Micro-dsytrophin基因修饰的骨髓间充质干细胞移植后mdx鼠膈肌病理改变

目的探讨自体骨髓间充质干细胞(mMSCs)携带外源micro-dystrophin基因移植治疗mdx鼠对膈肌的影响。方法采用脂质体介导micro-dystrophin基因转染mdx小鼠mMSCs(microdys-mMSCs),通过尾静脉注射移植治疗mdx鼠,对照组移植等量的PBS液。在移植后4周、8周、12周和16周测定受体鼠血清肌酸激酶(CK)值、对膈肌进行HE染色、计数核中心移位纤维(CNF)比率,并用免疫荧光及RT-PCR法检测micro-dystrophin的表达。结果移植后血清CK值持续下降,各时间点膈肌病理改变较对照组有所改善,CNF比率下降,差异有统计学意义。免疫荧光可以检测到部分肌细胞膜能表达micro-dystrophin蛋白,并随移植时间延长micro-dystrophin阳性肌纤维比率增加,RT-PCR检测micro-dystrophin的mRNA有相同的趋势。结论 microdys-mMSCs移植治疗mdx鼠可以在受体鼠膈肌检测到micro-dystrophin的表达,减轻肌肉损伤并且改善mdx鼠膈肌的病理改变。

Pathological change of diaphragm in mdx mice after transplantation of bone marrow mesenchymal stem cells modified by micro - dys- trophin gene

Objective To study the change of diaphragm in mdx mice after transplantation of bone marrow mesenchymal stem cells modi- fied by micro - dystrophin gent. Methods The plasmid peDNA3.1 （ ＋ ）/micro - dystrophin was transfeeted into mMSCs by lipofeetamine. Then the micmdys - mMSCs were transplanted into mdx through the tail vein. PBS was injeeted into control group through the same way. The CK of miee from different time points was detected. The diaphragms from miee at each time point were sectioned. After HE staining, the rate of centrally nu- cleated fibers （CNF） was calculated. Expression of micro - dystrephin was detected using the method of and RT - PCR. Re- suits After transplantation of microdys - mMSCs, the level of CK continued to drop and the pathological change in diaphragm improved. Com- pared with non - transplanted mice, the CNF rate in the diaphragm of mice was reduced statistically. Over time, the dystrophin positive fibers con- tinued to increase in the diaphragm. RT - PCR showed the same trend in these samples. Conclusion Micro - dystrophin positive ceils could be tested in the diaphragm of mdx mice after transplantation of genetically modified MSCs by micro - dystrophin gene. The muscle injury reduced and the pathological change improved in transplanted mdx mice.