成肌分化因子D高表达肌卫星细胞改善大鼠失神经肌肉萎缩的研究  

The effect of high expression of myogenic differentiation factor D in muscle satellite cells on prevention of denervation muscle atrophy： an experimental study in rats

作　　者：石海飞[1] 沈辉[1] 吴守成[1] 卢荟[1] 顾玉东[2] 

机构地区：[1]浙江大学医学院附属第一医院手外科,杭州310003 [2]复旦大学附属华山医院,上海

出　　处：《中华手外科杂志》2014年第5期387-390,共4页Chinese Journal of Hand Surgery

基　　金：卫生部手功能重建重点实验室开放课题;浙江省自然科学基金（LQ12H06004）;浙江省医药卫生科学研究基金（2009A068）;浙江省中医药科学研究基金计划（2010ZB064）

摘　　要：目的 研究转染了成肌分化因子D （MyoD）高表达质粒的肌卫星细胞（Muscle satellite cell,MSC）移植对改善大鼠腓肠肌失神经萎缩的作用.方法 构建MyoD高表达质粒转染大鼠MSC.将高表达MyoD的MSC移植到大鼠坐骨神经离断模型.术后2、4周取材进行肌湿重维持率检测,HE染色后检测肌纤维横截面积以及Real-time PCR（RT-PCR）检测MyoD基因表达.结果 转染后激光共聚焦显微镜检测可见MSC表达MyoD基因,转染率为43％.MyoD高表达MSC注射组腓肠肌肌湿重维持率高于其他两组,差异有统计学意义（P＜0.05）.RT-PCR检测提示MyoD高表达MSC注射组MyoD基因表达较其他两组更高,差异有统计学意义（P＜0.05）.2周及4周空白对照组出现细胞变性坏死,胞质内有空泡形成,肌纤维断裂.MSC注射组肌组织变性坏死后可见少量肌细胞修复.MyoD高表达MSC注射组可见变性坏死肌组织有肌细胞修复.2周及4周MyoD高表达MSC注射组肌细胞横截面积高于其他两组,差异有统计学意义（P＜0.05）.结论 MyoD高表达质粒的MSC移植有改善大鼠腓肠肌失神经萎缩的作用,其分子机制有待进一步研究.Objeetive To study the effect of implantation of muscle satellite cells （MSCs） transtected with myogenic differentiation factor D （MyoD） high expression plasmids on denervated gastrocnemius muscle atrophy in rats.Methods Plasmid of high expression of MyoD was constructed and used to transfect MSCs derived from Sprague-Dawley rats.These transfected MSCs were transplanted into the denervated gastrocnemius muscles in a rat sciatic nerve transection model.At 2 and 4 weeks postoperatively,gastrocnemius muscles were harvested for measurement of wet muscle weight and muscle fiber cross sectional area from H＆E stained tissue sections.The expression of MyoD was quantified with real-time PCR.Results Confocal microscopy detected MyoD expression in the transfected MSCs with a transfection efficiency of 43 %.Gastrocnemius muscle wet weight of the MyoD transfected MSC group was higher than those of the other groups （P 〈 0.05）.The level of MyoD gene expression was higher in the MyoD transfected MSC group than in the other groups （P 〈 0.05）.Muscle cell degeneration and cell death was noted in the control group at 2 and 4 weeks postoperatively.Vacuole formation and myofiber rupture were also observed.Myofiber regeneration was present in the MSC group and MyoD transfected MSC group.Muscle fiber cross sectional area in the MyoD transfected MSC group was higher than the other groups at both 2 and 4 week time points.The differences were statistically significant （P 〈 0.05）.Conclusion Transplantation of high MyoD expression MSCs can delay denervation atrophy of gastrocnemius muscle in rats.

关 键 词：肌萎缩 骨骼肌卫星细胞 成肌分化因子D 失神经 基因治疗 

分 类 号：R746.4[医药卫生—神经病学与精神病学]