摘要目的 探讨失神经大鼠腓肠肌细胞中叉头蛋白转录因子3a(fork-head protein,FoxO3a)及MuRF-1的表达规律.方法 72只Wistar大鼠随机分为12组,每组6只,切断大鼠右侧坐骨神经,左侧行假手术,分别于术后0、1、2、3、4、5、6、7、10、14、21、28d各处死一组大鼠,取右侧腓肠肌,于-70℃冰箱保存.分别使用实时荧光定量PCR法和Western Blot法对其中的FoxO3a和MuRF-1的基因和蛋白进行检测.结果 MuRF-1的基因表达在失神经第4天达到峰值(18.12倍),失神经14d后下降至正常.FoxO3a的基因表达在失神经第6天达到峰值(7.06倍),而后虽缓慢下降,但仍高表达.FoxO3a和MuRF-1的蛋白含量随失神经时间的延长逐渐升高.结论 FoxO3a和MuRF-1的表达和蛋白改变与失神经骨骼肌萎缩关系密切,可以作为药物治疗和基因治疗的靶点之一.

abstractsObjective To investigate the expression patterns of FoxO3a and MuRF-1 in the myocytes of denervated gastrocnemius muscle in rats. Methods Seventy-two Wistar rats were randomly divided into 12 groups with 6 rats each.The right sciatic nerve was transected while the left underwent a sham operation.One group of rats was sacrificed at postoperative day0,1,2,3,4,5,6,7,10,14,21 and 28 respectively to harvest the right gastrocnemius muscles.The samples were frozen at - 70℃.Real-time PCR was used to detect the mRNA level of FoxO3a and MuRF-1,and western blot was done to detect the protein concentration of FoxO3a and MuRF-1 for each sample. Results The mRNA level of MuRF-1 reached its peak at 4 days after the denervation (18.12 folds),and returned to normal level after 14 days.The mRNA level of FoxO3a peaked at 6 days after the denervation (7.06 folds),while maintaining at a high level despite the slow decline.The protein concentration of FoxO3a and MuRF-1 increased with the days of denervation. Conclusion The expressions of FoxO3a and MuRF-1 are closely related to the process of muscle atrophy after denervation.Both FoxO3a and MuRF-1 can be the targets for drug treatment or gene therapy to delay denervation atrophy.