过表达BMP7对TGF-β1诱导肾小管上皮细胞功能的影响

Effect of BMP7 over-expression on the function of RTEC induced by TGF-β1

目的通过对大鼠肾小管上皮细胞(RTEC)转染外源性骨形态发生蛋白7(BMP7)基因,探讨BMP7对RTEC增殖活性和迁移能力的影响。方法分别在体外用腺病毒介导载体HBAD-GFP和HBADBMP7转染入大鼠RTEC,转化生长因子-β1(TGF-β1)刺激RTEC,反转录PCR(RT-PCR)及Western blot方法检测其mRNA及蛋白表达,MTS和迁移实验检测RTEC增殖及迁移能力的变化。结果 (1)成功建立转染细胞系,TGF-β1在10 ng/m L作用12 h时促RTEC增殖、抑制BMP7的表达效果最佳;(2)HBAD-BMP7转染RTEC后,BMP7的mRNA和蛋白水平均明显增高(P<0.01),p38的mRNA和蛋白水平明显下降(P<0.01)。(3)RTEC过表达BMP7后明显缓解TGF-β1的促增殖和迁移能力,差异有统计学意义(P<0.05)。结论过表达BMP7可降低p38表达,改善TGF-β1诱导RTEC的功能。

Objective To demonstrate the effects of exogenous bone morphogenetic protein 7(BMP7） overexpression on proliferation activity and migration ability of transforming growth factor β1(TGF-β1）-induced rat renal tubular epithelial cells(RTEC）. Methods HBAD-GFP and HBAD-BMP7 were transfected into rat RTEC using adenovirus-mediated vectors in vitro. RT-PCR and Western blot were used to detect the expression of BMP7 mRNA and protein.MTS and migration test were used to detect the proliferation and migration abilities of RTEC. Results The transfected cell line was successfully established,and TGF-β1 promoted the proliferation of RTEC and inhibited the expression of BMP7. The most striking effect was obtained under the concentration of TGF-β1 of 10 ng/m L at 12 h. After HBAD-BMP7 transfection with RTEC,the mRNA and protein levels of BMP7 were significantly increased(P〈 0. 01）. Moreover,the mRNA and protein levels of p38 significantly decreased(P〈 0. 01）. After over-expression of BMP7,RTEC significantly alleviated the TGF-β1-induced proliferation and migration abilities(P〈 0. 05）. Conclusion High expression of BMP7 can improve the function of TGF-β1-induced rat RTEC by reducing p38 pathway.