异体骨髓干细胞移植后假肥大型肌营养不良症模型鼠膈肌dystrophin.docVIP

异体骨髓干细胞移植后假肥大型肌营养不良症模型鼠膈肌dystrophin表达及病理改变 作者：张雅妮；张成；于美娟；王淑辉；李美山；黄慧；熊符；冯善伟；柳太云；卢锡林　　(中山大学第一医院神经科，广东??广州??510080) 摘要：目的??探讨骨髓干细胞移植对假肥大型肌营养不良症(DMD)模型鼠-mdx鼠膈肌的治疗效果。方法??取雄性SD大鼠骨髓干细胞经尾静脉植入放疗处理后的8周龄雌性mdx鼠(n=18)，于移植后4、8、12周各取6只mdx鼠的膈肌行HE染色、抗肌萎缩蛋白(dystrophin)免疫荧光检测以及dystrophin mRNA的RT-PCR分析，同时用正常C57鼠及放疗而未移植的mdx鼠作为对照，另进行PCR反应检测实验鼠膈肌内Sry(Y染色体的性别决定区)基因。结果 移植后mdx鼠膈肌间质内炎性细胞浸润较放疗而未移植mdx鼠有所减少；移植后核中心移位纤维比例[移植后4、8、12周分别为(15.58±0.91)%、(12.50±1.87)%、(10.17%±1.17)%]较未移植mdx鼠(19.5±1.87)%显著减少。移植后dystrophin免疫荧光阳性细胞比例[移植后4、8、12周分别为(1.00±0.32)%、(6.00±1.05)%、(11.92±1.11%)]较未移植mdx鼠(0.17±0.41)%显著增加。RT-PCR结果显示C57鼠膈肌的dystrophin mRNA相对含量(0.63±0.04)最高；而未移植mdx鼠的膈肌中未检测到dystrophin mRNA；移植后的mdx鼠膈肌中mRNA的表达水平(移植后4、8、12周分别为0.19±0.05、0.26±0.06、0.36±0.04)随时间推移逐渐增高；移植后各时间点mdx鼠膈肌Sry基因均为阳性。结论??骨髓干细胞系统移植mdx鼠可以恢复部分膈肌的dystrophin表达，改善膈肌的病理,骨髓干细胞移植有希望成为全身治疗DMD的有效方法。 关键词：假肥大型肌营养不良症；mdx鼠；膈肌；骨髓移植；抗肌萎缩蛋白 中图分类号：R392.4; R457.7??文献标识码：A??文章编号：1673-4254(2006)01-0053-06 Dystrophin expression and pathology of diaphragm muscles of mdx mice after xenogenic bone marrow stem cell transplantationZHANG Ya-ni；ZHANG Cheng；YU Mei-juan；WANG Shu-hui；LI Mei-shan；HUANG Hui；XIONG Fu；FENG Shan-wei；LIU Tai-yun；LU Xi-linDepartment of Neurology, First Hospital of Sun Yat-sen University, Guangzhou 510080, ChinaAbstract: Objective To investigate the effect of bone marrow stem cell transplantation (BMT) on the diaphragm muscles of mdx mice, a mouse model of Duchenne muscular dystrophy (DMD). Methods The bone marrow-derived stem cells form male SD rats was transplanted through the tail vein into 18 female 8-week-old mdx mice, which were sacrificed at 4, 8 and 12 weeks after BMT (6 at each time point), respectively. The diaphragm muscles of the mice were subjected to HE staining, immunofluorescence detection of dystrophin, reverse transcription (RT)-PCR analysis of dystrophin mRNA transcripts and PCR analysis of Sry (sex-determining region on the Y chromosome) gene, with age-matched female C57 mice and untreated mdx mice as the controls. R