EGCG对乐果致小鼠骨骼肌内质网应激及成肌分化受损的保护作用  

Protective effect of EGCG on endoplasmic reticulum stress and impaired myogenic differentiation induced by dimethoate in mouse skeletal muscle

作　　者：康国静 陈嘉伟 邹辉[1,2,3] 顾建红 袁燕[1,2,3] 卞建春 刘学忠[1,2,3] KANG Guo-jing;CHEN Jia-wei;ZOU Hui;GU Jian-hong;YUAN Yan;BIAN Jian-chun;LIU Xue-zhong(College of Veterinary Medicine,Yangzhou University;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses;Joint International Research Laboratory of Agriculture and Agri-Products Safety,The Ministry of Education of China,Yangzhou University)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [3]扬州大学教育部农业与农产品安全国际合作联合实验室,江苏扬州225009

出　　处：《中国兽医科学》2023年第10期1333-1340,共8页Chinese Veterinary Science

基　　金：江苏省农业科技自主创新资金项目[CX(18)3022];江苏省高校优势学科建设工程资助项目(PAPD)。

摘　　要：为探究表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)在乐果(dimethoate,DIM)经皮肤慢性暴露致小鼠骨骼肌损伤过程中是否具有保护作用,将6周龄雄性ICR小鼠随机分为对照组(腹腔注射0.2 m L生理盐水、背部涂抹0.1 m L生理盐水)、EGCG组(腹腔注射0.2 m L EGCG、背部涂抹0.1 m L生理盐水)、DIM组(腹腔注射0.2 m L生理盐水、背部涂抹0.1 m L DIM)、DIM+EGCG组(腹腔注射0.2 m L EGCG、背部涂抹0.1 m L DIM),记录小鼠体重变化,检测小鼠骨骼肌MDA含量及CAT、T-SOD、T-AOC的活性,利用全自动生化分析仪检测血清CK和LDH活性,利用透射电镜观察骨骼肌超微结构,利用Western-blot检测Bip、PERK/eIF2α通路相关蛋白及成肌分化相关蛋白表达水平。结果显示,与对照组相比,DIM组第2~3周小鼠体重降低显著(P<0.05);第4~8周小鼠体重降低极显著(P<0.01);MDA含量升高极显著(P<0.01),CAT、T-SOD、T-AOC活性降低极显著(P<0.01);血清CK、LDH活性升高极显著(P<0.01);超微结构损伤;内质网应激标志蛋白Bip、PERK/eIF2α通路磷酸化蛋白表达水平显著升高(P<0.05);成肌分化关键蛋白的表达水平降低极显著(P<0.01)。与DIM组相比,DIM+EGCG组小鼠体重无显著差异,总体呈缓慢上升趋势;MDA含量降低极显著(P<0.01),CAT、T-SOD、T-AOC活性升高显著(P<0.05)或升高极显著(P<0.01);血清CK、LDH活性降低显著(P<0.05)或降低极显著(P<0.01);骨骼肌超微结构损伤有所缓解;Bip及PERK/eIF2α通路磷酸化蛋白表达水平降低显著(P<0.05)或降低极显著(P<0.01);成肌分化关键蛋白的表达水平升高显著(P<0.05)或升高极显著(P<0.01)。结果表明,DIM暴露导致了小鼠骨骼肌损伤,引起内质网应激并激活了下游的PERK/eIF2α通路,使小鼠骨骼肌成肌分化受损;EGCG能缓解DIM所致的氧化损伤,提高抗氧化能力,降低血清CK、LDH的活性,显著抑制内质网应激及下游PERK/eIF2α通路磷酸化水平,促进成肌分化蛋白的表达,改To investigate whether epigallocatechin gallate(EGCG)has a protective effect on skeletal muscle injury induced by chronic exposure of dimethoate(DIM)through the skin,the 6-week-old male ICR mice were randomly divided into control group(0.2 m L intraperitoneal injection of normal saline and 0.1 m L on the back),EGCG group(0.2 m L intraperitoneal injection of EGCG and 0.1 m L on the back),DIM group(0.2 m L intraperitoneal injection of normal saline and 0.1 m L DIM on the back),DIM+EGCG group(0.2 m L intraperitoneal injection of EGCG and 0.1 m L DIM on the back).The changes of body weight were recorded,and MDA content and the activities of CAT,T-SOD and T-AOC in skeletal muscle were detected,CK and LDH activities in serum were detected by automatic biochemical analyzer,and ultrastructure of skeletal muscle was observed by transmission electron microscope.The expression levels of Bip,PERK/e IF2αpathway related proteins and myogenic differentiation related proteins were detected by Western-blot.The results showed that compared with the control group,the body weight of DIM group was significantly decreased at 2 to 3 weeks(P<0.05);the body weight of mice was extremely significantly decreased from 4 to 8 weeks(P<0.01).The content of MDA in DIM group was extremely significantly increased(P<0.01),and the activities of CAT,T-SOD and T-AOC were extremely significantly decreased(P<0.01).Serum CK and LDH activities were extremely significantly increased(P<0.01).Ultrastructural damage.The expression levels of endoplasmic reticulum stress markers Bip and PERK/e IF2αpathway phosphorylated proteins were significantly increased(P<0.05).The expression level of key protein of myogenic differentiation was extremely significantly decreased(P<0.01).Compared with DIM group,there was no significant difference in body weight of DIM+EGCG group,and the overall body weight showed a slow upward trend.MDA content of DIM+EGCG group was extremely significantly decreased(P<0.01),and activities of CAT,T-SOD and T-AOC were significantly increased(

关 键 词：乐果 EGCG 骨骼肌 内质网应激 小鼠 

分 类 号：S852.35[农业科学—基础兽医学]