Cheng L, Mantile G, Pauly R, Nater C, Felici A, Monticone R, Bilato C, Gluzband Y A, Crow M T, Stetler-Stevenson W, Capogrossi M C
Laboratory of Cardiovascular Science, National Institute on Aging, National Institutes of Health, Bethesda, MD, USA.
Circulation. 1998 Nov 17;98(20):2195-201. doi: 10.1161/01.cir.98.20.2195.
Endovascular injury induced by balloon withdrawal leads to the increased activation of matrix metalloproteinases (MMPs) in the vascular wall, allowing smooth muscle cells (SMCs) to digest the surrounding extracellular matrix (ECM) and migrate from the media into the intima. The objective of this study was to examine the effects of a replication-deficient adenovirus carrying the cDNA for human tissue inhibitor of metalloproteinase-2 (AdCMV.hTIMP-2) on SMC function in vitro and neointimal development in the injured rat carotid artery.
Infection of cultured rat aortic SMCs at a multiplicity of infection of 100 with AdCMV.hTIMP-2 resulted in high-level expression of hTIMP-2 mRNA and protein secretion into the medium. Conditioned media (CM) from AdCMV. hTIMP-2-infected but not control virus (AdCMV.null or AdCMV. betagal)-infected SMCs inhibited MMP-2 activity on gelatin zymograms as well as the chemoattractant-directed migration of SMCs across reconstituted basement membrane proteins in the Boyden chamber assay. In contrast, AdCMV.hTIMP-2 CM had no effect on chemoattractant-directed migration of SMCs occurring in the absence of an ECM barrier or on the proliferation of cultured neointimal SMCs. Delivery of AdCMV.hTIMP-2 (2.5x10(9) pfu) to the carotid artery wall at the time of balloon withdrawal injury inhibited SMC migration into the intima by 36% (P<0.05) at 4 days and neointimal area by 53% (P<0.01) at 8 days and by 12% (P=NS) at 21 days after injury. AdCMV.hTIMP-2 had no effect on medial area.
Adenovirus-mediated hTIMP-2 gene transfer inhibits SMC invasiveness in vitro and in vivo and delays neointimal development after carotid injury.
球囊回撤引起的血管内损伤导致血管壁中基质金属蛋白酶（MMPs）的激活增加，使平滑肌细胞（SMCs）能够消化周围的细胞外基质（ECM）并从中膜迁移至内膜。本研究的目的是检测携带人金属蛋白酶组织抑制剂-2（hTIMP-2）cDNA的复制缺陷型腺病毒对体外平滑肌细胞功能以及损伤大鼠颈动脉新生内膜形成的影响。
用AdCMV.hTIMP-2以感染复数100感染培养的大鼠主动脉平滑肌细胞，导致hTIMP-2 mRNA高水平表达并分泌蛋白质至培养基中。来自AdCMV.hTIMP-2感染而非对照病毒（AdCMV.null或AdCMV.betagal）感染的平滑肌细胞的条件培养基（CM）在明胶酶谱上抑制MMP-2活性，并且在Boyden小室试验中抑制趋化因子引导下平滑肌细胞穿过重组基底膜蛋白的迁移。相比之下，AdCMV.hTIMP-2 CM对在没有ECM屏障情况下发生的趋化因子引导下的平滑肌细胞迁移或对培养的新生内膜平滑肌细胞的增殖没有影响。在球囊回撤损伤时将AdCMV.hTIMP-2（2.5×10⁹ pfu）递送至颈动脉壁，在损伤后4天抑制平滑肌细胞向内膜的迁移达36%（P<0.05），在8天抑制新生内膜面积达53%（P<0.01），在21天抑制12%（P=无显著性差异）。AdCMV.hTIMP-2对中膜面积没有影响。
腺病毒介导的hTIMP-2基因转移在体外和体内均抑制平滑肌细胞侵袭，并延迟颈动脉损伤后的新生内膜形成。
