虾青素通过mTOR-ULK1自噬通路对大鼠运动性骨骼肌损伤影响的研究  

Effect of Astaxanthin on Exercise-induced Skeletal Muscle Damage in Rats through AMPK/mTOR/ULK1 Autophagy Pathway

作　　者：吴丽君[1,2] 王佳怡 毕翔宇 WU Lijun;WANG Jiayi;BI Xiangyu(School of P.E.,Shanxi University,Taiyuan 030006,Shanxi,China;Institute for Biomedicine and Health,Shanxi University,Taiyuan 030006,Shanxi,China)

机构地区：[1]山西大学体育学院,山西太原030006 [2]山西大学生物医药与大健康研究院,山西太原030006

出　　处：《山东体育学院学报》2024年第3期114-126,共13页Journal of Shandong Sport University

基　　金：山西省重点研发计划项目(编号:201803D31030)。

摘　　要：目的:基于AMPK/mTOR/ULK1通路探究虾青素对大鼠运动性骨骼肌损伤(EIMD)的影响。方法:健康雄性SD大鼠随机分为对照组(C)、运动组(E)、虾青素组(M)、运动+虾青素组(EM)、运动+AICAR(5-氨基-1-核糖基咪唑-4-甲酰胺磷酸盐组)组(EA)、运动+虾青素+AICAR组(EMA)、运动+虾青素+MHY1485组(EMM),每组10只。M、EM、EMA、EMM组灌胃虾青素;C、E组灌胃大豆油;EA、EMA组注射AICAR;EMM组注射MHY1485;E、EM、EA、EMA、EMM组进行力竭运动,其余组安静饲养。运动结束后,采用HE染色观察大鼠比目鱼肌的组织病理变化;ELISA法检测血清MDA、SOD、IL-6、IL-1β、CK和FINS含量;RT-PCR法检测比目鱼肌相关指标的mRNA表达;Western blot法检测比目鱼肌AMPK/mTOR/ULK1自噬通路磷酸化水平及凋亡相关蛋白表达;检测大鼠FBG和LA水平,计算ISI和HOMA-IR;此外,通过中介效应分析探究LA、MDA、IL-6和IL-1β在EIMD及ISI中的作用。结果:E组大鼠血清MDA、IL-6、IL-1β、CK、LA、FBG和FINS含量、HOMA-IR明显高于C组,SOD含量和ISI明显低于C组;大鼠比目鱼肌p-AMPK Thr172、p-mTOR Ser2448、p-ULK1 Ser555和p-ULK1 Ser757蛋白表达、P62基因和蛋白表达均明显高于C组,LC3-II蛋白表达、ILC3-II/LC3-I和BCL2/BAX明显低于C组。虽然,给力竭运动大鼠灌胃虾青素或注射AICAR可逆转上述改变,且与EA和EM组相比,EMA组逆转效果更加明显,但是,给力竭运动大鼠施加虾青素及MHY1485的双重干预则明显阻断了虾青素对EIMD的改善作用。此外,MDA、IL-6和IL-1β在LA导致的EIMD及ISI下降中发挥了部分中介效应。结论:力竭运动可造成EIMD,其发病机制涉及乳酸堆积、氧化应激、炎症反应及细胞凋亡等,此时,胰岛素抵抗增加,肌细胞对葡萄糖的摄取与利用减少,这将上调骨骼肌mTOR活性,抑制自噬通量,加剧骨骼肌损伤。而虾青素干预可调控mTOR-ULK1自噬通路以缓解EIMD,且虾青素与AICAR的联合作用对改善EIMD更有益。Objective:To explore the effects of astaxanthin on EIMD in rats based on the AMPK/mTOR/ULK1 pathway.Methods:Healthy male SD rats were randomly divided into a control(C)group,exercise(E)group,astaxanthin(M)group,exercise+astaxanthin(EM)group,exercise+AICAR(EA)group,exercise+astaxanthin+AICAR(EMA)group,and exercise+astaxanthin+MHY1485(EMM)group,with 10 rats in each group.Groups M,EM,EMA,and EMM were given astaxanthin;Groups C and E were given soybean oil;Groups EA and EMA were injected with AICAR;Group EMM was injected with MHY1485;Groups E,EM,EA,EMA,and EMM proceeded with exhaustive exercise,and the other groups were fed quietly.HE staining was used to observe the pathological changes in soleus tissues.The contents of MDA,SOD,IL-6,IL-1β,CK,and FINS in serum were detected by ELISA.mRNA expression of related indexes in soleus was detected by RT-PCR.Phosphorylation levels of the AMPK/mTOR/ULK1 autophagy pathway and apoptin expression in soleus were detected by Western blot.FBG and LA levels were detected,and ISI and HOMA-IR were calculated.The role of LA,MDA,IL-6,and IL-1βin EIMD and ISI was investigated by mediating effect analysis.Results:Compared with group C,the contents of MDA,IL-6,IL-1β,CK,LA,FBG,FINS,and HOMA-IR in the serum of group E were significantly increased,while SOD and ISI were significantly decreased;The expression levels of p-AMPK Thr172,p-mTOR Ser2448,p-ULK1 Ser555,p-ULK1 Ser757,and P62 were significantly increased,while the expression levels of LC3-II,ILC3-II/LC3-I and BCL2/BAX were significantly decreased.After gavage of astaxanthin or injection of AICAR,the above changes could be reversed.The reverse effect was more obvious in the EMA group than in the groups EA and EM.However,the dual intervention of astaxanthin and MHY1485 to exhaustive exercise rats blocked the protective effect of astaxanthin on EIMD.In addition,MDA,IL-6,and IL-1βplayed a partial mediating effect on EIMD and ISI caused by LA.Conclusion:Exhaustive exercise can cause EIMD,and the pathogenesis involves lactic acid accumulat

关 键 词：运动性骨骼肌损伤 虾青素 自噬 中介效应 

分 类 号：G804.2[文化科学—运动人体科学]