肿瘤坏死因子-α对原代大鼠肾近端小管上皮细胞Toll样受体2表达的调控及机制  

Modulation of the protein expression of TLR2 by TNF-α in primary rat renal proximal tubule cells

作　　者：刘珊英[1] 李焱[1] 常建星[1] 范新兰[1] 黄琼[1] 傅玉如[1] 林燕华[1] 林天歆[1] 

机构地区：[1]中山大学附属第二医院医学研究中心,广东广州510120

出　　处：《中国药理学通报》2009年第10期1292-1295,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30671974);广东省自然科学基金资助项目(No4009408);广东省医学科学基金资助项目(No2004197)

摘　　要：目的探讨肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）对原代大鼠肾近端小管上皮细胞Toll样受体2（tolllike receptor2,TLR2）表达的调控以及核因子-κB（nuclearfactor-κB,NF-κB）在其中的作用。方法体外分离与培养原代大鼠肾近端小管上皮细胞,以TNF-α按不同时间给予刺激,Western blot检测TLR2,I-κBα,磷酸化I-κBα（pI-κBα）,GAPDH蛋白水平。分别以TNF-α、NF-κB特异性抑制剂Bay11-7082处理25h、Bay11-7082预处理1h后加入TNF-α刺激24h,检测TLR2表达的变化。结果TNF-α刺激6~24h,TLR2高于基线水平;Bay11-7082处理组和Bay11-7082＋TNF-α处理组的TLR2蛋白表达水平高于对照组。Bay11-7082＋TNF-α处理组与单独TNF-α处理组TLR2蛋白表达水平无差异。TNF-α刺激后5~120min,pI-κBα蛋白水平升高。结论TNF-α促进原代大鼠肾近端小管上皮细胞TLR2蛋白表达,NF-κB对TLR2的表达可能起负调节作用。Aim To investigate the modulation of the expression of toll like receptor 2 （TLR2） in primary rat renal proximal tubule cells by tumor necrosis factor- α （TNF-α） and the implication of nuclear factor-κB （NF-κB） in the process. Methods Rat renal proximal tubule cells（PTCs） isolated from male adult Sprague Dawley rats were treated with TNF-α for the indicated time courses. A specific NF-κB inhibitor, Bayl 1- 7082, was used alone or as a pretreatment for 1 h followed by exposure to TNF-α for 24 h. The protein lev- els of TLR2, I-κBα, phosphorylated I-κBα,and GAP- DH were detected through Western blot with specific antibodies. Results The protein level of TLR2 was significantly increased in the presence of TNF-α for 6, 12,and 24 h. Treatment with Bayll-7082 for 25 h a-lone or as a pretreatment for 1 h followed by addition of TNF-α for 24 h resulted in a remarkable increase in TLR2 as compared with the controls. No significant difference in TLR2 protein level was detected between TNF-α group and Bayll-7082 plus TNF-α treated group. An increase in phosphorylated I-κBα was observed from 5 to 120 min and a derease in total I-κBα was observed by 30 min after treatment with TNF-α at a dose of 10 μg ·L^-1 in PTCs. Conclusion TNF-α induces an upregulation of TLR2 protein level independent of NF-κB in primary rat renal proximal tubule cells. NF-κB may serve as a negative regulator for TLR2 expression in rat PTCs.

关 键 词：肿瘤坏死因子-Α TOLL样受体2 核因子-ΚB κB抑制物 Bay11-7082 肾脏近端小管上皮细胞 

分 类 号：R-332[医药卫生] R322.61