论文部分内容阅读
目的观察高糖诱导大鼠肾小管上皮细胞(NRK-52E)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(NC)组及高糖(HG)6、12、24、48h组。用Western Blot检测NL-RP3、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NC组、HG组、HG+NLRP3-siRNA质粒组、HG+无关质粒组,培养48h后RT-PCR检测NLRP3mR-NA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-α(TNF-α)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,48h组最为明显(P均<0.05);caspase-1蛋白表达较NC组增高(P<0.05)。(2)HG组NLRP3、ASC、IL-1β、IL-6、TNF-α蛋白表达较NC组增高(P<0.05);HG+NLRP3-siRNA质粒组上述蛋白表达较HG组均下降(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。
Objective To investigate the expression of NLRP3 inflammasome and inflammatory cytokines in high glucose induced rat renal tubular epithelial cells (NRK-52E) and to determine if NLRP3 inflammasome is involved in the expression of NLRP3 specific small interfering plasmid (siRNA) Inflammatory damage. Methods (1) NRK-52E cells were divided into control (NC) group and high glucose (HG) 6,12,24,48h groups. Western Blot was used to detect the expression of NL-RP3, apoptosis related ASC and caspase-1. (2) The expression of NLRP3mR-NA was detected by RT-PCR after cultured for 48h. The expression of NLRP3, ASC, interleukin 1β (IL-6) IL-1β), interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) protein. Results (1) The expression of NLRP3 and ASC protein in NRK-52E cells increased gradually after HG stimulation, which was the most obvious in 48h group (all P <0.05). The protein expression of caspase-1 in NRK-52E cells was increased compared with NC group (P <0.05). (2) The expression of NLRP3, ASC, IL-1β, IL-6 and TNF-αin HG group was higher than that in NC group (P <0.05) 0.05). Conclusion HG can induce the activation of NLRP3 inflammasome in NRK-52E cells and further promote the expression and release of inflammatory factors downstream of it. It is speculated that NLRP3 inflammasome may be involved in HG-induced inflammatory damage of NRK-52E cells.