p38 MAPK在高糖诱导的肾小管上皮细胞转分化中的作用  

Effect of p38 MAPK on high glucose induced epithelial-mesenchymal-transition of human renal tubular epithelial cells

作　　者：吕智美[1] 邓华聪[1] 诸葛福媛[1] 刘金波[1] 冯正平[1] 陈丹燕[1] 南静[1] 

机构地区：[1]重庆医科大学附属第一医院内分泌科,重庆400016

出　　处：《第四军医大学学报》2008年第18期1641-1644,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30570414;30771038;30370670);教育部博士点基金(20050631009);教育部第二批"春晖计划";教育部重点项目(KJ050314);重庆市教委基金(渝教科[2003]7号文)

摘　　要：目的:探讨p38 MAPK在高糖诱导的肾小管上皮细胞转分化中的作用.方法:将体外培养的人肾小管上皮细胞(HK2)分为正常对照组(NG),高糖组(HG),无关siRNA转染组(RG)、转染p38 MAPK siRNA 24h组(24hG)、转染p38 MAPK siRNA 48h组(48hG).免疫细胞化学法检测α-平滑肌肌动蛋白(α-SMA)和细胞角蛋白(CK)的表达,RT-PCR法检测CK mRNA的表达,Western Blot检测p38 MAPK的活化及α-SMA的表达.结果:p38 MAPK siRNA能显著抑制高糖诱导的p38 MAPK过度活化;高糖刺激后肾小管上皮细胞α-SMA蛋白表达明显高于正常组(P<0.05),而p38 APK siRNA转染组其水平显著低于高糖组(P<0.05);高糖诱导HK2细胞CK mRNA和蛋白水平明显降低(P<0.05),而p38 MAPK siRNA转染后其水平显著高于高糖组(P<0.05);结论:高糖可导致肾小管上皮细胞p38 MAPK通路的激活而下调CK,同时导致α-SMA蛋白的表达,诱导其表型转化.AIM： To examine the roles of p38 MAPK in high glucose induced epithelial-mesenchymal-transition （EMT） in cultured human renal tubular epithelial cells （HK2）. METHODS： The cultured HK-2 cells were divided into 5 groups： normal glucose group （NG）, high glucose group （HG）, random siRNA group （RG）, 24h-p38 MAPK siRNA transfection group （24hG）, 48h-p38 MAPK siRNA transfection group （48hG）. The protein of a-smooth muscle actin （α-SMA） and eytokeratin （CK） was determined by immunoeytochemistry. RT-PCR was used to detect the mRNA expression of CK. Activity of p38 MAPK and expressions of α-SMA were observed by Western blot. RESULTS ： SiRNA for p38 MAPK inhibited the activation of p38 MAPK induced by high glucose. Upon the stimulation of high glucose, the expression of α-SMA protein significantly increased （ P 〈 0.05 ） , which was significantly decreased by transfection of p38 SiRNA （ P 〈 0.05 ）. Cultured with high glucose, the expression of CKm- RNA and protein decreased markedly （ P 〈 0.05 ）, but which was significantly increased by transfeetion of p38 SiRNA（ P 〈 0.05 ）. CONCLUSION： p38 MAPK may play an important role in high glucose-induced EMT characterized by gain of α-SMA expression and loss of CK expression in tubular epithelial cells.

关 键 词：P38丝裂原活化蛋白激酶类 肾小管 上皮细胞 上皮-间质转分化 

分 类 号：R587.1[医药卫生—内分泌]