免疫组化联合放射自显影术观察骨髓间充质干细胞在肌营养不良鼠表达抗肌萎缩蛋白  

Immunohistochemistry combined with autoradiography in observation of dystrophin expression in mesenchymal stem cells of mdx mice

作　　者：李经伦[1] 吕英姿[1] 何晓英[1] 冯善伟[2] 柳太云[2] 张成[2] 

机构地区：[1]泸州医学院附属医院神经内科,四川泸州646000 [2]中山大学附属医院神经内科

出　　处：《泸州医学院学报》2013年第4期320-324,共5页Journal of Luzhou Medical College

基　　金：国家自然科学基金资助项目(30370510)

摘　　要：目的:用免疫组化联合放射自显影术观察移植骨髓间充质干细胞表达dystrophin蛋白情况。方法:用3H-TdR标记的骨髓间充质干细胞经静脉移植肌营养不良鼠(mdx),在1、2、3、4月取小鼠腓肠肌肉,先进行免疫组化染色,再将标本立即进行放射自显影,3月后再将组织HE染色,显微镜下观察干细胞是否表达了dystrophin蛋白。同时用激光共聚焦显微镜(LSCM)观察BrdU标记的间充质干细胞移植mdx鼠后的dystrophin蛋白表达为比较标准。结果:两法均观察到移植的干细胞表达了dystrophin蛋白,且观察到的阳性细胞数比较无统计学意义;干细胞表达dystrophin蛋白随着时间有逐渐增高趋势。结论:免疫组化联合放射自显影技术可以用于对干细胞移植后蛋白表达的示踪观察,干细胞移植对肌营养不良具有治疗前景。Objective： To study dystrophin expression in mesenchymal stem cells from mdx mice by immunohistochemistry combined with autoradiography. Methods： Bone marrow mesenchymal stem cells labeled with 3H-TdR were transferred intravenously into 30 irradiated mdx mice 1、2、3 and 4 m after transfer, leg muscle of mdx rats was cryosected and fixed, then stained with routine immunohistochemical method 3 months later,it was stained by hematoxylin and eosin（HE）, and observed by microscope. The other cryosections labeled by BrdU were observed by LSCM . Results： The distrophin expression of MSCs was detected by both methods without statistic difference.The expression increased gradually with the time. Conclusion： Dystrophin expression of MSCs can be observed by autoradiography combined with immunohistochemical technology. It may be applied to the evaluation of MSCs transplantation in myodystrophic patient in future.

关 键 词：免疫组化 放射自显影术 骨髓间充质干细胞 肌营养不良鼠 抗肌萎缩蛋白 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]