RNAi介导大鼠腺苷A1和A2A受体基因沉默对乙醛诱导的肝星状细胞活化增殖的影响  

Proliferation and activation of acetaldehyde-induced HSC-T6 cells through RNA inference targeting adenosine A1 and A2A receptors

作　　者：王琪[1] 王和[1] 饶玲[1] 赵晗[1] 杨凤[1] 杨岩[1] 吕雄文[1] 李俊[1] 

机构地区：[1]安徽医科大学药学院,安徽医科大学肝病研究所,安徽合肥230032

出　　处：《中国药理学通报》2015年第1期50-55,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81270498);安徽省自然科学基金项目(No 11040606M194);安徽高校省级科学研究重点项目(No KJ2012A148);国家级大学生创新创业训练计划项目(No 201310366032)

摘　　要：目的研究大鼠腺苷A1受体(A1R)和腺苷A2A(A2AR)受体的siRNA分别转染大鼠肝星状细胞(HSC)对乙醛诱导的HSC活化增殖的影响。方法采用乙醛诱导HSC-T6建立离体的大鼠酒精性肝纤维化HSC模型,设计并合成A1R和A2AR小干扰RNA(small interfering RNA,siRNA)序列,通过脂质体LipofectamineTM2000瞬时转染至HSC-T6细胞内,荧光倒置显微镜观察细胞的转染效率,用四甲基偶氮唑盐(MTT)法检测HSC-T6细胞增殖变化;利用Real-Time q PCR及Western blot法分别检测HSC-T6的A1R、A2AR、α-SMA、Collagen I mRNA及蛋白表达。结果将A1R和A2AR siRNA转染至HSC-T6细胞内,A1R和A2AR基因及蛋白的表达水平明显降低;同时α-SMA、Collagen I mRNA及蛋白表达水平亦明显降低;靶向封闭A1R或A2AR基因的表达可明显抑制HSC-T6细胞的活化增殖。结论靶向封闭A1R或A2AR基因的表达可明显抑制HSC-T6细胞的活化增殖,A1R和A2AR可能是潜在的酒精性肝纤维化的治疗靶点。Aim To investigate the influence of down-regulating adenosine A1 receptor and adenosine A2 A receptor gene expression on proliferation and activation of acetaldehyde-induced hepatic stellate cell-T6 cells through siRNA. Methods Alcoholic liver fibrosis in vitro model was constructed by inducing HSC-T6 cells with acetaldehyde. siRNA targeting A1R and A2AR were designed and synthesized according to its mRNA. The siRNA was transfected into rat HSC-T6 cells by li-posome LipofectamineTM 2000. HSC cell proliferation was measured by MTT. The mRNA levels of A1R, A2AR, α-SMA, Collagen I in the supernatant of the cell culture were measured by Quantitative Real-Time PCR. The protein levels of A1R, A2AR, α-SMA,Collagen I were measured by Western blot. Results A1 R and A2 AR siRNA effectively inhibited the cell proliferation, and they also significantly decreased the levels of A1R, A2AR,α-SMA, Collagen I, suggesting that A1 R and A2 AR might be potential target genes in the alcoholic liver fibrosis. Conclusions Silencing A1 R or A2 AR by RNAi can significantly inhibit the HSC proliferation, A1R and A2AR may be potential therapeutic target genes for alcoholic liver fibrosis.

关 键 词：腺苷A1受体 腺苷A2 A受体 肝星状细胞 细胞增殖 乙醛 酒精性肝纤维化 

分 类 号：R-332[医药卫生] R322.47