纤维母细胞生长因子对肾脏足细胞增殖、骨形态蛋白-7以及细胞骨架蛋白表达的调节作用研究

›› 2008, Vol. 7 ›› Issue (5): 270-274.

纤维母细胞生长因子对肾脏足细胞增殖、骨形态蛋白-7以及细胞骨架蛋白表达的调节作用研究

王松税朝祥王尧尧阿拉塔张爱华王悦范敏华

100083 北京大学第三医院肾内科

收稿日期:2007-12-11 修回日期:1900-01-01 出版日期:2008-05-12 发布日期:2008-05-12
通讯作者: 王松

The regulation effects of fibroblast growth factor on podocyte proliferation and expressions of bone morphogenetic protein-7 and cytoskeletal proteins in podocytes

WANG Song, SHUI Chao-xiang, WANG Yao-yao, A La-ta, ZHANG Ai-hua, WANG Yue, WANG Tao, FAN Min-hua

Department of Nephrology, Peking University Third Hospital

Received:2007-12-11 Revised:1900-01-01 Online:2008-05-12 Published:2008-05-12
Contact: WANG Song

摘要/Abstract

摘要： 目的研究纤维母细胞生长因子(fibroblast growth factor，bFGF)对肾脏足细胞增殖、骨形态蛋白-7(bone morphogenetic protein-7，BMP-7)以及细胞骨架蛋白表达的调节作用。方法利用SV40肿瘤抗原转染所获得的永生性小鼠足细胞系培养，以集落形成法和噻唑蓝MTT还原实验测定不同浓度bFGF刺激下的足细胞增殖，采用SDS-聚丙烯酰胺凝胶电泳和Western blotting 检测BMP-7、alpha-平滑肌肌动蛋白(a-SMA)、以及波形蛋白(vimentin)的蛋白质水平表达，同时以甲基兰染色观察细胞形态。结果 bFGF(1.25～20 ng/ml)处理10天后明显增加足细胞集落生长，并呈现浓度依赖关系(r=0.844)，当bFGF达到20ng/ml时，集落数为对照培养的4.2倍，MTT实验亦获得一致的结果。甲基兰染色发现，bFGF刺激后的集落生长更加致密，直径增大，但同时出现集落形成细胞的空泡样改变。Western blotting 免疫印迹法检测到，bFGF(1.25～20ng/ml)处理96h后能显著抑制BMP-7的表达并呈浓度依赖关系，20ng/ml bFGF抑制＞90%。96h bFGF处理也抑制表达，但对vimentin表达无明显作用。结论以上结果表明bFGF刺激足细胞增殖的同时，对BMP-7的表达呈现下调作用，并引起空泡形成及α-SMA减少等细胞内部结构改变。基于BMP-7的肾保护作用，其降低可能是介导bFGF引起肾损伤的重要机制。

关键词: 纤维母细胞生长因子, 足细胞, 细胞骨架蛋白, 骨形态蛋白-7

Abstract: Objective To investigate the effect of fibroblast growth factor (bFGF) on podocyte proliferation and on expressions of bone morphogenetic protein-7 (BMP-7) and cytoskeletal proteins in podocytes. Method We employed a cell line of mouse podocytes immortalized with the simian virus 40 large tumor antigen gene, and examined the cell proliferation using colony-forming and MTT assays following treatment with various concentrations of bFGF. BMP-7, alpha-smooth muscle actin (e podocytes were detected by western blot. Methyl blue staining was used for the observation of cell morphology. Results After being treated with 1.25 to 20 ng/ml bFGF for 10 days, the podocytes showed significant increase of colony formation in a dose-dependent manner (r = 0.844). When bFGF in culture reached 20ng/ml, the number of colonies was 4.2 times higher than that of the controls. Similar results were obtained from MTT assay. On methyl blue staining, the colonies showed larger diameter and contained higher cell density after bFGF stimulation. A part of the cells also had vacuolation change. Western blot demonstrated that bFGF (1.25 to 20 ng/ml) treatment for 96 hours caused the decrease of BMP-7 in a dose-dependent manner, and that 20ng/ml bFGF stimulation resulted in the decrease of BMP-7 by >90%. bFGF treatment for 96 hours also inhibited Conclusion Our data suggest that bFGF induces podocyte proliferation and vacuolation, and down-regulates BMP-7 and

Key words: Podocyte, Cytoskeleton molecules, Bone morphogenetic protein-7

中图分类号:

R692

王松税朝祥王尧尧阿拉塔张爱华王悦范敏华. 纤维母细胞生长因子对肾脏足细胞增殖、骨形态蛋白-7以及细胞骨架蛋白表达的调节作用研究[J]. , 2008, 7(5): 270-274.

WANG Song;SHUI Chao-xiang;WANG Yao-yao;A La-ta;ZHANG Ai-hua;WANG Yue;WANG Tao;FAN Min-hua. The regulation effects of fibroblast growth factor on podocyte proliferation and expressions of bone morphogenetic protein-7 and cytoskeletal proteins in podocytes[J]. , 2008, 7(5): 270-274.