大鼠脂肪干细胞联合碱性成纤维生长因子治疗大鼠心肌梗死

目的 探讨脂肪干细胞（ADSC）联合碱性成纤维生长因子（bFGF）提高心肌梗死大鼠心脏功能的可能机制.方法 SD大鼠50只,先行脂肪干细胞提取手术,分离、培养、鉴定脂肪干细胞,行干细胞成脂、成骨诱导分化,建立大鼠心肌梗死模型,选择缩短分数（FS）＜40％的SD大鼠（36只）,完全随机分4组：单纯注射PBS组（n=9）、单纯注射bFGF组（n=9）、PBS＋ADSC组（n=9）以及ADSC＋bFGF组（n=9）,以bFGF胶为可注射性载体,ADSC为种子细胞,1周后进行大鼠心梗部位移植.移植4周后对心梗面积、左心室室壁厚度、心梗部位微血管密度、ADSC在心梗部位分化情况等指标进行检测,并利用心脏超声对心梗大鼠的心脏功能进行测定.应用免疫组织化学、免疫荧光等方法评价其改善心脏功能可能机制.结果 成功分离ADSC并且通过流式细胞术表明大多数分离的脂肪干细胞CD90抗体阳性、CD29抗体阳性、CD45抗体和CD34抗体阴性.脂肪干细胞能够经过诱导分化成成脂、成骨细胞.PBS＋ADSC组以及ADSC＋bFGF组在室壁厚度、梗死面积、梗死区域微血管形成能力与PBS组相比,差异均有统计学意义,且以ADSC＋bFGF组最为显著（P＜0.01）.ADSC移植到大鼠心脏4周后,可以分化为cTnT阳性细胞、SMA阳性细胞以及vWAg阳性细胞.心功能检测结果表明PBS＋ADSC组同ADSC＋bFGF组一样,具有心梗修复能力.结论 移植ADSC的同时增加bFGF能促进梗死区的心肌再生以及血管化,并能显著改善心脏功能.

Treatment of myocardial infarction by co-injection of rat adipose derived stem cell with basic fibroblast growth factor

Objective To This project aimed to discover potential mechanism of treating myocardial infarction by transplantation of adipose-derived stem cells （ADSC） with basic fibroblast growth factor（bFGF）.Methods ADSC were isolated from the fat tissue.These cells were cultured and induced to differentiate to the adipogenic cells,which were identified by rows stem cell adipogenic and osteogenic differentiation.Myocardial infarction （MI） model were conducted in 50 SD rats,and rats were randomized into 4 groups with the inclusion criterion of EF （ejection fraction） ＜40％ by echocardiography 1 week after MI （PBS group,bFGF group,PBS＋ADSC group,ADSC＋bFGF group）.bFGF was co-injected with ADSC into the left ventricular wall of MI rats.After 4 weeks,MI size,left ventricular wall thickness,MI parts micro-vessel density,ADSC differentiation at the MI site were detected and cardiac function measured by echocardiography.Immunohistochemistry and immunofluorescence were used to evaluate possible mechanism.Results ADSC were successfully isolated and most ADSC showed positive and CD90 and CD29 and negative,CD45 and CD34 by flow cytometry.ADSC could be differentiated into adipogenic,and osteogenic cells.PBS ＋ ADSC group and ADSC ＋ bFGF group showed statistically significant differences of angiogenesis,ventricular wall thickness MI size,MI regional capacity compared to PBS groups,especially ADSC＋bFGF group （P＜0.01）.After 4 weeks of ADSC transplantation into rats,the cells could differentiate into positive cTnT,SMA positive cells and vWAg positive cells.Cardiac function test results showed that PBS＋ADSC group had the same ability of repairment as ADSC＋bFGF group.Conclusion Treatment of cotransplanting ADSC with bFGF can promote the regeneration of infarcted myocardium and blood vessels,and improve the cardiac function.