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作 者:邹敏书[1] 余健[1] 聂国明 何威逊[2] 丁冬胜[1] 李琳[1] 罗莉漫[1] 徐洪涛[1]
机构地区:[1]广州军区武汉总医院儿科,武汉430070 [2]上海交通大学附属上海儿童医院肾脏科,上海200040
出 处:《实用儿科临床杂志》2012年第5期351-353,共3页Journal of Applied Clinical Pediatrics
摘 要: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 influence of angiotensin Ⅱ(AngⅡ) or transforming growth factor-β1(TGF-β1) on the apoptosis of podocyte(PC) in vitro,whether bone morphogenetic protein-7(BMP-7) have protective effect on angiotensin Ⅱor TGF-β1 induce PC apoptosis and viability. Methods PC was preparated and carried out experiment as follows:1.PC was cultured with 10-8 mol·L-1,10-7 mol·L-1,10-6 mol·L-1 AngⅡor TGF-β1 for 24 h;2.PC was cultured with 10-7 mol·L-1 AngⅡor TGF-β1 for 12 h,24 h,36 h;3.PC was pretreated by 10-7 mol·L-1 BMP-7 and then exposed to 10-7 mol·L-1 AngⅡor TGF-β1 for 24 h.PC apoptosis and viability were measured by terminal-deoxynucleoitidyl transferase mediated TdT-mediated dUTP nick end labeling(TUNEL) staining and 3-(4,5-dimethylthiazol-zyl)-2,5-diphenyl thtrazolium bromide(MTT) methods,respectively.TGF-β1 in culture supernatants was detected by enzyme linked immunosorbent assay(ELISA) when PC was cultured with AngⅡ. Results With time length and concentrations of AngⅡ increased,AngⅡ induced PC apoptosis and the supernatants levels of TGF-β1 increased progressively,however PC viability decreased.TGF-β1 increased PC apoptosis in a time-and dose-dependent fashion and decreased PC viability.BMP-7 significantly ameliorated PC apoptosis,elevated PC viability,and decreased TGF-β1 level when PC was cultured with AngⅡ. Conclusion BMP-7 alleviate of AngⅡ induces PC apoptosis partly by decreased TGF-β1 level,and increase PC viability to protect against PC injury.
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