TAK1抑制剂对高糖环境巨噬细胞和系膜细胞相互作用的影响及机制  

Effect of TGF-β activated kinase-1 inhibitor on the interaction between macrophages and mesangial cells on the condition of high glucose

作　　者：卫洁[1] 冯世尧 徐兴欣[1] 吴永贵[1] Wei Jie Feng Shirao Xu Xingxin et al(Dept of Nephropathy, The First Affiliated Hospital of Anhui Medical University,Hefei 23002)

机构地区：[1]安徽医科大学第一附属医院肾内科,合肥230022

出　　处：《安徽医科大学学报》2016年第10期1453-1458,1463,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81270813)

摘　　要：目的探讨高糖环境下转化生长因子β激活激酶1(TAK1)抑制剂5Z-7-oxozeaenol在巨噬细胞和系膜细胞相互关系中的作用。方法巨噬细胞和系膜细胞共培养、巨噬细胞单培养、系膜细胞单培养3种培养方式分别分为:抑制剂对照组、甘露醇对照组、正常对照组、高糖组、高糖+不同浓度(30、100、300 nmol/L)抑制剂组。采用Western blot法检测单培养巨噬细胞p-TAK1、TAK1结合蛋白(TAB1)、NF-κB p65蛋白表达变化,免疫荧光检测TAK1抑制剂对NF-κB p65亚基核转位的影响。采用ELISA法检测单培养和共培养各组细胞上清液中肿瘤坏死因子(TNF)-α、白介素(IL)-1β、单核细胞趋化因子(MCP)-1、Ⅳ型胶原(ColⅣ)、纤维连接蛋白(FN)含量变化,光镜下观察共培养对巨噬细胞迁移能力影响。结果与对照组比较,高糖刺激巨噬细胞pTAK1、TAB1、NF-κB p65蛋白表达均明显上调(P<0.05),高糖组共培养和单培养细胞上清液中TNF-α、IL-1β、MCP-1、ColⅣ、FN均显著高于对照组(P<0.05),共培养细胞上清液中各种细胞因子含量较单培养升高更明显(P<0.05),与高糖组比较,TAK1抑制剂呈浓度依赖性降低各项指标表达(P<0.05)。300 nmol/L抑制剂明显降低巨噬细胞迁移数目(P<0.05)。结论高糖环境下,巨噬细胞与系膜细胞之间存在相互影响,这种作用能促进炎症因子(TNF-α、IL-1β、MCP-1)和细胞外基质成分(ColⅣ、FN)产生增加,而TAK1抑制剂能通过干预NF-κB p65核转位,抑制巨噬细胞迁移,减少炎症因子和细胞外基质成分的分泌,发挥抗炎、抗纤维化效应,从而起到肾脏保护作用。Objective To investigate the effect of TGF-β activated kinase-1 inhibitor 5Z-7-oxozeaenol on the inter- action between macrophages and mesangial cells on the condition of high glucose. Methods The macrophages and mesangial cells were cultured separately or co-cultured and divided into seven groups： inhibitor control group, man- nitol control group, normal control group, high glucose group and high glucose ＋ inhibitor groups （ high glucose ＋ OZ 30,100,300 mmol/L）. The expression of p-TAK1, TAK1 binding protein （TAB1） , NF-KB p65 proteins of macrophages were analyzed by Western blot. The intracellular localization of NF-KB p65 was analyzed by cell im- mune fluorescence. The concentrations of TNF-α, IL-1β, MCP-1, Col IV and FN in each group cell media were determined by ELISA. Macrophages migration ability were observed microscope. Results Compared with control group, high glucose stimulate macrophage p-TAK1, TAB1, NF-KB p65 protein expression were significantly higher （P 〈 0. 05）. Both in eo-cuhureand monolayer cell culture, high glucose increases the concentration of TNF-α, IL- 1β, MCP-1, Col IV and FN （P 〈 0.05 ）. Exposed to high glucose, the concentration of TNF-α, IL-1β, MCP-1, Col IV and FN in co-cultured cells was higher than that in monolayer cells culture（P 〈 0. 05）. 5Z-7-oxozeaenol could decrease those cytokines secretion with a concentration dependent manner, comparing with high glucose group （P 〈 0. 05）. The number of macrophages migration were decreased by 300 nmol/L 5Z-7-oxozeaenol（P 〈 0. 05）. Con- clusion Exposed to high glucose, macrophages and mesangial cells can interact with each other to promote inflam- mation cytokines（TNF-α, IL-1β, MCP-1） and extracellular matrix components（Col IV, FN） secretion. TAK1 in- hibitor can reduce inflammation cytokinesand the secretion of extracellular matrix components by intervening NF-KB p65 nuclear transfer and inhibiting macrophage migration. TAK1 inhibitor plays a key role in anti-inflammation, an- ti

关 键 词：TAK1 共培养 糖尿病肾病 炎症 纤维化 

分 类 号：R587.24[医药卫生—内分泌]