Zhang 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-lin
Department of Neurology, First Hospital of Sun Yat-sen University, Guangzhou 510080, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2006 Jan;26(1):53-8.
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).
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.
The proportion of centrally nucleated fibers (CNF) in the diaphragm muscle of the recipient mdx mice was (15.58+/-0.91) %, (12.50+/-1.87) % and (10.17+/-1.17) % at 4, 8 and 12 weeks after BMT, respectively, significantly smaller than that of untreated mdx mice [(19.5+/-1.87) %], and the fibers after BMT showed less inflammatory infiltration. Compared with the untreated mice, the recipient mdx mice showed green fluorescence on significantly more diaphragm muscle cell membranes [with the proportion of dystrophin-positive fibers of (1.00+/-0.32) %, (6.00+/-1.05) % and (11.92+/-1.11) % at 4, 8, and 12 weeks after BMT]. RT-PCR of dystrophin mRNA also demonstrated significantly higher relative levels of dystrophin in the recipient mdx mice (0.19+/-0.05, 0.26+/-0.06 and 0.36+/-0.04 at 4, 8 and 12 weeks after BMT) than in untreated mdx mice, and Sry gene was present in the recipient mice.
BMT can partially restore dystrophin expression and ameliorate the pathology in the diaphragm muscles of mdx mice, and has great potential to produce general therapeutic effect in patients with DMD.
研究骨髓干细胞移植（BMT）对杜氏肌营养不良症（DMD）小鼠模型mdx小鼠膈肌的影响。
将雄性SD大鼠的骨髓源性干细胞经尾静脉移植到18只8周龄的雌性mdx小鼠体内，分别在BMT后4周、8周和12周处死小鼠（每个时间点6只）。对小鼠的膈肌进行苏木精-伊红（HE）染色、抗肌萎缩蛋白免疫荧光检测、抗肌萎缩蛋白mRNA转录本的逆转录（RT）-PCR分析以及Y染色体性别决定区（Sry）基因的PCR分析，以年龄匹配的雌性C57小鼠和未处理的mdx小鼠作为对照。
受体mdx小鼠膈肌中中央核纤维（CNF）的比例在BMT后4周、8周和12周分别为（15.58±0.91）%、（12.50±1.87）%和（10.17±1.17）%，显著低于未处理的mdx小鼠[（19.5±1.87）%]，且BMT后的纤维炎症浸润较少。与未处理的小鼠相比，受体mdx小鼠膈肌细胞膜上出现绿色荧光的比例显著更高[BMT后4周、8周和12周抗肌萎缩蛋白阳性纤维的比例分别为（1.00±0.32）%、（6.00±1.05）%和（11.92±1.11）%]。抗肌萎缩蛋白mRNA的RT-PCR也显示，受体mdx小鼠中抗肌萎缩蛋白的相对水平显著高于未处理的mdx小鼠（BMT后4周、8周和12周分别为0.19±0.05、0.26±0.06和0.36±0.04），且受体小鼠中存在Sry基因。
BMT可部分恢复mdx小鼠膈肌中抗肌萎缩蛋白的表达并改善其病理状况，对DMD患者具有产生总体治疗效果的巨大潜力。
