Modulation of matrix metalloproteinase activity in human saphenous vein grafts using adenovirus-mediated gene transfer
Modulation of matrix metalloproteinase activity in human saphenous vein grafts using adenovirus-mediated gene transfer
复制标题
DOI：
10.1016/s0039-6060(98)70112-6
复制
发表时间：
1998-08-01
期刊：
影响因子：
3.8
通讯作者：
Galloway, AC
中科院分区：
文献类型：
--
作者：
Fernandez, HA;Kallenbach, K;Galloway, AC
背景人大隐静脉移植术后新生内膜的形成与多种基质金属蛋白酶（MMPs）及其组织抑制剂（TIMPs）有关。本研究评估了通过肾上腺病毒介导的基因转移调节hSVGs中MMP活性的可行性。首先，将1 × 10（9）噬斑形成单位（pfu）的编码β-半乳糖苷酶（Ad β gal）、MMP-3（AdMMP-3）或（TIMP-1）的复制缺陷型重组腺病毒加入hSVG的内腔中1小时。37 ℃孵育24 h后，采用免疫组化、原位酶谱和X-gal染色进行分析。AdMMP-3和AdTIMP-1感染的hSVGs的免疫组织化学将这些蛋白定位于内膜。原位酶谱显示，相对于AdTIMP-1或Ad β gal感染的血管，AdMMP-3感染的hSVGs内膜中MMP活性增加。MMP-3和TIMP活性可通过复制缺陷型重组腺病毒在hSVG中调节。我们以前已经证明，MMP-3或TIMP-1转导，或两者，抑制SMC迁移在体外重建血管壁。因此，MMP活性的调节可以在遗传工程改造的hSVG中提供高的通畅率。然而，腺病毒介导的基因传递仅限于血管内膜;需要开发感染中膜平滑肌细胞的策略。
Background. Neointima formation after human saphenous vein grafting (hSVG) involves several matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). This study assessed the feasibility of modulating MMP activity in hSVGs by adrenovirus-mediated gene transfer.Methods. First, 1 x 10(9) plaque-forming units (pfu) of replication-deficient recombinant adenoviruses encoding either beta-galactosidase (Ad beta gal), MMP-3 (AdMMP-3), or (TIMP-1) were added into the lumen of hSVGs for 1 hour. After incubation at 37 degrees C for 24 hours, specimens were analyzed by immunohistochemistry, in situ zymography, and X-gal staining.Results, By X-gal staining Ad beta gal-infected hSVGs stained positively in the intima and occasionally in the media. Immunohistochemistry of AdMMP-3- and AdTIMP-1-infected hSVGs localized these proteins to the intima. In situ zymography showed increased MMP activity in the intima of AdMMP-3-infected hSVGs relative to AdTIMP-1- or Ad beta gal-infected vessels.Conclusions. MMP-3 and TIMP activity can be regulated in hSVGs by replication-deficient recombinant adenoviruses. We have previously demonstrated that MMP-3 or TIMP-1 transduction, or both, inhibit SMC migration in an in vitro reconstituted vessel wall. Modulation of MMP activity may thus afford high patency rates in genetically engineered hSVGs. However, adenovirus-mediated gene delivery is limited to the vessel's intima; strategies to infect medial smooth muscle cells need to be developed.