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目的探讨血管紧张素Ⅱ(AngⅡ)、转化生长因子-β1(TGF-β1)对体外足细胞(PC)凋亡的影响,及骨形成发生蛋白-7(BMP-7)对AngⅡ或TGF-β1介导PC凋亡及活力是否有保护作用。方法制备PC,进行如下实验:1.PC与10-8mol.L-1、10-7mol.L-1、10-6mol.L-1AngⅡ或TGF-β1培养24 h;2.PC分别与10-7mol.L-1的AngⅡ或TGF-β1培养12 h、24 h、36 h;3.BMP-710-7mol.L-1预处理PC,再与10-7mol.L-1AngⅡ或TGF-β1一起培养24 h。末端脱氧核糖核苷酸转移酶介导的dUTP切口末端标记技术(TUNEL)法检测PC的凋亡,四甲基偶氮唑蓝(MTT)测定PC活力,ELISA测定PC和AngⅡ一起培养时上清液中TGF-β1水平。结果随AngⅡ作用时间延长及浓度增加,PC凋亡愈明显,PC活力降低,上清液中TGF-β1水平亦升高。TGF-β1亦以剂量时间依赖性增加PC凋亡,降低PC活力。BMP-7减少PC凋亡,增加PC活力,并降低PC和AngⅡ一起培养的上清液中TGF-β1水平。结论 BMP-7可下调TGF-β1水平,减轻AngⅡ诱导PC凋亡,增加PC活力,对PC损伤有保护作用。
Objective To investigate the effects of angiotensin Ⅱ (AngⅡ) and transforming growth factor-β1 (TGF-β1) on the apoptosis of podocytes in vitro and the effect of BMP-7 on AngⅡ or TGF-β1 Mediated PC apoptosis and vitality whether there is a protective effect. Methods PC were prepared for the following experiments: 1.PC was incubated with 10-8mol.L-1, 10-7mol.L-1, 10-6mol.L-1AngⅡ or TGF-β1 for 24 h. 7mmol.L-1 AngⅡ or TGF-β1 for 12 h, 24 h, 36 h; 3. Pretreatment of PC with BMP-710-7 mol.L-1 followed by 10-7 mol·L-1AngⅡ or TGF-β1 Cultivation 24 h. The apoptosis of PC was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), the activity of PC was determined by MTT, and the supernatant Fluid TGF-β1 levels. Results With the prolongation of Ang Ⅱ and the increase of concentration, the apoptosis of PC was more obvious, the activity of PC decreased, and the level of TGF-β1 in supernatant also increased. TGF-β1 also increased PC apoptosis in a dose-and time-dependent manner and decreased PC viability. BMP-7 decreases PC apoptosis, increases PC viability, and decreases TGF-β1 levels in supernatants cultured with PC and Ang II. Conclusion BMP-7 can down-regulate the level of TGF-β1, reduce the apoptosis of PC induced by AngⅡ, increase the activity of PC, and protect PC from injury.