晚期糖基化终产物诱导人肾小管上皮细胞TAK1活化的机制  

The mechanism of TAK1 activation induced by advanced glycation end products in human renal tubular epithelial cells

作　　者：卢守燕[1] 贾京京 黄文辉[1] 董丽婷 李莹屏[1] 马志刚[1] LU Shouyan;JIA Jingjing;HUANG Wenhui(Department of Nephrology,Gansu Provincial Hospital,Lanzhou 730000,CHINA)

机构地区：[1]甘肃省人民医院肾内科,甘肃兰州730000 [2]康县第一人民医院肾内科

出　　处：《江苏医药》2022年第4期333-336,共4页Jiangsu Medical Journal

基　　金：甘肃省人民医院院内科研基金项目(18GSSY5-22)。

摘　　要：目的探讨晚期糖基化终产物(AGEs)诱导人近端肾小管上皮细胞系HK-2中转化生长因子β激活激酶1(TAK1)活化的作用机制.方法将HK-2细胞于含10%胎牛血清的DMEM/F12培养基中培养;待细胞增殖达到70%~80%时,换用无血清培养基稳定静止24 h后分组处理.分别用不同浓度AGE-牛血清白蛋白(AGE-BSA)培养24、48、72 h,共分为四组:A组(AGE-BSA 0μg/mL处理),B1组(AGE-BSA 20μg/mL处理),B2组(AGE-BSA 50μg/mL处理),B3组(AGE-BSA 100μg/mL处理).分别采用ELISA法、RT-PCR法和Western blot法检测AGE-BSA刺激HK-2细胞分泌TAK 1含量、mRNA和蛋白的表达.结果随着AGE-BSA处理浓度(20、50和100μg/mL)的升高,TAK1在HK-2细胞中含量、mRNA和蛋白表达逐渐增加;尤其B3组刺激72 h时TAK1含量、mRNA和蛋白表达最高(P<0.05).结论AGEs环境可诱导HK-2细胞增殖,激活细胞内TAK1信号传导通路,对细胞增殖过程中TAK1蛋白及mRNA的表达有上调作用.Objective To investigate the activation mechanism of TGF-βactivated kinase 1(TAK1)in human renal tubular epithelial cell line HK-2 cultured with advanced glycation end products(AGEs).Methods HK-2 cells were cultured in vitro in medium(DMEM/F12)containing 10%fetal bovine serum.When the cell proliferation reached to 70%-80%,the serum-free medium was changed to a stable static medium for 24 hours.And then the cells were divided into four groups of A(treated with AGE BSA 0 μg/mL),B1(treated with AGE-BSA 20μg/mL),B2(treated with AGE-BSA 50 μg/mL),B3(treated with AGE-BSA 100 μg/mL)and cultured for 24,48 and 72 h,respectively.The content,mRNA and protein expressions of TAK1 secreted by HK-2 cells stimulated by AGE-BSA were detected by ELISA,RT-PCR and Western blot,respectively.Results With the increase of AGE-BSA concentrations(20,50,and 100 μg/mL),the content,mRNA and protein expressions of TAK1 in HK-2 cells were gradually increased.The increase of the content,mRNA and protein expressions TAK1 were most significant in group B3(P<0.05).Conclusion AGEs can induce the activation and proliferation of HK-2 cells,activate the intracellular TAK1 signaling pathway,and up-regulate the protein and mRNA expressions of TAK1 during cell proliferation.

关 键 词：肾小管上皮细胞 细胞增殖 转化生长因子β激活激酶1 

分 类 号：R692[医药卫生—泌尿科学]