亚硒酸钠和硒代蛋氨酸对马骨骼肌卫星细胞氧化损伤的缓解作用及线粒体功能的影响

doi:10.16431/j.cnki.1671-7236.2025.10.007

摘要/Abstract

摘要： 【目的】研究不同浓度亚硒酸钠和硒代蛋氨酸对H₂O₂诱导的马骨骼肌卫星细胞氧化损伤的缓解作用及线粒体功能的影响，为硒作为抗氧化剂缓解马运动氧化应激提供参考依据。【方法】本试验利用600 μmol/L H₂O₂诱导马骨骼肌卫星细胞氧化应激模型，在饥饿培养基培养6 h后，换为含有不同浓度亚硒酸钠(10、20、50、100、150、200 nmol/L)和硒代蛋氨酸(10、20、50、100、150、200 nmol/L)的溶液培养24 h，再利用600 μmol/L H₂O₂诱导马骨骼肌卫星细胞6 h，收集样品。使用CCK-8试剂盒检测细胞活性，用抗氧化酶试剂盒检测抗氧化酶活性指标，实时荧光定量PCR检测抗氧化基因表达量，用线粒体压力试剂盒检测骨骼肌卫星细胞线粒体能量代谢，评价不同浓度亚硒酸钠和硒代蛋氨酸对氧化损伤的缓解作用。【结果】100 nmol/L亚硒酸钠组和150 nmol/L硒代蛋氨酸组细胞相对活力显著高于损伤组(P＜0.05)，与对照组无显著差异(P＞0.05)，总抗氧化能(T-AOC)与抗氧化酶活性显著高于损伤组和对照组(P＜0.05)。细胞中丙二醛(MDA)含量随亚硒酸钠浓度的增加呈先降低后增高的变化趋势，100 nmol/L亚硒酸钠组含量最低，显著低于对照组和损伤组(P＜0.05)；150 nmol/L硒代蛋氨酸组细胞中MDA含量显著低于对照组和损伤组(P＜0.05)。100 nmol/L亚硒酸钠和150 nmol/L硒代蛋氨酸组细胞中原癌基因(RELA)、谷胱甘肽过氧化物酶1(GPX1)、超氧化物歧化酶1(SOD1)基因表达量极显著高于对照组与损伤组(P＜0.01)，硫氧还蛋白还原酶 1(TRXR1)基因表达量极显著高于损伤组(P＜0.01)，150 nmol/L硒代蛋氨酸细胞中TRXR1基因表达量极显著高于对照组(P＜0.01)。150 nmol/L硒代蛋氨酸组细胞中核因子E2相关因子2(Nrf2)基因表达量显著高于损伤组(P＜0.05)，RELA、GPX1、TRXR1和SOD1基因表达量极显著高于对照组(P＜0.01)，基础呼吸值、ATP氧消耗值、最大呼吸值分别为32.28、21.70和45.01 pmol/min，显著高于损伤组(P＜0.05)，分别增加了25.03%、30.22%和25.59%，而100 nmol/L亚硒酸钠组细胞耗氧率和线粒体功能参数与损伤组无显著差异(P＞0.05)。【结论】亚硒酸钠和硒代蛋氨酸通过激活Nrf2下游信号通路，上调抗氧化基因的表达，进而提高超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX)活性，减少MDA的产生，缓解氧化损伤，本试验条件下100 nmol/L亚硒酸钠和150 nmol/L硒代蛋氨酸缓解效果较好。相比无机的亚硒酸钠，有机的硒代蛋氨酸还可提高细胞的线粒体能量代谢。

关键词: 马; 骨骼肌卫星细胞; 亚硒酸钠; 硒代蛋氨酸; 氧化损伤

Abstract: 【Objective】 This study aimed to investigate the effects of different concentrations of sodium selenite and selenomethionine on H₂O₂-induced oxidative damage and mitochondrial function in horse skeletal muscle satellite cells,and provide a reference for selenium as an antioxidant to alleviate oxidative stress during exercise in horses.【Method】 In this study,the oxidative stress model of horse skeletal muscle satellite cells was induced by 600 μmol/L H₂O₂.After 6 h of starvation medium culture,different concentrations of sodium selenite (10,20,50,100,150 and 200 nmol/L) and selenomethionine (10,20,50,100,150 and 200 nmol/L) were used to culture for 24 h.Then,horse skeletal muscle satellite cells were induced with 600 μmol/L H₂O₂ for 6 h.Finally,samples were collected.CCK-8 kit was used to detect the cell viability,antioxidant enzyme kit was used to detect the indices of antioxidant enzyme activity,Real-time quantitative PCR was used to detect the expression of antioxidant genes,and mitochondrial stress kit was used to detect the mitochondrial energy metabolism of skeletal muscle satellite cells to evaluate the mitigation effect of different concentrations of sodium selenite and selenomethionine on oxidative damage.【Result】 The relative cell viability of 100 nmol/L sodium selenite group and 150 nmol/L selenomethionine group was significantly higher than that of injury group (P＜0.05),there was no significant difference compared with control group(P＞0.05),the total antioxidant capacity (T-AOC) and antioxidant enzyme activity were significantly higher than those of injury and control groups (P＜0.05).The content of malondialdehyde (MDA) in cells decreased first and then increased with the increase of sodium selenite concentration,it was the lowest in 100 nmol/L sodium selenite group,which was significantly lower than those in control and injury groups (P＜0.05).The content of MDA in 150 nmol/L selenomethionine group was significantly lower than that in control and injury groups (P＜0.05).The expressions of RELA,glutathione peroxidase 1 (GPX1),superoxide dismutase 1 (SOD1) genes in 100 nmol/L sodium selenite and 150 nmol/L selenomethione groups were extremely significantly higher than those in control and injury groups (P＜0.01),the expression of thioredoxin reductase 1 (TRXR1) gene was extremely significantly higher than that in injury group (P＜0.01),and the expression of TRXR1 gene in 150 nmol/L selenomethione group was extremely significantly higher than that in control group (P＜0.01).The expression of nuclear factor E2-related factor 2 (Nrf2) gene in 150 nmol/L selenomethionine group was significantly higher than that in injury group (P＜0.05).The expression of RELA,GPX1,TRXR1 and SOD1 genes was extremely significantly higher than that in control group (P＜0.01).The basal respiration value,ATP oxygen consumption value and maximum respiration value were 32.28,21.70 and 45.01 pmol/min,respectively,which were significantly higher than those in injury group (P＜0.05),increased by 25.03%,30.22% and 25.59%,respectively.However,there was no significant difference in oxygen consumption rate and mitochondrial function parameters between 100 nmol/L sodium selenite group and injury group (P＞0.05).【Conclusion】 Sodium selenite and selenomethionine could up-regulate the expression of antioxidant genes by activating the downstream signaling pathway of Nrf2,thereby increasing the activities of superoxide dismutase (SOD),catalase (CAT) and glutathione peroxidase (GPX),reducing MDA production and alleviating oxidative damage.Under this test condition,100 nmol/L sodium selenite and 150 nmol/L selenomethionine had the best remission effect.In addition,compared with the inorganic sodium selenite,organic selenomethionine could also improve mitochondrial energy metabolism.

Key words: horse; skeletal muscle satellite cells; sodium selenite; selenomethionine; oxidative damage

中图分类号:

S816.72

张心壮, 刘雨蒙, 高星, 王晓阳, 尹艺旋, 芒来. 亚硒酸钠和硒代蛋氨酸对马骨骼肌卫星细胞氧化损伤的缓解作用及线粒体功能的影响[J]. 中国畜牧兽医, 2025, 52(10): 4603-4613.

ZHANG Xinzhuang, LIU Yumeng, GAO Xing, WANG Xiaoyang, YIN Yixuan, MANG Lai. Effects of Sodium Selenite and Selenomethionine on the Mitigation of Oxidative Damage and Mitochondrial Function in Equine Skeletal Muscle Satellite Cells[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4603-4613.

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