论文部分内容阅读
目的探讨p53凋亡刺激蛋白2(apoptosis-stimulating protein of p53-2,ASPP2)对人肝星状细胞活化的调节作用及机制。方法人永生的肝星状细胞系LX-2在孵箱中培养,转染ASPP2腺病毒和单荧光自噬指示体系(GFP-LC3)质粒,免疫印迹法检测ASPP2、α-平滑肌肌动蛋白(α-SMA)、自噬基因表达相关蛋白(Beclin-1)的表达水平。免疫荧光检测,LX-2细胞共转染ASPP2腺病毒和GFP-LC3质粒后,ASPP2过表达对LX-2细胞自噬的影响,M30染色检测ASPP2过表达对LX-2细胞凋亡的影响。结果转染ASPP2腺病毒后LX-2细胞内ASPP2蛋白表达明显增加;与转染对照空载腺病毒的LX-2细胞相比,转染ASPP2腺病毒的LX-2细胞自噬和α-SMA的表达明显减少,肝星状细胞的活化受到抑制;转染ASPP2腺病毒后LX-2细胞的凋亡水平增加明显。结论 ASPP2过表达在体外具有抑制肝星状细胞活化的作用。
Objective To investigate the regulatory effect of ASPP2 on the activation of human hepatic stellate cells and its mechanism. Methods Human immortalized hepatic stellate cell line LX-2 was cultured in an incubator and transfected with the ASPP2 adenovirus and the single fluorescent autophagy indicator system (GFP-LC3). Western blotting was used to detect the expression of ASPP2, α-smooth muscle actin α-SMA) and the expression of autophagy gene expression-related protein (Beclin-1). The effect of ASPP2 overexpression on LX-2 cell autophagy was detected by immunofluorescence assay. The effect of ASPP2 overexpression on LX-2 cell apoptosis was detected by M30 staining. Results The expression of ASPP2 protein in LX-2 cells was significantly increased after transfection with ASPP2 adenovirus. Compared with LX-2 cells transfected with control adenovirus, Significantly decreased the activation of hepatic stellate cells was inhibited; transfected with ASPP2 adenovirus LX-2 cells increased significantly. Conclusion ASPP2 overexpression can inhibit the activation of hepatic stellate cells in vitro.