TGF-β1参与全反式维甲酸对大鼠肾脏系膜细胞MMP-2表达的调节  

TGF-β1 Mediates the Regulation of the Expression of MMP-2 in Cultured Rat Mesangial Cell

作　　者：张农[1] 刘琛[1] 林文生[1] 张颂文[1] 顾建新[1] 郭慕依[1] 

机构地区：[1]复旦大学基础医学院病理学教研室,上海200032

出　　处：《复旦学报（医学版）》2001年第4期287-291,295,共6页Fudan University Journal of Medical Sciences

基　　金：上海市科技发展基金 (98QB14 0 0 7);卫生部优秀青年人才基金;CMB资助项目

摘　　要：目的　研究全反式维甲酸 (ATRA)对于大鼠肾脏系膜细胞表达MMP 2的影响以及转化生长因子 β1在这一过程中的作用。方法　采用Northernblot、Westernblot和Zymography法检测ATRA对培养大鼠肾脏系膜细胞MMP 2mRNA、蛋白分泌及酶活性表达变化 ;ATRA对系膜细胞表达TGF β1的作用 ,以及TGF β1对A TRA调节MMP 2表达的影响。结果　证实ATRA可增加培养大鼠肾脏系膜细胞的MMP 2mRNA表达、MMP 2蛋白分泌和增强MMP 2酶活性。ATRA也可引起TGF β1多肽呈剂量依赖型增加。反义TGF β1几乎可以完全阻断活性TGF - β1的分泌。经反义TGF β1转染的系膜细胞在 10 -6mol/LATRA作用下 ,其表达MMP 2mRNA的水平不发生变化 ,但是加入抗TGF β1抗体时 ,在 10 -6mol/LATRA作用下系膜细胞表达MMP 2呈剂量依赖型下降。结论　ATRA可上调培养大鼠系膜细胞的MMP 2mRNA、蛋白分泌和MMP 2酶活性的表达 ;TGF β1参与ATRA对MMP 2表达的调节 ,并为ATRA调节MMPPurpose: To study the regulatory effect of all-trans retinoic acid (ATRA) on the cultured rat mesangial cell expressing MMP-2 and the possible relationship between the transforming growth factor-β1 and the regulation of ATRA on the MMP-2 expression. Methods: The Northern blot, Western blot and Zymography assays were used to detect the expression of the mRNA, protein and zymatic activity of MMP-2 of cultured mesangial cell and the TGF-β1 transfectants. Results: ATRA upregulated the expression of MMP-2 mRNA and secretion of MMP-2 proteins, and increased enzymatic activity of MMP-2 in cultured mesangial cell. Besides, ATRA also increased a dose-dependent increase in the secretion of transforming growth factor-β1 (TGF-β1) peptides. Antisense TGF-β1 construct almost completely blocked secretion of active TGF-β1 peptides in the antisense- bearing transfectants. Northern blot analysis showed that the transfectants treated with 10-6 mol/L ATRA for 72 h neither increased nor decreased the levels of MMP-2 mRNA. Moreover, addition of anti-TGF-β1 antibodies to mesangial cell in the presence of 10-6 mol/L ATRA reduced ATRA-induced MMP-2 expression dose dependently. Conclusions: ATRA upregulated the expression of MMP-2 mRNA. ATRA also increased the secretion of MMP-2 protein and the enzymatic activity. TGF-β1 regulated the expression of MMP-2 in mesangial cell and it is necessary for the regulatory effect of ATRA. These data suggest that TGF-β1 might be involved in ATRA-induced MMP-2 expression.

关 键 词：转化生长因子-Β1 全反式维甲酸 基质金属蛋白酶-2 肾脏系膜细胞 ATRA MMP-2 

分 类 号：R364.3[医药卫生—病理学] R692.6[医药卫生—基础医学]