去甲斑蝥素对TGF-β_1诱导的人近端肾小管细胞上皮细胞转分化及信号蛋白Smad2/3表达的影响  

Effect of norcantharidin on inhibition of tubular epithelial-mesenchymal transition and downregulation of Smad2/3 in HK-2 induced by TGF-β_1

作　　者：孙岩[1,2] 刘伏友[1] 孙林[1] 凌光辉[1] 李瑛[1] 刘健[2] 袁曙光[1] 刘虹[1] 张东山[1] 夏运成[1] 唐文彬[1] 

机构地区：[1]中南大学湘雅二医院肾内科,长沙410011 [2]新疆医科大学第一附属医院肾内科,乌鲁木齐830011

出　　处：《中南药学》2010年第3期170-174,共5页Central South Pharmacy

基　　金：国家中医药管理局中医药科学技术研究基金(06-07ZP33);湖南省卫生厅中医药科研基金(6103);湖南省科技计划资助项目(2007SK3040)

摘　　要：目的探讨去甲斑蝥素对体外人源性肾小管细胞株(HK-2)上皮细胞间质转分化(epithelial-mesenchymaltransition,EMT)以及信号蛋白Smad2/3的影响。方法体外培养HK-2细胞,分为对照组、TGF-β1组和去甲斑蝥素组(NCTD组)。对照组为无血清DMEM培养,TGF-β1组为TGF-β15 ng.mL-1诱导,NCTD组为不同浓度NCTD(0.5、1.0、2.5μg.mL-1)与TGF-β1(5 ng.mL-1)共同作用,各组作用时间48 h。使用半定量RT-PCR法检测α-平滑肌肌动蛋白(α-SMA)、上皮性钙黏附蛋白(E-cadherin)与Smad2、Smad3 mRNA表达;采用Western blot检测α-SMA、E-cadherin、Smad2/3、p-Smad2/3蛋白质表达。结果与对照组相比,TGF-β1组α-SMA mRNA及其蛋白表达明显上调(P<0.05),E-cadherin mRNA及其蛋白表达显著下调(P<0.05);与TGF-β1组相比,NCTD干预组α-SMA mRNA及其蛋白表达明显下调(P<0.05),而E-cadherin mRNA及其蛋白表达显著上调(P<0.05),且均呈剂量依赖性。与对照组相比,TGF-β1组Smad2、Smad3 mRNA和Smad2/3、p-Smad2/3蛋白表达上调(P<0.05);与TGF-β1组比较,NCTD干预组Smad2、Smad3 mRNA和Smad2/3、p-Smad2/3蛋白表达下调(P<0.05),具有剂量依赖关系。结论去甲斑蝥素具有抑制肾小管上皮细胞EMT的作用,该作用可能与其下调信号蛋白Smad2/3的表达有关。To investigate the effect of norcantharidin （NCTD） on epithelial-mesenchymal transition （EMT） and the expression of Smad2/3 in the HK2 cells induced by TGF-β1. Methods Human proximal tubular epithelial cells （HK-2） were divided into 3 groups： a control group （only FBS-free DMEM）, a TGF-β1 group （treated with TGF-β1 5 ng·mL^-1 ） , and an NCTD group （treated with TGF-β1 5 ng· mL^-1 and NCTD at different doses）. The expressions of α-SMA and E-cadherin in the HK-2 cells were determined by RT PCR and Western blot, respectively. The expressions of Smad2, Smad3 mRNA and Smad2/3, p-Smad2/3 protein in the HK-2 cells were also determined by RT- PCR and Western blot. Results The expression of α-SMA increased in the HK-2 induced by TGF-β1, while that of NCTD group treated by NCTD （0.5, 1.0, and 2.5 μg·mL^-1 ） was dramatically inhibited in a dose-dependent manner. The expression of E-cadherin in the HK-2 induced by TGF-β1 partly reversed by NCTD （0. 5, 1.0, and 2. 5μg·mL^-1 ） in a dose-dependent manner. The expressions of Srnad2, Smad3 mRNA and Smad2/3, p-Smad2/3 protein in the HK-2 induced by TGF-β1 （5 ng · mL^- 1 ） were suppressed by NCTD （0.5, 1.0, and 2.5μg·mL^-1 ） in a dose-dependent manner. Conclusion NCTD may inhibit EMT of tubular cells induced by TGF-β1, which may associate with the down-regulation of Smad2/3 expression.

关 键 词：去甲斑蝥素 上皮间质转分化 转化生长因子-Β1 SMAD2/3 

分 类 号：R966[医药卫生—药理学]