摘要
本试验用不同浓度(0、0.3、0.6、0.9 mmol/L)的乐果(DIM)处理C2C12细胞24 h,采用CCK-8检测细胞活力,光镜及透射电镜观察细胞形态及超微结构,Western-blot检测Bip、PERK/e IF2α通路相关蛋白及成肌分化相关蛋白MyOD、MyOG、My HC的表达水平。结果显示,与0 mmol/L组相比,0.3 mmol/L组的细胞活力、Bip及PERK/e IF2α通路相关蛋白表达水平、成肌分化相关蛋白的蛋白表达水平均无显著差异。随DIM浓度升高,细胞活力极显著降低(P<0.01);细胞形态发生改变,死细胞增多,细胞超微结构受损明显;Bip及PERK/e IF2α通路相关蛋白表达水平显著(P<0.05)或极显著(P<0.01)升高;成肌分化相关蛋白的表达水平显著(P<0.05)或极显著(P<0.01)降低。添加PERK/e IF2α通路特异性抑制剂GSK2606414与DIM共处理,相关蛋白的表达水平被扭转。结果表明,DIM导致C2C12细胞内质网应激并激活了下游的PERK/e IF2α通路,且该通路介导了DIM致C2C12细胞成肌分化受损的过程。
C2C12 cells were exposed to dimethoate(DIM)at various concentrations(0,0.3,0.6,0.9mmol/L)for 24 h.Cell viability was determined by CCK-8,cell viability morphology and ultrastructure were observed through light microscopy and transmission electron microscopy,and the expression levels of Bip,PERK/e IF2α pathway-related proteins,My OD,My OG,and My HC were detected by Western-blot.The results indicated as follows:Compared with the 0 mmol/L group,the cell activity of the 0.3 mmol/L group was higher than that of the 0 mmol/L group.The expression levels of Bip and PERK/e IF2α pathway-related proteins.There were no significant differences in the expression levels of myodifferentiation-related proteins.The cell viability decreased significantly with the increase of DIM concentration(P<0.01).The cell morphology was altered,the number of dead cells increased,and the cell ultrastructure was evidently damaged.The expression levels of Bip and PERK/e IF2α pathway-related proteins were significantly elevated(P<0.05)or(P<0.01).The expression levels of myodifferentiation-related proteins were significantly decreased(P<0.05)or significantly declined(P<0.01).PERK/e IF2α pathway-specific inhibitor GSK2606414 was added for DIM co-treatment,and the expression level of related proteins was reversed.The results demonstrated that DIM induced endoplasmic reticulum stress in C2C12 cells and activated the downstream PERK/e IF2α pathway,which mediated the impaired myoblast differentiation of C2C12 cells induced by DIM.
作者
康国静
陈嘉伟
邹辉
顾建红
袁燕
卞建春
刘学忠
KANG Guojing;CHE Jiawei;ZOU Hui;CU Jianhong;YUAN Yan;BIAN Jianchun;LIU Xuezhong(The Animal Disease Control and Diagnosis Center of Kashi Distric,Kashi 844000,China;College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China;Joint International Research Laboratory of A griculture and A gri-Products Safety,The Ministry of Education of China,Yangzhou University,Yangzhou 225009,China)
出处
《中国兽医科学》
北大核心
2025年第1期121-128,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)。
