Lipofectamine2000 and miR-122 inhibitor hsa-miR-122-5 p inhibitor,and cells in the groups C and F were added with Lipofectamine2000 and negative inhibitors. After 48 h,cells from each group were collected and miR-122 was detected by fluorescence quantitative PCR. The supernatant and cells were collected at 48 h after transfection. HBsAg and HBeAg in the supernatant were detected by electrochemiluminescence,and HBV DNA in the cells was detected by fluorescence quantitative PCR. Results The relative expression of miR-122 in the groups A,B,C,D,E and F were 1. 00 ± 0. 26,0. 72 ±0. 15,1. 04 ± 0. 28,3. 09 ± 0. 94,1. 27 ± 0. 59,and 3. 00 ± 0. 62,respectively. The relative expression of miR-122 in the group B was less than that in the groups A and C（ P〈0. 05）. The relative expression of miR-122 in the group E was less than that in the groups D and F（ P〈0. 05）. The levels of HBsAg and HBeAg in the supernatant and the expression of HBV DNA in cells of the group E were lower than those in the group B,which were higher than those in the groups D and F（ all P〈0. 05）. In the supernatant of group D,the level of HBsAg,HBeAg and the expression of HBV DNA in the cells were lower than those in the group A（ P〈0. 05）. The HBsAg,HBeAg level,and the HBV DNA expression in the culture medium of the group F were lower than those in the group C（ P〈0. 05）. Conclusions The anti-HBV effect of OMT is inhibited in Hep G2. 2. 15 cells with down-regulated expression of miR-1

关 键 词：氧化苦参碱 乙型肝炎病毒 微小RNA-122 

分 类 号：R575.1[医药卫生—消化系统]