干预Hedgehog信号对肾小管上皮细胞增殖细胞核抗原表达的影响  

Intervene of Hedgehog signal on expression of proliferating cell nuclear antigen in renal tubular epithelial cells

作　　者：陆红[1] 吴莲凤[1] 林成成[2] 王斯璐[2] 梁勇[2] 白永恒[2] 

机构地区：[1]温州医科大学附属第一医院医学检验中心,浙江温州325015 [2]温州医科大学附属第一医院外科实验室,浙江温州325015

出　　处：《温州医学院学报》2015年第1期14-17,共4页Journal of Wenzhou Medical College

基　　金：浙江省自然科学基金资助项目(LQ12H05001);温州市科技计划项目(Y20110028;Y20130158)

摘　　要：目的:探讨Hedgehog(HH)信号通路的活化和抑制对肾小管上皮细胞中增殖细胞核抗原(PCNA)表达的影响。方法:根据干预措施将体外培养的大鼠肾小管上皮细胞NRK-52E分为对照组、活化组(10μg/L Shh)和干预组(10μg/L Shh加5μmol/L环杷明)。细胞培养24 h后,细胞免疫荧光染色检测HH信号关键分子Ptch1、Smo和Gli1,以及细胞增殖标志物PCNA的表达,real-time RT-PCR检测Ptch1和Smo m RNA的表达。结果:免疫荧光染色显示,对照组细胞维持一定的增殖状态,HH信号活性较低,PCNA表达水平较低;当外源性重组蛋白Shh活化NRK-52E细胞24 h后,Smo和Gli1 m RNA和蛋白表达显著升高,Ptch1的表达显著下调,这提示Shh可激活HH信号(P<0.05)。在此过程中,PCNA表达水平显著升高,NRK-52E细胞数量增加,同时伴随细胞形态的改变;在Shh的基础上加入环杷明干预后,HH信号被抑制(P<0.05),PCNA表达下降,NRK-52E细胞增殖被抑制,出现凋亡或坏死。结论:HH信号与肾小管上皮细胞的增殖密切相关。通过药物调控HH信号的活化,可影响PCNA的表达,进而影响细胞增殖效应。Objective： To explore and investigate the activation and inhibition of Hedgehog signal regulated on expression of proliferating cell nuclear antigen （PCNA） in renal tubular epithelial cells （NRK-52E）.Methods：NRK-52E cells were randomly divided into control group （treated with solvent only）, activated group （treated with Shh at the concentrations with 10 μg/L） and treatment group （treated with Shh at the concentration of 10 μg/L plus cyclopamine at the concentrations with 5 μmol/L）. After cultured for 24 h, immunolfuorescence staining was used to evaluate the expressions of Ptch1, Smo, Gli1, and PCNA. The expression levels of Ptch1 and Smo mRNA were quantiifed by real-time RT-PCR.Results： NRK-52E cells without drug treatment maintain slowed proliferation state, and HH signal was not activated, and PCNA was in low expression. Treatment with exogenous recombinant protein Shh for 24 h, the mRNA or protein expression levels of Smo and Gli1 were signiifcantly in-creased, and the expression of Ptch1 was decreased, suggesting that Shh activates the HH signal （P〈0.05）. In this process, the expression of PCNA protein was signiifcantly increased. NRK-52E cells showed proliferation, and morphologic change occurred. After cyclopamine intervention, the activity of HH signal was decreased （P〈0.05）, and PCNA expression was also inhibited. In addition, abnormal proliferation of NRK-52E cells was inhibited, and increased apoptosis and necrosis cells.Conclusion： HH signal is involved in the proliferation of renal tubular epithelial cell. Regulating the activation of HH signal inlfuences PCNA expression and cell proliferation.

关 键 词：Hedgehog信号 环杷明 肾小管上皮细胞 增殖细胞核抗原 

分 类 号：R692[医药卫生—泌尿科学]