转化生长因子-β1通过活化ERK途径和上调Ets-1蛋白刺激基质金属蛋白酶-9产生  

Transforming growth factor-β1 stimulates matrix metalloproteinase-9 production through ERK activation pathway and upregulation of Ets-1 protein

作　　者：黄海长[1] 刘森炎[1] 梁燕[1] 刘颖[1] 李惊子[1] 王海燕[1] 

机构地区：[1]北京大学第一医院肾内科,100034

出　　处：《中华医学杂志》2005年第5期328-331,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目 ( 30270612 );教育部高校博士学科点专项科研基金资助项目(20020001070);"211工程 人类功能基因与疾病基因研究"资助项目

摘　　要：目的　探讨转化生长因子 -β1(TGF -β1)调节基质金属蛋白酶- 9(MMP -9)生成的分子机理。方法　选用小鼠肾小球足突细胞系,以细胞因子TGF -β1为刺激物,建立炎症细胞模型,分别应用明胶酶谱法和逆转录 PCR方法观察TGF -β1刺激细胞后培养上清MMP- 9活性及细胞MMP- 9mRNA变化;应用Western蛋白印迹检测TGF β1调节MMP-9中对ERK信号途径和转录因子Ets 1蛋白的影响。结果　对照组细胞上清有微弱MMP- 9活性, TGF β1刺激细胞 24h后培养上清MMP 9活性较对照组明显升高 (P<0 01),并呈TGF -β1刺激剂量依赖趋势; TGF- β1刺激 6h细胞MMP 9mRNA较对照组明显增加 (P<0 01),并且维持高水平至 24h;TGF- β1刺激细胞 4h转录因子Ets- 1蛋白增加(P<0 01),持续高水平至 24h。TGF- β1可以刺激细胞ERK-1 /2活化; ERK-1 /2活化阻断剂PD98059预处理细胞,可以阻断TGF β1刺激引起的Ets 1蛋白、MMP -9活性、MMP -9mRNA增加的效应。结论　以上结果提示TGF β1是通过活化细胞内ERK信号途径和上调转录因子Ets -1蛋白刺激足突细胞MMP- 9mRNA表达,继而蛋白活性增加。Objective To investigate the molecular mechanism of transforming growth factor β1 (TGF β1) in regulating the production of matrix metalloproteinase 9 (MMP 9) protein. Methods Mouse immortal podocyte cells were cultured. Different concentrations (1, 2, and 5 ng/l) of TGF β1 were added into the culture mediom. Cell culture without TGF β1 stimulation was used as control group. The activity of MMP 9 in the supernatant of the culture medion was assayed by gelatin zymography, expression of MMP 9 mRNA was assessed by RT PCR; the activation of ERK pathway and the level of a transcriptional factor Ets 1 protein was analyzed by Western blotting. PD98059, a specific inhibitor of ERK1/2 activation, was added into the culture fluid of the podocytes for 30 minutes, than 2ng/ml TGF β1 was added. The above mentioned tested were conducted to observe the influence of the inhibitor of ERK1/2 activation. Results The MMP 9 activity was very week in the supernatant of culture fluid of the control group and was increased in the TGF β1 groups dose dependently. After the podocytes were co incubated with 1 ng/ml TGF β1 for 24 hours, the MMP 9 activity was 26 86 times that of the control group ( P <0 01). Since the 12 th hour after co incubation with 2ng/ml TGF β1 the MMP 9 activity in the supernatant of culture fluid began to be significantly increased and remained at high level till the 48 th hour. RT PCR showed that low level MMP 9 mRNA expression in the control group. After stimulation of 2 ng/ml TGF β1 for 6 hours the MMP 9 mRNA expression was 2 71 times that of the control group ( P <0 01) and the high level expression lasted 24 hours. Western blotting showed low level Ets 1 protein in the control group. At the time point of 12 th hour after stimulation of TGF β1 the Ets 1 protein expression was increased in all the three TGF β1 groups. After stimulation with 2ng/ml TGF β1 for 4 hours the Ets 1 protein expression was 2 71 times that of the control group ( P <0 01). After pr

关 键 词：TGF-β1 MMP-9 刺激细胞 ETS-1 ERK 对照组 转化生长因子-Β1 转录因子 白刺 信号途径 

分 类 号：R692.6[医药卫生—泌尿科学]