CAJ | 학술논문

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.
목적 연구전염료성기분화인자D (MyoD)고표체질립적기위성세포(Muscle satellite cell,MSC)이식대개선대서비장기실신경위축적작용.방법 구건MyoD고표체질립전염대서MSC.장고표체MyoD적MSC이식도대서좌골신경리단모형.술후2、4주취재진행기습중유지솔검측,HE염색후검측기섬유횡절면적이급Real-time PCR(RT-PCR)검측MyoD기인표체.결과 전염후격광공취초현미경검측가견MSC표체MyoD기인,전염솔위43％.MyoD고표체MSC주사조비장기기습중유지솔고우기타량조,차이유통계학의의(P＜0.05).RT-PCR검측제시MyoD고표체MSC주사조MyoD기인표체교기타량조경고,차이유통계학의의(P＜0.05).2주급4주공백대조조출현세포변성배사,포질내유공포형성,기섬유단렬.MSC주사조기조직변성배사후가견소량기세포수복.MyoD고표체MSC주사조가견변성배사기조직유기세포수복.2주급4주MyoD고표체MSC주사조기세포횡절면적고우기타량조,차이유통계학의의(P＜0.05).결론 MyoD고표체질립적MSC이식유개선대서비장기실신경위축적작용,기분자궤제유대진일보연구.
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.