动物营养学报 >

2019 , Vol. 31 >Issue 12: 5664 - 5671

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.12.032

实验动物 Laboratory animal

钼对小鼠骨骼肌上皮细胞氧化损伤及细胞骨架的影响

  • 马淑浩 ,
  • 杨自军 ,
  • 杜伯强 ,
  • 李光照 ,
  • 张才 ,
  • 汪纪仓 ,
  • 王宏伟 ,
  • 孔涛
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  • 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, 洛阳 471000
马淑浩(1995-),男,河南柘城人,硕士研究生,研究方向为动物营养代谢与中毒病。E-mail:qazmsh@126.com

收稿日期: 2019-06-17

  网络出版日期: 2019-12-13

基金资助

国家自然科学基金项目（31040081）

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

  • MA Shuhao ,
  • YANG Zijun ,
  • DU Boqiang ,
  • LI Guangzhao ,
  • ZHANG Cai ,
  • WANG Jicang ,
  • WANG Hongwei ,
  • KONG Tao
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  • 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

Received date: 2019-06-17

  Online published: 2019-12-13

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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。

关键词： 钼; 骨骼肌上皮细胞; 氧化损伤; 细胞骨架蛋白

本文引用格式

马淑浩 , 杨自军 , 杜伯强 , 李光照 , 张才 , 汪纪仓 , 王宏伟 , 孔涛 . 钼对小鼠骨骼肌上皮细胞氧化损伤及细胞骨架的影响[J]. 动物营养学报, 2019 , 31(12) : 5664 -5671 . DOI: 10.3969/j.issn.1006-267x.2019.12.032

Abstract

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 (group Ⅰ), 0.1 (group Ⅱ), 0.2 (group Ⅲ), 0.4 (group Ⅳ), and 0.8 mmol/L (group Ⅴ), 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 and β-actin mRNA and protein expression. The results showed as follows:1) the morphology of cells in groups Ⅰ and Ⅱ was normal, cell gaps in groups Ⅲ, Ⅳ and Ⅴ were increased, cells were swollen, and cells in group Ⅴ showed obvious deformity. 2) Compared with the group Ⅰ, the viability in skeletal muscle epithelial cells of mouse in groups Ⅱ, Ⅲ, Ⅳ and Ⅴ was significantly decreased (P<0.01). 3) Compared with the group Ⅰ, the superoxide dismutase activity and total antioxidant capacity in skeletal muscle epithelial cells of mouse in groups Ⅱ, Ⅲ, Ⅳ and Ⅴ were significantly decreased (P<0.01), and the malondialdehyde content was significantly increased (P<0.01). 4) Compared with the group Ⅰ, the mRNA relative expressions of Vimentin, α-actin and β-actin in skeletal muscle epithelial cells of mouse in groups Ⅱ and Ⅲ were significantly increased (P<0.05), and that in group Ⅴ was significantly decreased (P<0.05); the protein relative expressions of Vimentin, α-actin and β-actin in skeletal muscle epithelial cells of mouse in groups Ⅲ, Ⅳ and Ⅴ were 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 and β-actin.

Key words： molybdenum; skeletal muscle epithelial cells; oxidative damage; cytoskeletal protein

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