钼对小鼠骨骼肌上皮细胞氧化损伤及细胞骨架的影响  

Effects of Molybdenum on Oxidative Damage and Cytoskeleton in Skeletal Muscle Epithelial Cells of Mouse

作　　者：马淑浩 杨自军[1] 杜伯强[1] 李光照[1] 张才[1] 汪纪仓[1] 王宏伟[1] 孔涛[1] MA Shuhao;YANG Zijun;DU Boqiang;LI Guangzhao;ZHANG Cai;WANG Jicang;WANG Hongwei;KONG Tao(College of Animal Science and Technology,Henan University of Science and Technology,Henan Provincial Open Laboratory of Key Disciplines for Environmental and Animal Products Safety,Luoyang 471000,China)

机构地区：[1]河南科技大学动物科技学院环境与畜产品安全河南重点学科开放实验室

出　　处：《动物营养学报》2019年第12期5664-5671,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31040081)

摘　　要：本试验旨在探究钼(Mo)对小鼠骨骼肌上皮细胞氧化损伤及细胞骨架的影响。试验以小鼠原代骨骼肌上皮细胞为材料,利用Mo浓度分别为0(Ⅰ组)、0.1(Ⅱ组)、0.2(Ⅲ组)、0.4(Ⅳ组)、0.8 mmol/L(Ⅴ组)的培养基对其进行细胞培养24 h。倒置显微镜观察细胞形态变化,四甲基偶氮唑盐(MTT)法测定细胞活力,比色法检测细胞抗氧化能力及氧化损伤,实时荧光定量PCR(qRT-PCR)和蛋白质免疫印迹(Western-blot)测定细胞骨架蛋白波形蛋白(Vimentin)、α-肌动蛋白(α-actin)、β-肌动蛋白(β-actin)的mRNA和蛋白表达。结果显示:1)Ⅰ和Ⅱ组细胞形态正常,Ⅲ、Ⅳ和Ⅴ组细胞间隙增大,细胞肿大,且Ⅴ组细胞出现明显畸形。2)与Ⅰ组相比,Ⅱ、Ⅲ、Ⅳ和Ⅴ组的小鼠骨骼肌上皮细胞活力极显著下降(P<0.01)。3)与Ⅰ组相比,Ⅱ、Ⅲ、Ⅳ和Ⅴ组小鼠骨骼肌上皮细胞的超氧化物歧化酶活性和总抗氧化能力均极显著下降(P<0.01),丙二醛含量极显著升高(P<0.01)。4)与Ⅰ组相比,Ⅱ和Ⅲ组小鼠骨骼肌上皮细胞中Vimentin、α-actin和β-actin的mRNA相对表达量显著升高(P<0.05),Ⅴ组显著降低(P<0.05);Ⅲ、Ⅳ和Ⅴ组小鼠骨骼肌上皮细胞中Vimentin、α-actin和β-actin的蛋白相对表达量均显著降低(P<0.05)。综上所述,小鼠骨骼肌上皮细胞在经过高浓度的Mo作用后,形态发生异常,出现畸形,细胞活力下降,细胞抗氧化能力会受到抑制从而导致了氧化损伤的加剧,同时还抑制了Vimentin、α-actin和β-actin的mRNA和蛋白的表达。This experiment was conducted to investigate the effect of molybdenum(Mo)on oxidative damage and cytoskeleton in skeletal muscle epithelial cells of mouse.Used the primary skeletal muscle epithelial cells of mouse as materials,and cultured for 24 h in medium with Mo concentrations of 0(groupⅠ),0.1(groupⅡ),0.2(groupⅢ),0.4(groupⅣ),and 0.8 mmol/L(groupⅤ),respectively.Inverted microscope for cell morphology changes,methylthio tetrazole(MTT)method for cell viability,colorimetric assay for cell antioxidant capacity and oxidative damage,and quantitative real-time PCR(qRT-PCR)and Western-blot for Vimentin,α-actin andβ-actin mRNA and protein expression.The results showed as follows:1)the morphology of cells in groupsⅠandⅡwas normal,cell gaps in groupsⅢ,ⅣandⅤwere increased,cells were swollen,and cells in groupⅤshowed obvious deformity.2)Compared with the groupⅠ,the viability in skeletal muscle epithelial cells of mouse in groupsⅡ,Ⅲ,ⅣandⅤwas significantly decreased(P<0.01).3)Compared with the groupⅠ,the superoxide dismutase activity and total antioxidant capacity in skeletal muscle epithelial cells of mouse in groupsⅡ,Ⅲ,ⅣandⅤwere significantly decreased(P<0.01),and the malondialdehyde content was significantly increased(P<0.01).4)Compared with the groupⅠ,the mRNA relative expressions of Vimentin,α-actin andβ-actin in skeletal muscle epithelial cells of mouse in groupsⅡandⅢwere significantly increased(P<0.05),and that in groupⅤwas significantly decreased(P<0.05);the protein relative expressions of Vimentin,α-actin andβ-actin in skeletal muscle epithelial cells of mouse in groupsⅢ,ⅣandⅤwere significantly decreased(P<0.05).In conclusion,after the skeletal muscle epithelial cells of mouse treated with high concentration of Mo,the morphology is abnormal and deformity,cells viability is decrease,the anti-oxidation ability is inhibited and the oxidative damage is aggravated,at the same time it can also inhibit the mRNA and protein expression of Vimentin,α-actin andβ-a

关 键 词：钼 骨骼肌上皮细胞 氧化损伤 细胞骨架蛋白 

分 类 号：S816.72[农业科学—饲料科学]