Turner Neil A, Hall Kersten T, Ball Stephen G, Porter Karen E
Institute for Cardiovascular Research, School of Medicine, Worsley Building, University of Leeds, Leeds LS2 9JT, UK.
Atherosclerosis. 2007 Jul;193(1):36-43. doi: 10.1016/j.atherosclerosis.2006.08.017. Epub 2006 Sep 18.
Activation of matrix metalloproteinases (MMPs) facilitates smooth muscle cell (SMC) invasion, an important event in the development of intimal hyperplasia in saphenous vein (SV) bypass grafts. In this study, we performed selective gene silencing using small inhibitory RNA (siRNA) oligonucleotides to examine the relative contributions of MMP-2 and MMP-9 to the invasiveness of cultured human SV-SMCs. Cultures were established from human SV obtained from patients undergoing coronary artery bypass surgery. Transfection of SV-SMCs with MMP-2 siRNA selectively reduced MMP-2 secretion and inhibited invasion through a Matrigel barrier. Supplementation of medium with recombinant MMP-2 overcame these effects. Similarly, transfection of SV-SMCs with MMP-9 siRNA selectively reduced MMP-9 secretion and subsequent invasion, effects reversed by recombinant MMP-9 supplementation. Neither MMP-2 nor MMP-9 siRNA inhibited SV-SMC migration in the absence of a Matrigel barrier. Our data demonstrate that selective gene-silencing of either MMP-2 or MMP-9 markedly reduces the invasive capacity of cultured human SV-SMCs, indicating that these MMPs play distinct non-overlapping roles in SV-SMC invasion in vitro. Specific manipulation of either MMP-2 or MMP-9 may therefore provide a valuable strategy for prevention of SV graft stenosis in man.
基质金属蛋白酶(MMPs)的激活促进平滑肌细胞(SMC)侵袭,这是大隐静脉(SV)旁路移植内膜增生发展过程中的一个重要事件。在本研究中,我们使用小干扰RNA(siRNA)寡核苷酸进行选择性基因沉默,以研究MMP - 2和MMP - 9对培养的人SV - SMC侵袭性的相对贡献。培养物来自接受冠状动脉搭桥手术患者的人SV。用MMP - 2 siRNA转染SV - SMC可选择性降低MMP - 2分泌,并抑制其通过基质胶屏障的侵袭。用重组MMP - 2补充培养基可克服这些影响。同样,用MMP - 9 siRNA转染SV - SMC可选择性降低MMP - 9分泌及随后的侵袭,补充重组MMP - 9可逆转这些影响。在没有基质胶屏障的情况下,MMP - 2和MMP - 9 siRNA均未抑制SV - SMC迁移。我们的数据表明,对MMP - 2或MMP - 9进行选择性基因沉默可显著降低培养的人SV - SMC的侵袭能力,表明这些MMPs在体外SV - SMC侵袭中发挥着不同的、不重叠的作用。因此,对MMP - 2或MMP - 9进行特异性调控可能为预防人类SV移植物狭窄提供一种有价值的策略。