C3a-C3aR在尿酸促进人肾小管上皮细胞MCP-1表达中的作用  

Uric Acid Induces MCP-1 in Renal Tubular Epithelial Cells through C3a-C3aR Interaction

作　　者：尤丹瑜[1] 罗丛伟 万建新[1] 

机构地区：[1]福建医科大学附属第一医院肾内科,福州350005

出　　处：《福建医科大学学报》2015年第6期355-359,共5页Journal of Fujian Medical University

基　　金：福建省卫生厅青年科研课题(2012-1-25)

摘　　要：目的探讨尿酸(UA)对人肾小管上皮细胞单核趋化蛋白-1(MCP-1)表达的影响及其机制。方法体外培养人肾皮质近曲小管上皮细胞(HK-2),以不同浓度UA分别作用不同时间,利用逆转录聚合酶链反应(RT-PCR)检测MCP-1的mRNA水平。在UA诱导下,通过针对补体C3的小干扰RNA(C3siRNA)转染下调HK-2细胞内源性C3的表达、小分子C3aR阻断剂SB290157阻断C3a-C3aR作用、外源性C3a刺激促进C3aR激活3种方式抑制或激活C3aR的信号通路,并通过实时荧光定量PCR(Q-PCR)检测其对于MCP-1mRNA表达的影响。结果 150μmol/L UA干预HK-2细胞12h可上调MCP-1mRNA转录水平;C3siRNA转染或C3aR阻断能够抑制被UA上调的MCP-1mRNA的表达;C3a与UA共同进行干预时,显著增强UA对MCP-1mRNA的诱导作用。结论 UA可上调HK-2细胞MCP-1mRNA的表达,其机制可能与UA激活C3或C3aR有关。Objective To investigate the role of the interaction of complement fragment 3a（C3a）and its receptor C3 aR in uric acid（UA）-induced expression of monocyte chemoattractant protein-1（MCP-1）in human renal proximal tubular epithelial cell line HK-2. Methods HK-2cells were cultured in vitro,stimulated by UA of different concentrations for different lengths of times. Reverse transcription polymerase chain reaction（RT-PCR）was performed to analyze the transcription of MCP-1mRNA.Three methods were used to change the C3 aand its receptor＇s interaction after the stimulation of UA,including silencing endogenous C3 expression with small interfering RNA（siRNA）targeted C3,blocking C3 aR with small molecule antagonist SB290157,and activating the C3 aR with synthetic C3 a. Real-time quantitative PCR was used to analyze the expression of MCP-1 mRNA in these conditions. Results150μmol/L UA stimulating for 12 hsignificantly up-regulated HK-2cells＇ MCP-1mRNA transcription. The over-expression of MCP-1induced by UA could be inhibited by C3 siRNA transfection or C3 aR blockage. When C3 astimulated HK-2cells,together with UA,it could further elevate UA-induced MCP-1mRNA expression. Conclusion The activation of HK-2cells＇ C3 aR provides the important stimulating signal for MCP-1expression. Blocking the interaction of C3a-C3 aR will significantly inhibit UAinduced MCP-1expression.

关 键 词：肾小管上皮细胞 单核趋化蛋白-1 尿酸 

分 类 号：R692[医药卫生—泌尿科学]