携带Ad.ANG的骨髓基质细胞移植于缺血心肌的实验研究  

Studies on transplantation of bone marrow stromal cells transferred by angiogenin gene in ischemic myocardium

作　　者：李志刚[1] 梅举[1] 张宝仁[1] 

机构地区：[1]第二军医大学长海医院胸心外科,上海200433

出　　处：《中华胸心血管外科杂志》2003年第1期43-46,共4页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：国家自然科学基金资助项目 ( 30 170 935 )

摘　　要：目的　利用编码血管生长素基因的腺病毒 (Ad .ANG)转染骨髓基质细胞 (BMSCs) ,并移植于缺血心肌 ,研究其对心功能的保护作用。方法　Ad .ANG体外转染BMSCs后 ,通过ELISA方法检测ANG的表达和分泌情况 ;将转染Ad .ANG的BMSCs移植于同基因背景Lewis大鼠梗死心肌 ,通过EF、左室舒张末期容积等超声指标研究其对心功能的保护作用 ,另外通过免疫组化、电镜研究细胞存活、分化和血管新生情况。结果　Ad .ANG转染BMSCs后 ,在培养上清中有大量ANG出现 ,4～ 7d时表达达到高峰 ,15d后仍可检测到蛋白表达。移植于缺血心肌 4周后 ,实验组在心功能改善方面明显好于单纯细胞移植组 (P <0 0 5 )、Ad .ANG治疗组 (P <0 0 1)和空白对照组 (P <0 0 1) ,在促血管新生方面 ,实验组也明显优于以上 3组。结论　转染Ad .ANG后的BMSCs可获得很高的表达水平 ,移植于缺血心肌后 ,在改善心功能和促进血管新生方面明显优于单纯细胞移植组和基因治疗组。Objective: To evaluate the protective effect of bone marrow stromal cells (BMSCs) transferred by Ad·ANG ex vivo on ischemic myocardium. Methods: ELISA method was used to assay the expression and secretion of angiogenin (ANG) after Ad·ANG transfection of BMSCs ex vivo. Then BMSCs with Ad·ANG were transplanted into ischemic myocardium of isogenic Lewis rats. 4 weeks later, the parameters of heart function, such as EF and EDLV, were examined by echocardiography. Survival and differentiation of transplanted BMSCs and angiogenesis were appraised by histology and transmission electron micrography. Results: ANG was found in both lysate and culture medium after transfection of BMSCs. A maximum expression of ANG was observed at 4-7 days after transfection and could still be assayed 15 days later. 4 weeks later after transplantation in the BMSCs with Ad·ANG group, heart function improved better than the single BMSCs group(P<0.05), Ad·ANG group (P<0.01) and blank control group (P<0.01). Angiogenesis in test group was more obvious than three control groups(P<0.01). Conclusion: Satisfactory expression of ANG can be obtained Ad·ANG was transferred into BMSCs ex vivo. After transplantation of BMSCs with Ad·ANG into ischemic myocardium, improvement of heart function and angiogenesis are more obvious than single BMSCs transplantation and Ad·ANG injection.

关 键 词：心肌缺血 骨髓细胞 新生血管化 生理性 腺病毒 移植 

分 类 号：R457.7[医药卫生—治疗学]