血红素氧合酶-1修饰骨髓间充质干细胞对大鼠梗死心肌炎症细胞因子及心室重构的影响

Effects of mesenchymal stem cells modified by human heme oxygenase-1 gene on cardiac inflammatory cytokine and the ventricular remodeling

目的探讨移植血红素氧合酶-1（Heme Oxygenase，HO-1）修饰的骨髓间充质干细胞（mesenchymal stem cells，MSCs）对大鼠梗死心肌炎症细胞因子及心室重构的影响。方法取大鼠骨髓，扩增培养MSCs；HO-1或GFP腺病毒转染、DAPI标记MSCs。结扎大鼠左前降支制造心肌梗死模型1h后，分别将HO-1-MSCs、GFP-MSCs、PBS注射到大鼠心脏梗死区边缘，移植后第4天，Western Blot检测HO-1蛋白在梗死区边缘的表达，ELISA检测VEGF、bFGF、HGF的表达，RT-PCR检测炎症细胞因子TNF-α、IL-1β、IL-6、IL-10mRNA的表达。超声心动图检测大鼠心功能变化。移植4周后，梗死区心肌HE，Masson染色检测左室重构。结果HO-1-MSCs移植组与GFP-MSCs、PBS移植组相比，HO-1蛋白在梗死区边缘稳定高效表达；细胞因子VEGF、bFGF、HGF表达增加（P〈0．05）；炎症细胞因子TNF-α、IL-1β、IL-6mRNA表达减少（P〈0．05），而IL-10mRNA表达增加（P〈0．05）；左室舒张末期和收缩末期内径缩短，左室前壁增厚，心室腔减小，梗死区血管密度上调，胶原形成明显减少。结论HO-1-MSCs分泌多种生物活性细胞因子，协同HO-1蛋白改善炎症微环境，促进血管生成，减少胶原形成，从而延缓梗死区心室重构，有效改善心功能。

Objective To investigate the effects of mesenchymal stem cells（MSCs） transfected with human heme oxygenase-1 gene on the inflammatory cytokines and the ventricular remodeling after myocardial infarction in rats. Method MSCs were acquired from the bone marrow of adults rats. They were isolated, purified cultured, and transfected with Adv-HO-1, or Adv-GFP in vitro bofore transplantation. At 1 hours ＂after left coronary artery ligation, Adv-HO-1-MSCs or Adv-GFP-MSCs marked with DAPI were directly injected into the border of cardiac infarction in rats. At 4 days after transplantation, western blot analysis was used to measure HO-1 protein expression in the the border of cardiac infarction. The levels of VEGF, bFGF, HGF protein expression were measured by ELISA, and the levels of TNF-α、IL-1β, IL-6, IL-10 mRNA expression were measured by RT-PCR. The rat heart function was measured by echocardiography. At 4 weeks after transplantation, ventricular remodeling and pathological changes were measured by HE and Masson staining. Results The Adv-HO-1-MSCs treated group showed marked increase of HO-1 rotein （ P 〈 0.05）, and displayed significant increase of mentioned cytokines above, P 〈 0.05, compared with other groups. The Adv-HO-1-MSCs treated group displayed significant reduction of mRNAs expression of TNF-α、IL-1β, IL-6, and significant increase in IL-10 mRNA expression, with P 〈 0.05, compared with others. Conclusions HO-1-MSCs could secrete multiple eytokines in infarction hearts, and had beneficial effects on inflantmatory eytokines, remodeling processes and cardiac function.