携人胰岛素样生长因子-1基因的成肌细胞移植后在体存活及产物表达的研究

Survival and hIGF-1 Expression of Transplanted Myoblast Carrying hIGF-1 in vivo

目的：观察并探讨携人胰岛素样生长因子-1（hIGF-1）基因的成肌细胞移植入雄性C3H小鼠体内后的存活、转归以及移植后hIGF-1基因的表达.方法：84只雄性C3H小鼠随机（20～30g,7～11w）分为A组（正常小鼠转基因细胞移植组）、B组（正常小鼠空白成肌细胞移植组）、C组（受伤小鼠转基因细胞移植组）和D组（受伤小鼠空白成肌细胞移植组）,每组20只,另4只作正常对照.A、B两组分别于右侧腓肠肌中段注射转基因成肌细胞或空白成肌细胞;C、D两组以重力打击造成小鼠右侧腓肠肌中段钝挫伤,伤后第3天分别于致伤局部注射转基因成肌细胞或空白成肌细胞.注射细胞后第2、5、10、20、30天,各组随机抽取4只小鼠处死,取右侧腓肠肌中段检测,Br-dU免疫组化染色检测外源细胞在体内的存活情况;4组另行hIGF-1免疫组化染色及实时PCR检测外源转基因细胞在体内表达hIGF-1情况.结果：各组小鼠均有BrdU免疫组化阳性染色,A、C两组均有hIGF-1 mRNA表达及hIGF-1分泌,B、D两组未检测到hIGF-1 mRNA表达及hIGF-1分泌.结论：携hIGF-1基因的成肌细胞移植入正常及钝挫伤小鼠体内后,可存活一段时间,并能稳定地分泌hIGF-1因子.

Objective To observe the survival of myoblasts carrying hIGF- 1 gene transplanted into the mice, and the expression of hIGF - 1 in the transplanted mice. Methods Eighty four male C3H mice（20- 30g,7 -11w）were divided into four groups： group A, B, C and D （20 mice each group）, and the remaining four mice were used as normal control. At the middle of the right gastrocnemius muscle, the mice in group A and B were injected with 1× 10^6 myoblasts either carried with or without hIGF- 1 gene. Muscle contusion at the middle of the right gastrocnemius muscle of the mice in group C and D was produced. At day 3 following injury, they were injected with 1×10^6 myoblasts either carried with hIGF - 1 gene （group C） or without hIGF - 1 gene （group D）. At the day 2, 5, 10, 20, 30 after injection, four mice of each group were sacrificed randomly. BrdU staining in all mice were performed to evaluate cells surviving, and the expression level of hIGF - 1 in group A and C was assessed by immunohistochemical staining and real - time - PCR. Results The BrdU staining in both normal and injured mice transplanted with myoblast carried with or without hIGF - 1 were positive, hIGF- 1 was expressed and secreted in both group A and C. Conclusion The myoblast carrying hIGF- 1 gene transplanted into normal or injured mice can survive for a certain period of time, and can secrete hIGF- 1.