论文部分内容阅读
目的研究NLRP3炎症复合体在高糖培养的大鼠近端肾小管上皮细胞(NRK-52E)中的表达及其与细胞凋亡的关系。方法将NRK-52E细胞分为正常组、等高对照组、高糖组、高糖+阴性对照组及高糖+干扰组。培养48h后应用Western blot检测各组NLRP3,凋亡相关斑点样蛋白(ASC)及天冬氨酸蛋白水解酶1、3(Caspase-1、3)的表达情况。细胞凋亡情况采用流式细胞仪检测。结果高糖刺激可上调NRK-52E细胞NLRP3炎症复合体的表达及Caspase-3的表达(P均<0.05),细胞凋亡率升高(P<0.05);通过特异性的SiRNA抑制NLRP3蛋白质的表达后,细胞NLRP3炎症复合体及Caspase-3的表达均减少,细胞凋亡率下降(P均<0.05)。结论高糖促进NRK-52E细胞NLRP3炎症复合体表达,针对NLRP3的SiRNA通过减少NLRP3炎症复合体的表达从而抑制高糖诱导的细胞凋亡。
Objective To investigate the expression of NLRP3 inflammasome in human proximal tubular epithelial cells (NRK-52E) cultured in high glucose and its relationship with apoptosis. Methods NRK-52E cells were divided into normal group, control group, high glucose group, high glucose + negative control group and high glucose + interference group. Western blot was used to detect the expression of NLRP3, apoptotic related spot-like protein (ASC) and aspartate protease 1, 3 (Caspase-1, 3) after 48 h culture. Apoptosis was detected by flow cytometry. Results High glucose stimulated the expression of NLRP3 inflammatory complex and the expression of Caspase-3 in NRK-52E cells (all P <0.05), and increased the apoptosis rate (P <0.05). The inhibition of NLRP3 protein by specific siRNA After the expression, the expression of NLRP3 inflammatory complex and Caspase-3 decreased and the apoptosis rate decreased (all P <0.05). Conclusion High glucose can promote the expression of NLRP3 inflammatory complex in NRK-52E cells. SiRNA targeting NLRP3 can inhibit high glucose-induced apoptosis by decreasing the expression of NLRP3 inflammatory complex.