基质金属蛋白酶11对转化生长因子β1诱导人主动脉平滑肌细胞增殖和胶原合成的影响

Effect of MMP11 on the proliferation and collagen synthesis of human aortic vascular smooth muscle cells induced by TGF-β1

目的探讨基质金属蛋白酶(MMP)11对转化生长因子β1(TGF-β1)诱导的人主动脉(HA)-平滑肌细胞(VSMC)增殖和胶原合成的影响。方法将HA-VSMC分为对照组,0.1、1、5、10 ng/mL组(0.1、1、5、10 ng/mL TGF-β1)、5 ng/mL+si-NC组(5 ng/mL TGF-β1+转染si-NC)和5 ng/mL+si-MMP11组(5 ng/mL TGF-β1+转染si-MMP11),应用免疫印迹实验(Western blot)、MTS法、qPCR分别检测细胞周期蛋白D1(CyclinD1)、PCNA、胶原蛋白(Col)Ⅰ、ColⅢ和MMP11蛋白表达、细胞增殖和MMP11 mRNA表达。结果0.1、1、5和10 ng/mL浓度TGF-β1处理HA-VSMC后,0.1、1、5和10 ng/mL组HA-VSMC中CyclinD1、PCNA、ColⅠ、ColⅢ蛋白、MMP11蛋白、MMP11 mRNA水平及增殖活性均高于对照组(P<0.05)。5 ng/mL+si-MMP11组HA-VSMC的MMP11、CyclinD1、PCNA、ColⅠ、ColⅢ蛋白水平和增殖活性均低于5 ng/mL+si-NC组(P<0.05)。结论MMP11在不同浓度TGF-β1诱导的HA-VSMC中高表达,干扰MMP11能逆转TGF-β1诱导HA-VSMC增殖和胶原合成的作用。

Objective To investigate the effect of matrix metalloproteinase(MMP)11 on the proliferation and collagen synthesis of human aortic(HA)-vascular smooth muscle cells(VSMC)induced by transforming growth factorβ1(TGF-β1).Methods The experimental groups were as follows:the control group,0.1,1,5,10 ng/mL group(0.1,1,5,10 ng/mL TGF-β1),5 ng/mL+si-NC group(5 ng/mL TGF-β1+transfected si-NC)and 5 ng/mL+si-MMP11 group(5 ng/mL TGF-β1+transfected si-MMP11).Western blotting,MTS and RT-qPCR assay were conducted to detect the expression of cyclin D1(CyclinD1),PCNA,CollagenⅠ,CollagenⅢand MMP11,respectively.Results In 0.1,1,5,and 10 ng/mL TGF-β1-treated HA-VSMC,CyclinD1,PCNA,CollagenⅠ,CollagenⅢand MMP11 expression,as well as proliferation activity were higher than those in the control group(P<0.05,respectively).The protein level of CyclinD1,PCNA,CollagenⅠand CollagenⅢ,and proliferation activity in HA-VSMC in 5 ng/mL TGF-β1+si-MMP11 group were lower than those in 5 ng/mL+si-NC group(P<0.05,respectively).Conclusion MMP11 was highly expressed in HA-VSMC induced by TGF-β1 at different concentrations.Knock-down of MMP11 could reverse the effect of TGF-β1 on proliferation and collagen synthesis in HA-VSMC.