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目的采用基因与细胞相结合的新方法,研究将经重组腺相关病毒(rAAV)介导的δ-SG基因修饰的骨髓间充质干细胞(MSCs)植入缺失δ-SG基因的TO-2型仓鼠心肌内,对其心功能的影响。方法将遗传性DCM鼠(TO-2仓鼠)分为MSCs组(n=15)、rAAV-δ-SG组(n=15)、rAAV-δ-SG-MSCs组(n=15),心室壁注入移植细胞。移植后5w和10w,分别进行心超检测左室舒张末直径(LVDd)、左室收缩末直径(LVSd),舒张末室间隔厚度(IVSd),舒张末左室后壁厚度(LVPW),左室射血分数(LVEF);免疫组化测心肌细胞特异蛋白;RT-PCR检测心肌中δ-SG基因表达;免疫荧光检测细胞分化、迁移;电镜检测新生细胞。结果与rAAV-δ-SG、MSCs组相比较,rAAV-δ-SG-MSCs组LVDd、LVSd减小(P<0.05),EF、IVSd、LVPW增大(P<0.05),而rAAV-δ-SG与MSCs组相比较无统计学差异;rAAV-δ-SG-MSCs组心肌细胞较成熟,结构较完整,心肌细胞特异蛋白表达增多,毛细血管密度增加(P<0.01);电镜中,可见结构完整的心肌细胞及新生血管出现。结论 rAAV-δ-SG-MSCs能与宿主细胞有效结合,与rAAV-δ-SG、MSCs治疗相比较,能更有效地修复TO-2仓鼠心肌细胞肌膜骨架蛋白,持续修复受损心肌,改善心功能,延缓心腔扩大。
OBJECTIVE: To study the effect of recombinant adeno-associated virus (rAAV) -mediated δ-SG gene modified bone marrow mesenchymal stem cells (MSCs) on TO-2 type Hamster myocardium, the impact on their cardiac function. Methods The hereditary DCM rats (TO-2 hamsters) were divided into MSCs group (n = 15), rAAV-δ-SG group Transplant the cells. The left ventricular end diastolic diameter (LVDd), left ventricular end systolic diameter (LVSd), left ventricular end diastolic dimension (IVSd), left ventricular end diastolic wall thickness (LVPW) (LVEF). Cardiomyocyte specific protein was detected by immunohistochemistry. Δ-SG gene expression in myocardium was detected by RT-PCR. Cell differentiation and migration were detected by immunofluorescence. Results Compared with rAAV-δ-SG and MSCs group, LVDd and LVSd in rAAV-δ-SG-MSCs group decreased (P <0.05), EF, IVSd and LVPW increased There was no significant difference between SG and MSCs group. The rAAV-δ-SG-MSCs group had more mature cardiomyocytes, more intact structure, more cardiomyocyte-specific protein and increased capillary density (P <0.01) Complete cardiomyocytes and neovascularization appear. Conclusions rAAV-δ-SG-MSCs can bind effectively to host cells. Compared with rAAV-δ-SG and MSCs, rAAV-δ-SG-MSCs could repair more effectively the sarcolemmal proteins of hamster ventricular cardiomyocytes of TO- Heart function, delayed heart chamber expansion.