联合应用粒细胞集落刺激因子和骨髓基质干细胞治疗急性心肌缺血的实验研究

Autologous bone marrow stromal cell transplantation with application of granulocyte colony-stimulating factor improves rabbit cardiac performance after actue myocardial infarction

目的探讨粒细胞集落刺激因子和自体骨髓基质干细胞(MSCs)移植共同应用对心肌缺血的治疗作用。方法采用MSCs在体外培养扩增。在结扎冠状动脉造成急性心肌缺血后,把被5溴-2脱氧尿苷(BrdU)标记后的MSCs移植到自体的缺血心肌中,同时腹腔注射粒细胞集落刺激因子5d。4周后通过激光共聚焦显微镜验证移植后的MSCs是否向心肌细胞分化,通过心脏彩超和多导生理记录仪测定缺血心肌在移植自体MSCs后是否能提高心功能。另外,用Masson氏三色染色法从左室中断面切片量化心肌梗死范围。结果移植4周后,MSCs向心肌细胞分化,表达出α-横纹肌肌动蛋白和存在于心肌闰盘中的connexin43,联合应用粒细胞集落刺激因子和MSCs的治疗组左心室收缩功能恢复较好,心肌梗死面积最小,单独应用MSCs移植的治疗组次之,对照组心肌左室收缩功能恢复较差,心肌梗死面积最大。结论MSCs移植后可以向心肌细胞分化,粒细胞集落刺激因子可以动员机体内自体MSCs到心肌缺血区域,二者联合应用对于缺血心肌有协同治疗作用。

Objective To test the effect of autologus marrow stromal cells （MSCs） transplantation combined with application ofgranulocyte colony stimulating factor （G-CSF） on the function ofischemic hearts in vivo. Methods Acute myocardial infarction was induced in rabbits by occlusion of the left anterior descending artery, and autologous MSCs labeled with BrdU in vitro was introduced in the infarct area of the same donor rabbit. G-CSF was administered by subcutaneous injection for 5 consecutive days. Four weeks later, the transplanted MSCs were detected by laser scanning confocal microscopy and the cardiac function of the rabbits was examined by echocardiogram and multichannel physiological recorder. The myocardial infarct size was measured on the mid-transverse sections stained with Masson＇s trichrome. Results Four weeks after transplantation, the MSCs were found to undergo myogenic differentiation with expressions of α-sarcomeric actin and connexin 43 in the intercalated disk. MSC transplantation in combination with G-CSF administration improved the left ventricular contractility and markedly reduced myocardial infarct size as compared with cell transplantation without G-CSF. Conclusion Application of G-CSF following autologous MSC transplantation may represent a promising therapeutic strategy for ischemic heart disease.