EGCG对热应激诱导猪骨骼肌卫星细胞凋亡的保护作用  

Protective effects of EGCG on heat stress-induced apoptosis in porcine skeletal muscle satellite cells

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作  者:杨豹 王倩 李银 蒋钦杨[1,2] 黄艳娜 YANG Bao;WANG Qian;LI Yin;JIANG Qin-yang;HUANG Yan-na(College of Animal Science and Technology,Guangxi University,Nanning,Guangxi 530004,China;Guangxi Key Laboratory of Animal Breeding,Disease Control and Prevention,Nanning,Guangxi 530004,China)

机构地区:[1]广西大学动物科学技术学院,广西南宁530004 [2]广西畜禽繁育与疾病防控重点实验室,广西南宁530004

出  处:《南方农业学报》2024年第10期3190-3198,共9页Journal of Southern Agriculture

基  金:国家自然科学基金项目(32360839,31760672);广西自然科学基金项目(2022GXNSFAA035525)。

摘  要:【目的】探究表没食子儿茶素没食子酸酯(EGCG)对热应激诱导猪骨骼肌卫星细胞凋亡的影响及其作用机制,为缓解猪热应激反应和促进猪肌肉生长提供理论参考。【方法】41.5℃处理2 d构建猪骨骼肌卫星细胞热应激模型,使用CCK-8法检测热应激和EGCG对细胞生长水平的影响,利用Hoechst 33258染色和流式细胞仪检测热应激和EGCG对细胞凋亡水平的影响,采用实时荧光定量PCR检测Bcl-2、BAX、Caspase-3、Caspase-9和Cytc基因相对表达量,探究EGCG对热应激细胞凋亡相关基因表达的影响,并通过Western blotting检测Bcl-2和BAX蛋白相对表达量及Bax/Bcl-2比值,探究EGCG对热应激细胞凋亡相关蛋白表达的影响。【结果】41.5℃处理2 d极显著降低猪骨骼肌卫星细胞生长水平(P<0.01,下同)并极显著提高细胞凋亡率;热应激能显著提高Caspase-3和Cytc基因及BAX蛋白的相对表达量(P<0.05,下同),极显著提高BAX和Caspase-9基因的相对表达量,极显著降低Bcl-2基因的相对表达量,显著降低Bcl-2蛋白的相对表达量。EGCG能缓解热应激诱导的猪骨骼肌卫星细胞生长水平下降和凋亡水平升高,并降低促凋亡相关基因BAX、Caspase-3、Caspase-9和Cytc及BAX蛋白的相对表达量,提高抗凋亡基因Bcl-2和蛋白Bcl-2的相对表达量。【结论】EGCG能缓解热应激诱导的猪骨骼肌卫星细胞生长水平下降,并通过调控Bcl-2、BAX、Caspase-3、Caspase-9和Cytc等抗/促凋亡相关基因及其蛋白的相对表达量来缓解热应激引起的细胞凋亡。【Objective】This study aimed to investigate the effects and mechanisms of epigallocatechin gallate(EGCG)on apoptosis of porcine skeletal muscle satellite cells induced by heat stress,providing theoretical reference for alleviating heat stress responses and promoting muscle growth in pigs.【Method】A heat stress model of porcine skeletal muscle sate-llite cells was established by treating cells at 41.5°C for 2 d.The effects of heat stress and EGCG on cell growth level were assessed using the CCK-8 assay.Hoechst 33258 staining and flow cytometry were used to detect effects of heat stress and EGCG on cell apoptosis levels.Followed by real-time fluorescence quantitative PCR to measure the relative expression levels of Bcl-2,BAX,Caspase-3,Caspase-9 and Cytc genes,exploring the effects of EGCG on apoptosis related gene expression under heat stress.Western blotting was performed to detect the relative expression levels of Bcl-2 and BAX proteins and the BAX/Bcl-2 ratio,investigating the effects of EGCG on apoptosis related protein expression under heat stress.【Result】Treatment at 41.5°C for 2 d extremely significantly decreased cell growth level(P<0.01,the same below)and extremely significantly increased apoptosis rate.Heat stress significantly increased the relative expression of Caspase-3 and Cytc genes and BAX protein(P<0.05,the same below)and extremely significantly elevated the relative expression of BAX and Caspase-9 genes,while extremely significantly reduced the relative expression of Bcl-2 gene and significantly reduced the relative expression of Bcl-2 protein.EGCG alleviated the decrease in cell growth level and the increase in apoptosis level of porcine skeletal muscle satellite cells induced by heat stress,reduced the relative expression of pro-apoptotic genes(BAX,Caspase-3,Caspase-9 and Cytc)and BAX protein,and increased the relative expression of the anti-apoptotic gene Bcl-2 and Bcl-2 protein.【Conclusion】EGCG mitigates the decline in growth level of porcine skele-tal muscle satellite cells i

关 键 词: EGCG 热应激 骨骼肌 卫星细胞 细胞凋亡 

分 类 号:S828.1[农业科学—畜牧学]

 

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