摘要
目的:探讨自噬在可溶性尿酸(UA)诱导肾小管上皮细胞(HK-2细胞)炎症反应中的作用。方法:(1)分别用可溶性UA、自噬增强剂雷帕霉素(RAP)联合可溶性UA和自噬抑制剂氯喹(CQ)联合可溶性UA刺激HK-2细胞,透射电镜观察自噬体,western blotting法检测单核细胞趋化蛋白-1(MCP-1)、自噬微管相关的轻链蛋白3(LC3-Ⅱ)和自噬相关调控基因蛋白Beclin1的表达;(2)用Akt抑制剂Perifosine与可溶性UA溶液共刺激HK-2细胞,western blotting法检测p-Akt和MCP-1的表达。结果:与对照组比较,可溶性UA可诱导HK-2细胞MCP-1、LC3-Ⅱ和Beclin1表达上调(均P<0.05),透射电镜观察到自噬体数量增加。与UA组比较,RAP与可溶性UA共刺激HK-2细胞后,自噬增强而MCP-1表达减弱(均P<0.05),CQ与可溶性UA作用HK-2细胞后,自噬抑制而MCP-1表达增加(均P<0.05)。与单纯UA刺激的HK-2细胞比较,p-Akt阻断剂Perifosine联合UA共刺激细胞后MCP-1表达减弱(P<0.05)。结论:自噬参与调控可溶性UA诱导的肾小管上皮细胞炎症反应。
Objective：To investigate the role of autophagy in soluble uric acid（UA）-induced inflammatory response in renal tubular epithelial cells（HK-2 cells）.Methods：Cells were pretreated with or without UA in soluble form and then stimulated with rapamycin（RAP,an autophagy enhancer）or chloroquine（CQ,an autophagy inhibitor）.The changes of autophagosomes were observed under transmission electron microscopy.The expressions of monocyte chemoattractant protein-1（MCP-1）,autophagic microtubule-associated light chain protein 3（LC3-Ⅱ）and autophagy-related protein Beclin1 were determined by western blotting.Subsequently,HK-2 cells were co-stimulated with soluble UA and an Akt inhibitor perifosine.The expressions of phosphor-Akt（p-Akt）and MCP-1 were detected by western blotting as well.Results：Soluble UA treatment resulted in up-regulation of MCP-1,LC3-Ⅱ,and Beclin1 in HK-2 cells（P〈0.05）and an increase in the number of autophagosomes.The LC3-Ⅱand Beclin1 expressions were enhanced while the MCP-1 expression was down-regulated in HK-2 cells after stimulation with RAP（P〈0.05）,but these changes were reversed by CQ stimulation（P〈0.05）.The expression level of MCP-1 was decreased after stimulation of pAkt inhibitor perifosine compared with non-treated control（P〈0.05）.Conclusion：Autophagy might be involved in the soluble UA-induced inflammation in renal tubular epithelial cells.
作者
陈园园
易扬
覃廖缓
刘福岗
谢恺庆
杨海波
Chen Yuanyuan;Yi Yang;Qin Liaoyuan;Liu Fugang;Xie Kaiqing;Yang Haibo(Department of Nephrology and Blood Purification,the Second Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Department of Microbiology,Guangxi Medical University,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2018年第6期756-761,共6页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No.81360243)
广西自然科学基金资助项目(No.2013GXNSFAA019149)
关键词
自噬
可溶性尿酸
肾小管上皮细胞
炎症
autophagy
soluble uric acid
tubular epithelial cells
inflamination
