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

发布时间：2018-05-15 01:25

本文选题：腺苷A受体 + 腺苷AA受体　；参考：《中国药理学通报》2015年01期

【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。
[Abstract]:Aim to investigate the effects of adenosine A1 receptor (A1R) and adenosine A2A1R (A2A2ARR) on HSC activation and proliferation induced by acetaldehyde in rat hepatic stellate cells (HSCs). Methods A rat model of alcoholic liver fibrosis (HSC) was established by acetaldehyde induced HSC-T6 in vitro. A1R and A2AR small interfering RNA(small interfering siRNAs were designed and synthesized. The transfection efficiency of A1R and A2AR small interfering RNA(small interfering siRNAs was observed by fluorescence inverted microscope, and transient transfection of A1R and A2AR into HSC-T6 cells was observed by liposome LipofectamineTM2000. The proliferation of HSC-T6 cells was detected by MTT assay, and the expression of A1RN A2ARand 伪 -SMACollagen I mRNA and protein of HSC-T6 were detected by Real-Time Q PCR and Western blot, respectively. Results the expression levels of A1R and A2AR genes and proteins in HSC-T6 cells were significantly decreased after transfection of A1R and A2AR siRNA, and the expression levels of 伪 -SMA-Collagen I mRNA and protein were also significantly decreased. Blocking the expression of A1R or A2AR genes could significantly inhibit the activation and proliferation of HSC-T6 cells. Conclusion targeting blocking the expression of A1R or A2AR gene can significantly inhibit the activation and proliferation of HSC-T6 cells. A1R and A2AR may be potential therapeutic targets for alcoholic liver fibrosis.
【作者单位】：安徽医科大学药学院安徽医科大学肝病研究所;
【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)
【分类号】：R575.2

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