白藜芦醇抑制TXNIP/NLRP3信号通路改善脂多糖诱导的肾小管上皮细胞炎性损伤

Effects of resveratrol against inflammatory injury of human renal proximal tubular cells induced by LPS through the inhibition of TXNIP/NLRP3 signaling pathway

关键词:  白藜芦醇  肾小管上皮细胞  炎性反应  TXNIP/NLRP3信号通路

Key words:  resveratrol  human renal proximal tubular cells  inflammatory response  TXNIP/NLRP3 signaling pathway

摘要:

[目的]观察白藜芦醇对脂多糖诱导的肾小管上皮细胞（HK-2）炎性损伤的保护作用，及其对TXNIP/NLRP3炎性通路的影响。[方法]采用脂多糖（LPS，1 mg/L）体外诱导HK-2细胞炎性损伤模型，将细胞分为正常对照组、LPS诱导组、LPS+低剂量（50 μmol/L）白藜芦醇组、LPS+中剂量（100 μmol/L）白藜芦醇组、LPS+高剂量（200 μmol/L）白藜芦醇组；CCK8法检测HK-2细胞相对存活率，酶联免疫吸附测定（ELISA）法检测炎性因子白介素-6（IL-6）、白介素-1β（IL-1β）及肿瘤坏死因子-α（TNF-α）水平，Western Blot方法分析诱导型一氧化氮合酶（iNOS）、环氧酶-2（COX-2）、人硫氧还蛋白互作蛋白（TXNIP）及人隐热蛋白（NLRP3）表达。[结果]与LPS诱导组比较，不同浓度白藜芦醇干预的HK-2细胞相对存活率明显升高（P<0.05），细胞上清炎性因子IL-6、IL-1β及TNF-α水平明显降低（P<0.05），细胞中炎性蛋白iNOS及COX-2表达水平显著降低（P<0.05）；细胞中TXNIP及NLRP3蛋白表达也显著降低（P<0.05），并表现出剂量依赖性。[结论]白藜芦醇可以明显抑制LPS诱导的HK-2细胞炎症损伤，抑制炎症相关蛋白的表达，其作用机制可能与抑制TXNIP/NLRP3炎性通路活化相关。

Abstract:

[Objective] To observe the protective effects of resveratrol against inflammatory damage of HK-2 cells induced by LPS,and observe the effect of resveratrol on the activation of TXNIP/NLRP3 signaling pathway.[Methods] LPS (1 mg/L) was used to induce the model of inflammatory injury in HK-2 cells in vitro. HK-2 cells were divided into five groups,including control group,LPS-induced group,LPS+low dose (50 μmol/L) resveratrol group,LPS+middle dose (100 μmol/L) resveratrol group,and LPS+high dose (200 μmol/L) resveratrol group. CCK8 assay was used to measure the relative survival rates of HK-2 cells. ELISA assay was used to detect the levels of IL-6,IL-1β and TNF-α. Western Blot was used to determine the expression levels of iNOS,COX-2,TXNIP and NLRP3 proteins.[Results] Compared with LPS induced group,the relative survival rates of HK-2 cells were greatly increased after treatment with different concentrations of resveratrol (P<0.05). The levels of inflammatory factors (IL-6,IL-1β and TNF-α) were significantly decreased (P<0.05). The expression levels of iNOS,COX-2,TXNIP and NLRP3 were also greatly decreased (P<0.05) with a dose-dependent manner.[Conclusion] Resveratrol can significantly inhibit the inflammatory response of HK-2 cells induced by LPS via suppressing the activation of TXNIP/NLRP3 signaling pathway.