Zhu Tian-xiang, Lan Bin, Meng Ling-ying, Yang Yan-long, Li Rui-xiong, Li En-min, Zheng Shao-yi, Xu Li-yan
Department of Cardiovascular Surgery, Cardiovascular Institute, Affiliated Shantou Hospital of Sun Yat-sen University, Shantou, Guangdong Province 515031, China.
J Cardiothorac Surg. 2013 Jun 18;8:155. doi: 10.1186/1749-8090-8-155.
Currently, Saphenous vein (SV) and internal thoracic artery (ITA) are still the most common graft materials in Coronary Artery Bypass Grafting (CABG) whereas SV graft have a lower long-term patency than ITA. Vascular smooth muscle cells (VSMCs) phenotype conversion, proliferation and migration may play a key role in mechanism of vein graft restenosis. To explore differential gene expression profile in VSMCs from SV and ITA will help to further elucidate the mechanism of VSMCs in vein graft restenosis after CABG and to provide new thread of gene therapy.
VSMCs from paired SV and ITA were cultured for experiments of Affymetrix microarrays and verification using FQ RT-PCR, while the database for annotation, visualization and integrated discovery bioinformatics resources (DAVID 2.0) was utilized for bioinformatics analysis of differential gene expression profile between SV VSMCs and ITA VSMCs. RNA of tunica media from SV and ITA segments were extracted for FQ RT-PCR to display differential expression of PLAT RESULTS: 54,613 probe sets were examined by gene microarray experiments. In SV VSMCs, 1,075 genes were up-regulated and 406 of them were higher than two-fold; 1,399 genes were down-regulated and 424 of them were lower than two-fold as compare with ITA VSMCs.14 ECM-related genes differentially expressed were verificated and listed as following: COL4A4, COL11A1, FN1, TNC, THBS, FBLN, MMP3, MMP9, TIMP3, WNT5A, SGCD were higher whereas COL14A1, ELN, PLAT lower in SV VSMCs than ITA VSMCs. In addition, PLAT was lower in tunica media from SV segments than ITA.
VSMCs from SV and ITA have distinct phenotypes characteristics. Both promoting and inhibiting migration ECM-related genes were higher in VSMCs from SV as compared with ITA, suggesting that VSMCs from SV have more potential migrating capability whereas less PLAT both in SV VSMCs and vascular tissue,implying that SV may prone to be restenosis after CABG.
目前，大隐静脉（SV）和胸廓内动脉（ITA）仍是冠状动脉旁路移植术（CABG）中最常用的移植材料，然而大隐静脉移植的长期通畅率低于胸廓内动脉。血管平滑肌细胞（VSMC）表型转化、增殖和迁移可能在静脉移植物再狭窄机制中起关键作用。探索大隐静脉和胸廓内动脉VSMC中的差异基因表达谱将有助于进一步阐明冠状动脉旁路移植术后静脉移植物再狭窄中VSMC的机制，并为基因治疗提供新线索。
培养来自配对大隐静脉和胸廓内动脉的VSMC用于Affymetrix微阵列实验及荧光定量逆转录聚合酶链反应（FQ RT-PCR）验证，同时利用注释、可视化与整合发现生物信息学资源数据库（DAVID 2.0）对大隐静脉VSMC和胸廓内动脉VSMC之间的差异基因表达谱进行生物信息学分析。提取大隐静脉和胸廓内动脉段中膜的RNA进行FQ RT-PCR以显示组织型纤溶酶原激活物（PLAT）的差异表达。
通过基因微阵列实验检测了54,613个探针集。与胸廓内动脉VSMC相比，大隐静脉VSMC中1,075个基因上调，其中406个基因上调超过两倍；1,399个基因下调，其中424个基因下调超过两倍。验证了14个差异表达的细胞外基质（ECM）相关基因，如下所示：在大隐静脉VSMC中，Ⅳ型胶原α4链（COL4A4）、Ⅺ型胶原α1链（COL11A1）、纤连蛋白1（FN1）、肌腱蛋白C（TNC）、凝血酶敏感蛋白（THBS）、纤连蛋白（FBLN）、基质金属蛋白酶3（MMP3）、基质金属蛋白酶9（MMP9）、金属蛋白酶组织抑制因子3（TIMP3）、无翅型MMTV整合位点家族成员5A（WNT5A）、肌聚糖δ（SGCD）较高，而ⅩⅣ型胶原α1链（COL14A1）、弹力蛋白（ELN）、PLAT较低。此外，大隐静脉段中膜的PLAT低于胸廓内动脉。
大隐静脉和胸廓内动脉的VSMC具有不同的表型特征。与胸廓内动脉相比，大隐静脉VSMC中促进和抑制迁移的ECM相关基因均较高，提示大隐静脉VSMC具有更强的潜在迁移能力，而大隐静脉VSMC和血管组织中的PLAT均较少，这意味着冠状动脉旁路移植术后大隐静脉可能更容易发生再狭窄。
