兔骨髓间充质干细胞mHCN4基因修饰后异体心外膜下移植区细胞形态学动态演变  

Dynamic morphology succession of mHCN4 gene modified rabbit MSCs after allogeneic subepicardial transplantation

作　　者：张志辉[1] 程军[1] 宋治远[1] 

机构地区：[1]第三军医大学西南医院心内科重庆市介入心脏病学研究所,重庆400038

出　　处：《中国心脏起搏与心电生理杂志》2009年第6期524-527,共4页Chinese Journal of Cardiac Pacing and Electrophysiology

基　　金：国家自然科学基金(项目编号:30871060);重庆市自然科学基金重点项目(项目编号:2008BA5005)资助

摘　　要：目的观察兔骨髓间充质干细胞(MSCs)mHCN4基因修饰后异体心外膜下移植区细胞形态学的演变情况,探讨细胞移植治疗缓慢性心律失常的可行性。方法日本大耳白兔50只随机分为对照组与细胞移植3天、1周、2周、4周组(每组各10只)。开胸直视下将0.2 ml含1×106个/毫升转染mHCN4基因的兔MSCs悬液注入兔左室游离壁心外膜下,对照组注入等体积生理盐水。HE染色观察细胞形态、免疫荧光检测EGFP及mHCN4表达。结果共有46只兔完成实验。对照组表现为轻微炎症反应,动态演变不明显。移植组术后3天时可见大量炎性细胞浸润,移植细胞多为圆形,与周围心室肌界限明显;术后1周时移植细胞数量明显减少,可见部分细胞呈短梭形;术后2周时炎症细胞及移植细胞进一步减少,梭形移植细胞数量增多;术后4周时存活移植细胞形态基本呈长梭形,与周围心室肌界限较模糊。免疫荧光于术后3天时即可检出EGFP和mHCN4阳性表达;术后2周时移植细胞形态已呈典型长梭形,与正常心室肌细胞相邻生长。结论mHCN4基因修饰的兔MSCs心外膜下移植后存活细胞从圆形逐渐伸展为梭形,可成功表达EGFP和mHCN4通道蛋白,到术后2周时形态已接近于心室肌细胞。Objective To explore the feasibility of cell transplantation in treatment of bradyarrythmias through observation on Dynamic morphology succession of mHCN4 gene modified rabbit MSCs after allogeneic subepicardial transplantation. Method Fifty rabbits were randomly divided into 5 groups （ 10 in each group） ： control, 3d ,1w,2w and 4w groups after cell transplantation. 0.2 ml suspension including 1 × 10^6/ml mHCN4 gene modified rabbit MSCs was injected into the subepicardial free wall of left ventricle while normal sodium was injected in the control group. HE staining was used for observation of cell morphology and immunofluorescence for EGFP and mHCN4 detection. Results Forty-six rabbits were used in the experiment. Control group showed slight inflammatory reaction with obsolete dynamic variation. A great quantity of inflammatory cells infiltrated at 3 days after transplantation, and most transplanted cells were round, with clear bouncary from normal surrounding myocardium. In lw after operation group transplanted cell decreased significantly and some of them showed short fusiform shape. Inflammatory cells and transplanted cells obviously decreased and fusiform shape cells increased at 2 weeks after transplantation. In 4w after cell transplantation group most survival transplanted cells were long fusiform shape and with undefined bouncary from normal myocardimn. EGFP and mHCN4 positive expression were detected by immnnofluorescence at 3 days after transplantation. In 2w after transplantation, the transplanted cells were typical long fusiform shape and located with surrounding normal myoccardium cells. Conclusion Morphology of survival transplanted mHCN4 gene modified rabbit MSCs becomes fusiform shape from round shape gradually and can express EGFP and mHCN4 protein. The shape of transplanted cells is like ventricular myocardium ceils at 2 weeks after transplantation.

关 键 词：心血管病学 mHCN4 骨髓间充质干细胞 移植 生物起搏 

分 类 号：R318.11[医药卫生—生物医学工程]