摘要
目的研究miRNA-206通过Akt信号通路抑制失神经肌肉萎缩的机制。方法构建小鼠失神经支配肌肉萎缩模型,荧光定量PCR检测miRNA-206变化;小鼠肌肉原位注射miRNA-206 agomir或miRNA-206 sponge,3 d后小鼠建造肌肉失神经支配模型,7 d后免疫印迹检测肌肉萎缩标志蛋白Fbx32和TRIM63的表达量;以荧光素酶报告实验验证miRNA-206和Akt 3′UTR区域结合。结果荧光定量PCR结果显示,miRNA-206在失神经导致的肌肉萎缩中下调。小鼠肌肉原位注射miRNA-206 agomir,免疫印迹实验结果显示,肌肉萎缩标志蛋白Fbx32和TRIM63表达明显降低;失神经支配小鼠肌肉原位注射miRNA-206 sponge,Fbx32和TRIM63表达明显升高。通过荧光素酶实验证明了miRNA-206和Akt 3′UTR区域结合。进一步的免疫印迹实验结果显示,抑制miRNA-206表达时,Akt表达下调;过表达miRNA-206时,Akt表达上调。结论miRNA-206通过Akt信号通路抑制失神经肌肉萎缩。
Objective To explore the mechanism of miRNA-206 inhibiting denervated muscle atrophy through Akt signaling pathway.Methods The denervated muscle atrophy model in mice was established,and the changes of miRNA-206 were detected by fluorescence quantitative PCR.The miRNA-206 agomir or miRNA-206 Sponge was injected into the muscles of mice in situ and the mice underwent denervation after 3 days.The expression of muscle atrophy marker protein Fbx32 and TRIM63 was detected by Western blot after 7 days.The binding of miRNA-206 to Akt 3′UTR was verified by luciferase reporting assay.Results The fluorescence quantitative PCR results showed that miRNA-206 was down-regulated in denervation induced muscle atrophy.The intramuscular injection of miRNA-206 agomir was performed in denervation mice and Western blot results showed that the expressions of muscle atrophy marker protein Fbx32 and TRIM63 were significantly decreased.The intramuscular injection of miRNA-206 sponge was performed in denervation mice and Western blot results showed that the expressions of Fbx32 and TRIM63 were significantly increased.The binding of miRNA-206 to Akt 3′UIR was demonstrated by luciferase reporting assay.The Western blot results showed that Akt expression was down-regulated when miRNA-206 expression was inhibited and Akt expression was up-regulated when miRNA-206 was overexpressed.Conclusion The miRNA-206 inhibits denervated muscle atrophy through Akt signaling pathway.
作者
杨卫可
贾瑞平
田致忠
杨卫强
郭新军
郭继强
Yang Weike;Jia Ruiping;Tian Zhizhong;Yang Weiqiang;Guo Xinjun;Guo Jiqiang(Department of Orthopedics,Xinxiang Central Hospital,Henan 453000,China;Department of Immunology,School of Basic Medical Science,Xinxiang Medical College,Henan 453003,China)
出处
《中华手外科杂志》
CSCD
北大核心
2021年第5期377-381,共5页
Chinese Journal of Hand Surgery
基金
河南省卫生健康委员会基金(LHGJ20191326)。