抗肌萎缩蛋白小基因对Duchenne肌营养不良病理改变的治疗作用

目的　探讨Duchenne肌营养不良 (DMD)的病理变化和治疗方法。方法　采用重组腺相关病毒载体 (rAAV)介导的人抗肌萎缩蛋白小基因 (SMCKA3999)观察DMD的病理改变和治疗作用。将抗肌萎缩蛋白小基因SMCKA3999克隆至rAAV并包装成rAAVSMCKA3999病毒 ,将 5× 10 9病毒颗粒分多点注射于DMD模型鼠mdx腓肠肌 ,4个月后取局部肌组织 ,分别采用免疫荧光、埃文斯氏蓝染料、电镜等方法 ,观察rAAVSMCKA3999对DMD病理改变的治疗作用。结果　rAAVSMCKA3999有效稳定表达并使肌膜缺失的抗肌萎缩蛋白恢复 ,明显改善DMD肌肉病理改变。结论　rAAVSMCKA3999能显著改善mdx小鼠骨骼肌病理变化 ,有希望成为Duchenne肌营养不良有效的生物治疗药物

Adeno-associated virus vector carrying human minidystrophin gene SMCKA3999 effectively ameliorates dystrophic pathology in mdx model mice

Objective Duchenne muscular dystrophy(DMD) is the most common and letal genetic skeletal muscle disorder, caused by recessive mutations in the dystrophin gene and no treatment is available. The present paper is aimed to study if recombinant adeno associated virus vector(rAAV) mediated dystrophin minigene SMCKA3999 could effectively ameliorates dystrophic pathology in mdx model mice. Methods We used dystrophin minigene SMCKA3999 to construct rAAVSMCKA3999. When injected into the skeletal muscle of mdx mice(DMD model), we adopted methods of immunofluorescent (IF) staining, Evans Blue and electromicroscopy to observe if rAAVSMCKA3999 could effectively ameliorates dystrophic pathology in mdx model mice. Results rAAVSMCK3999 resulted in efficient and stable expression and restoration of the missing dystrophin onto the plasma membrane. rAAVSMCKA3999 can also protect myofiber membrane integrity.For the first time we prove that rAAVSMCKA3999 can improve ultrastructure changes of DMD. Conclusion We have demonstrated the effectiveness of rAAVSMCKA3999 in correcting pathology changes of skeletal muscle,which offer a promising avenue for DMD gene therapy.