血管内皮生长因子调节小鼠系膜细胞基质金属蛋白酶及其组织抑制物的表达

VEGF regldates the expression of MMPs and TIMPs in mouse mesangial cells

目的 研究血管内皮生长因子(VEGF)对体外培养的小鼠系膜细胞表达基质金属蛋白酶(MMPs)和金属蛋白酶组织抑制物(TIMPs)的调节作用。方法 对体外培养的小鼠系膜细胞应用人重组VEGF孵育12 h,应用蛋白质印迹法检测细胞培养液MMP2、MMP9、TIMP1、TIMP2的蛋白质水平;应用白明胶酶谱法检测MMP2、MMP9的活性;应用RT-PCR检测系膜细胞TIMP1、TIMP2 mRNA表达。结果 VEGF(10 ng/ml)可提高小鼠系膜细胞MMP2和MMP9蛋白质释放(和对照组相比分别提高43％和34％,P<0.05),MMP2和MMP9活性分别提高17％和26％(P<0.05)。同时,VEGF可呈剂量依赖地下调小鼠系膜细胞TIMP1和TIMP2蛋白质和mRNA的表达,VEGF(25 ng/ml)可减少小鼠系膜细胞TIMP1和TIMP2蛋白质释放分别为43％和67％(P<0.05);并抑制TIMP1和TIMP2mRNA的表达(分别下降41％和59％,P<0.01)。结论 VEGF使系膜细胞MMPs蛋白质表达增加、活性增强,同时下调TIMPs mRNA和蛋白质的表达,可能在增生性肾小球肾炎的发病机制中起一定的作用。

Objective To study the regulation of MMPs and TIMPs expression by VEGF in mouse mesangial cells (MMCs) . Methods Cultured MMCs were incubated with or without recombinant human VEGF for 12 hours. Protein levels of MMP2,MMP9,TIMP1 and TIMP2 in media were measured by Western blot analysis. MMP2 and MMP9 activities were measured by gelatin zymography. TIMP1 and TIMP2 mRNA expression in MMCs was analyzed by RT-PCR. Results VEGF(10 ng/ml) upregulated MMP2 and MMP9 protein secretion by MMCs (increased by 43% and 34% respectively, P <0. 05) when compared with control. MMP2 and MMP9 activities also increased by 17% and 26% ( P < 0. 05) respectively. Simultaneously, VEGF downregulated TIMP1 and TTMP2 protein secretion and mRNA expression by MMCs in a dose-dependent manner. VEGF(25 ng/ml) decreased TIMP1 and TIMP2 protein secretion by 43% and 67% (P < 0. 05) and suppressed TIMP1 and TIMP2 mRNA expression by 41% and 59% , respectively ( P < 0. 01) by MMCs. Conclusion VEGF increases MMPs excretion and activity but suppresses TIMPs protein and mRNA expression by MMCs, which may promote cell proliferation and ECM accumulation in proliferative glomerulonephritis.