·糖尿病基础研究·

　　【摘要】 目的 观察高糖诱导大鼠肾小管上皮细胞(NRK-52E细胞)NLRP3炎性小体及炎症因子蛋白的表达，并通过转染NLRP3特异性小干扰质粒(siRNA)明确NLRP3炎性小体是否参与其炎症损伤。 方法 (1)将NRK-52E细胞分为对照(Con)组、高糖(HG)6 h、12 h、24 h、48 h组。用Western Blot检测NLRP3、凋亡相关斑点样蛋白(ASC)及天冬氨酸蛋白水解酶1(caspase-1)的表达。(2)根据上述结果分为Con组、HG组、HG+NLRP3-siRNA质粒组、HG+无关质粒组，培养48 h后RT-PCR检测 NLRP3 mRNA的表达;Western Blot检测NLRP3、ASC、白介素1β(IL-1β)、白介素6(IL-6)、肿瘤坏死因子-a(TNF-a)蛋白表达。 结果 (1)HG刺激后的NRK-52E细胞NLRP3、ASC蛋白表达逐渐增加，48 h组最为明显(P均<0.05);caspase-1蛋白表达较Con组明显增高(P<0.05)。(2)HG组NLRP3、ASC、IL-1β、IL-6、TNF-a蛋白表达较Con组明显增高(P<0.05);HG+NLRP3-siRNA质粒组上述蛋白表达较HG组均显著下降(P<0.05)。 结论 HG可诱导NRK-52E细胞NLRP3炎性小体活化，进而促进其下游炎症因子的表达和释放。推测NLRP3炎性小体可能参与了HG诱导的NRK-52E细胞的炎症损伤。

　　【关键词】 NLRP3炎性小体;高糖;肾小管上皮细胞;炎性反应

The changes of NLRP3 inflammasome in the high glucose-induced inflammatory response of renal tubular epithelial cells ZHAO Hong-yu, LI Rong-shan, YU Wei-min, et al. Department of Nephrology, the Second Affiliated Hospital, Shanxi Medical University, Taiyuan 030001, China

Corresponding author: LI Rong-shan, E-mail: rongshanli@yahoo.com.cn

　　【Abstract】Objective To investigate the expressions of NLRP3 inflammasome and inflammatory factor protein in the high glucose-induced renal tubular epithelial cells (NRK-52E cells) and to make clear its effect on inflammatory damage through transfection of small interfering RNA (siRNA). Methods (1). The NRK-52E cells were divided into control group and high glucose (HG) 6h, 12h, 24h, and 48h groups. Western blotting was adopted to detect the expressions of NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), and Caspase-1. (2). NRK-52E cells were divided into the control group, high glucose group, HG+NLRP3-siRNA group, and HG+irrespective siRNA group on the basis of the result of above-mentioned tests. After 48h fostering, the expression of NLRP3 mRNA was detected by RT-PCR, and the expressions of NLRP3, ASC, IL-1β, IL-6, and TNF-a proteins were determined by Western blotting. Results (1). The expressions of NLRP3 and ASC were gradually increased, and the highest expression was in the 48h group after stimulated by high glucose (P<0.05); the expression of Caspase-1 was significantly higher than that in the control group (P<0.05). (2). Compared with the control group, the expressions of NLRP3, ASC, IL-1β，IL-6, and TNF-a proteins were significantly increased in the HG group (P<0.05); these expressions in the HG+NLRP3-siRNA group were significantly decreased than those in the HG groups (P<0.05). Conclusion High glucose induces the activation of NLRP3 inflammasome in the NRK-52E cells, leading to the expression and release of its downstream inflammatory factors. It is assumed that the NLRP3 inflammasome may have been involved in the high glucose-induced inflammatory damage in NRK-52E cells.

　　【Key words】NLRP3 inflammasome; High glucose; Renal tubular epithelial cells; Inflammatory reaction