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白藜芦醇通过TXNIP-NLRP3通路对HK-2细胞高糖缺氧复氧损伤的作用
作者:肖业达1  曹红2  赵博1  黄亚医1  汪华新1 
单位:1. 武汉大学人民医院 麻醉科, 湖北 武汉 430060;
2. 武汉市第三医院 麻醉科, 湖北 武汉 430060
关键词:硫氧还蛋白相互作用蛋白 炎性体3 高糖 缺氧复氧 白藜芦醇 人肾小管上皮细胞 
分类号:R692.5
出版年·卷·期(页码):2018·46·第一期(15-18)
摘要:

目的:探讨白藜芦醇通过硫氧还蛋白相互作用蛋白(TXNIP)-炎性体3(NLRP3)通路对人肾小管上皮细胞(HK-2细胞)高糖缺氧复氧损伤的作用。方法:随机将HK-2细胞分为高糖组(HG组)、高糖缺氧复氧组(HHR组)及高糖缺氧复氧+白藜芦醇预处理组(HHR-RES组)3组,每组n=5。采用高糖刺激72 h,缺氧4 h复氧2 h的方法建立高糖缺氧复氧模型,白藜芦醇预处理组在高糖刺激的同时给予白藜芦醇50 μmol·L-1处理。CCK-8和乳酸脱氢酶(LDH)法分别检测细胞存活率和细胞损伤情况,超氧化物歧化酶(SOD)和丙二醛(MDA)法检测细胞氧化应激水平,流式细胞术检测细胞凋亡率,免疫荧光法检测TXNIP和NLRP3蛋白的表达。结果:与HG组比较,HHR组LDH水平、MDA含量、细胞凋亡率、TXNIP和NLRP3蛋白表达升高(均P<0.05),而细胞存活率、SOD活性降低(均P<0.05);与HHR组比较,HHR-RES组LDH水平、MDA含量、细胞凋亡率、TXNIP和NLRP3蛋白表达降低(均P<0.05),而细胞存活率、SOD活性升高(均P<0.05)。结论:白藜芦醇通过抑制TXNIP-NLRP3通路减轻HK-2细胞在高糖缺氧复氧下的损伤。

Objective:To evaluate the role of resveratrol on human renal proximal tubular cells HK-2 injury induced by high glucose and hypoxia/reoxygenation and the its possible relationship with TXNIP-NLRP3 pathway.Methods:HK-2 cell were randomly divided into 3 groups:the high glucose group (group HG),the high glucose and hypoxia/reoxygenation group(group HHR) and the high glucose and hypoxia/reoxygenation+resveratrol group(group HHR-RES),each n=5.The high glucose and hypoxia/reoxygenation model was established by stimulated with high glucose for 72 h,then exposed to hypoxia 4 h and reoxygenation 2 h. In the HHR-RES group, 50 μmol·L-1 of resveratrol was treated during the 72 h of high glucose incubation. The cell viability and cell damage was detected by CCK-8 and LDH respectively, oxidative stress was detected by SOD and MDA, the apoptotic rate of cells was detected by flow cytometry,the immunofluorescence staining was performed to detect the expression of TXNIP and NLRP3. Results:Compared with the group HG, the levels of LDH and MDA,apoptotic rate,the expression of TXNIP and NLRP3 were increased(all P<0.05);cell viability and SOD activity were decreased in the group HHR (all P<0.05),Compared with the group HHR,the levels of LDH and MDA, apoptotic rate,the expression of TXNIP and NLRP3 were decreased(all P<0.05);cell viability and SOD activity were increased in the group HHR+RES (all P<0.05). Conclusion:Resveratrol may inhibit TXNIP-NLRP3 pathway to alleviate the HK-2 cell injury induced by high glucose and hypoxia/reoxygenation.

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