血红素氧合酶-1基因修饰的骨髓间充质干细胞培养上清液对心肌梗死治疗作用的实验研究  

Experiment study of myocardial infarction treatment effect by heme oxygenase-1 gene-modified bone marrow mesenchymal stem cell culture supernatant

作　　者：王艾丽[1] 曾彬[2] 程新耀[1] 严斐斐[1] 

机构地区：[1]武汉大学中南医院心血管内科超声心动图室,武汉430071 [2]武汉大学人民医院心血管内科

出　　处：《中华临床医师杂志（电子版）》2013年第18期113-116,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(81270271)

摘　　要：目的探讨血红素氧合酶-1(HO-1)基因修饰的骨髓间充质干细胞(MSCs)培养上清液对大鼠心肌梗死的治疗作用。方法 HO-1腺病毒或对照GFP腺病毒转染MSCs,Western blot检测HO-1的表达;流式细胞仪检测无氧无血清条件下HO-1-MSCs、GFP-MSCs、MSCs的凋亡;ELISA和RT-PCR检测无氧无血清条件下HO-1-MSCs细胞因子的分泌。收集HO-1-MSCs、GFP-MSCs在无氧无血清条件下的培养上清液,于结扎冠状动脉1 h后多点注射到心肌梗死区边缘,注射Control Medium为对照组。注射4 d后检测心功能变化,4周后取梗死区边缘心肌行Masson染色和免疫组化CD34染色。结果 HO-1转染的MSCs可以稳定高效地表达HO-1蛋白(P=0.01),HO-1-MSCs在无氧无血清条件下的凋亡率显著低于GFP-MSCs和MSCs(P=0.01),且其在无氧无血清条件下HGF、bFGF、TGF-β、VEGF分泌水平显著增高(P=0.02);注射4 d后发现HO-1-MSCs上清液治疗组心功能各项指标均显著改善(P<0.01);4周后HO-1-MSCs上清液治疗组胶原面积明显减少(P<0.01),且梗死区微血管数量也增加。结论 HO-1-MSCs分泌多种细胞因子,通过促进血管新生,减少胶原沉积以及减少心肌坏死,从而改善心功能以及急性心肌梗死后的左心室重构。Objective To investigate the effect of the culture supernatants from MSCs overexpressing HO-1 .gene on a rat myocardial infarction. Methods MSCs were cultured in vitro and transfected by Adv-HO-1 or Adv-GFP. Western blot were used to determine the expression of HO-1. FACS was used to determined the apoptosis of MSCs under the condition of hypoxia plus serum-free. ELISA and RT-PCR were peformed to assay the expression and secrection of HGF, bFGF, TGF-[3 and VEGF. The culture supernatants from HO-1-MSCs or GFP-MSCs were injected into the infarcted border zone, separately. Four days after injection, cardiac function was measured by ultrasound, and 4 weeks after injection, the myocardial infarct size was measured with Masson＇s trichrome, and the CD34 in myocardial infarction area was detected by immunohistochemical method. Results HO-1-MSCs demonstrated HO-1 expression increase and were more resistant to hypoxia plus serum-free stimulation. Various growth factors, including HGF, bFGF, TGF-β and VEGF were produced by HO-1-MSCs, some of which were enhanced significantly under hypoxia plus serum-free stimulation. Four days after injection, the culture supernatants from HO-1-MSCs significantly improved the early cardiac function, and 4 weeks after injection, the culture supernatants from HO-1-MSCs significantly increased vascular regeneration, decreased the myocardium apoptosis, and decreased the collagen deposition. Conclusion HO-1-MSCs may improve cardiac function and left ventricular remodeling after acute myocardial infarction, which can secrete a variety of cytokines,promote angiogenesis, decrease collagen deposition and myocardial necrosis.

关 键 词：间质干细胞 心肌梗死 血红素加氧酶-1 细胞因子类 血管新生 

分 类 号：R542.22[医药卫生—心血管疾病]