Nrf2蛋白在骨骼肌损伤修复中的时序性表达及作用  

Time-dependent Expression of Nrf2 Protein and its Effects on Wound Repair in Skeletal Muscle

作　　者：张小红[1,2] 刘季 李娜[1] 杜秋香[1] 王英元[1] 孙俊红[1] ZHANG Xiao-hong;LIU Ji;LI Na;DU Qiu-xiang;WANG Ying-yuan;SUN Jun-hong(School of Forensic Medicine, Shanxi Medical University, Taiyuan 030001, China;Zouping Public Security Bureau, Zouping 256200, China;Traffic Police Detachment of Guangzhou Municipal Public Security Bureau, Guangzhou 510000, China)

机构地区：[1]山西医科大学法医学院,山西太原030001 [2]邹平县公安局,山东邹平256200 [3]广州市公安局交通警察支队,广东广州510000

出　　处：《法医学杂志》2018年第1期18-22,共5页Journal of Forensic Medicine

基　　金：国家自然科学基金面上资助项目(81571852);山西省回国留学人员科研资助项目(2015-052)

摘　　要：目的通过检测骨骼肌损伤后核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)蛋白的表达变化,探讨其在骨骼肌损伤修复过程中的表达变化规律及作用。方法取雄性SD大鼠腓肠肌内注射心脏毒素,制作骨骼肌损伤模型,分别取正常对照组和损伤后1h、4h、8h、12h、16h、1d、3d、5d、7d、9d、13d、17d、21d的损伤处组织,采用HE染色法观察骨骼肌损伤修复过程中的形态学变化,检测活性氧(reactive oxygen species,ROS)相对含量,Western印迹法及免疫荧光染色检测Nrf2蛋白表达水平。结果大鼠骨骼肌损伤后,损伤区依次出现以中性粒细胞、单核细胞为主的炎症细胞浸润,损伤后5d大量新生肌细胞生成,至21d基本完成肌细胞修复。与正常对照组相比,大鼠骨骼肌损伤后:ROS相对含量仅第12h降低(P<0.05);Nrf2蛋白16h表达升高(P<0.05);Nrf2阳性核率在损伤后增多,16h^1d达到高峰,随后逐步减少,最后恢复至对照水平(P<0.05)。结论 Nrf2蛋白参与了骨骼肌损伤修复的调控,Nrf2阳性核率随损伤时间呈时序性变化,可用于骨骼肌损伤时间的推断。Objective To detect the expression of nuclear factor erythroid2-related factor2(Nrf2)to explore its change rule and effects in the repair process of skeletal muscle.Methods An injury model of skeletal muscle was established by injection of cardiotoxin into the gastrocnemius muscle of SD male rats.The muscle samples were taken from control group and injury group at each time point(1h,4h,8h,12h,16h,1d,3d,5d,7d,9d,13d,17d,21d).The morphological changes of skeletal muscle in repair process were observed by H&E staining,and the content of reactive oxygen species(ROS)was detected.The expression level of Nrf2protein was investigated by Western blotting and immunohistochemistry staining.Results After the skeletal muscle injury,the injury region was infiltrated with inflammatory cells which constituted mainly by lymphocyte neutrophils and mononuclear cells.There were large amounts of immature muscle cells appeared at the5th day and the muscle repair was completed at the21th day.Comparing with control group,the ROS content decreased in injury group at the12th hour(P<0.05),while Nrf2expression increased at the16th hour(P<0.05).Moreover,the positive rate of Nrf2increased after injury,and peaked at16h-1d,then decreased gradually and returned to the level of control group(P<0.05).Conclusion The Nrf2protein involves in the regulation of the repair of skeletal muscle injury.The positive rate of Nrf2shows a time-dependent expression,which can be used to estimate the wound age of skeletal muscle.

关 键 词：法医病理学 肌 骨骼 活性氧 核因子E2相关因子2 氧化性应激 损伤时间推断 

分 类 号：DF795.1[医药卫生—法医学]