丙酮醛对人近曲小管上皮细胞氧化应激状态及NLRP3炎症小体信号的作用  

Effects of methylglyoxal on oxidative stress and NLRP3 inflammasome signaling in HK-2 cells

作　　者：陈燕玲[1] 罗婷 曾洪敏 吴秀香[1] 凡栋 庄晓东[2] 黄霜枝 王晓玲[1] 

机构地区：[1]遵义医学院珠海校区基础教学部,珠海理学博士519041 [2]中山大学附属第一医院心血管内科,广州510080

出　　处：《医学研究生学报》2018年第3期262-266,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81560133;81600206);贵州省科技合作计划项目(黔科合LH字[2015]7529号);广东省自然科学基金(2016A030310140);遵义医学院博士启动基金(F-711)

摘　　要：目的丙酮醛引起人近曲小管上皮细胞(HK-2细胞)的损伤机制尚未明确。文中旨在观察丙酮醛对HK-2细胞氧化应激状态及NLRP炎症小体信号的影响,探讨其引起HK-2细胞损伤的机制。方法取对数生长期的HK-2细胞分为5组:对照组(采用含10%胎牛血清的DMEM/F12培养基培养),100、200、400、800、1600μmol/L丙酮醛组(分别加入100、200、400、800、1600μmol/L的丙酮醛处理24 h),应用硫代巴比妥酸法检测超氧化物歧化酶(SOD)水平,试剂盒法测定乳酸脱氢酶(LDH)的活力,DCFH-DA染色法测定细胞内活性氧(ROS)水平,Western blot法检测HK-2细胞内NLRP3、caspase-1、IL-1β及NF-κB蛋白表达水平。结果与对照组SOD活力[(975.12±12.82)U/m L]比较,100、200、400、800、1600μmol/L丙酮醛组[(797.48±33.08)、(720.00±24.35)、(384.57±17.53)、(130.39±9.16)、(77.80±11.78)U/m L]明显降低(P<0.05);与对照组LDH水平[(808.82±31.51)U/L]比较,100、200、400、800、1600μmol/L丙酮醛组[(908.87±11.01)、(976.21±17.37)、(1161.15±38.44)、(1451.16±24.62)、(1832.98±36.52)U/L]显著升高(P<0.05)。与对照组比较,100、200、400、800、1600μmol/L丙酮醛组炎症小体相关组分NLRP3、caspase-1蛋白以及caspase-1下游蛋白IL-1β、NF-κB蛋白表达水平明显升高(P<0.05)。结论丙酮醛可通过氧化应激及激活NLRP3炎症小体信号引起HK-2细胞损伤。Objective Methylglyoxal can cause the injury of human proximal tubular epithelial cell line（HK-2 cells）,but the exact mechanism is still unclear.The present study aimed to explore the influence of oxidative stress and the expression of NLRP3 inflammasome in HK-2 cells induced by methylglyoxal.Methods HK-2 cells at logarithmic phase were divided into six groups：control group and 100,200,400,800,1600 μmol/L methylglyoxal groups（cells were cultured in 100,200,400,800,1600 μmol/L methylglyoxal concentration for 24 h）.Superoxide dismutase（SOD） levels were assayed by thibabituric acid method.Release of lactate dehydrogenase（LDH） activity was detected by assay kit.ROS production was measured by DCFH-DA staining.The expression levels of NLRP3,caspase-1,IL-1β and NF-κB were evaluated by western blot.Results Compared with control group,different methylglyoxal concentrations could enhance ROS level and LDH activity in HK-2 cells（P0.05） and reduce SOD level significantly（P0.05）.The results of western blot showed the protein levels of NLRP3,caspase-1,IL-1β and NF-κB were significant up-regulated after the addition of methylglyoxal（P0.05）.Conclusion Methylglyoxal may induce the injury of HK-2 cells by oxidant stress and activating of NLRP3 inflammasome signaling.

关 键 词：丙酮醛 氧化应激 NLRP3炎症小体 HK-2细胞 

分 类 号：R691.9[医药卫生—泌尿科学]