摘要
目的观察雌马酚(equol,Eq)干预对棕榈酸(palmitic acid,PA)诱导的L6大鼠成肌细胞肌管萎缩的影响,并探讨其机制。方法体外培养L6细胞,以不同浓度的Eq、PA处理24 h,通过CCK-8法检测相对细胞活力确定Eq、PA干预的适宜浓度。将细胞分为对照组、模型组(PA:0.25 mmol/L)、干预组(PA:0.25 mmol/L+Eq:1μmol/L)和自噬抑制剂组(PA:0.25 mmol/L+Eq:1μmol/L+3-MA:1 mmol/L)。干预24 h后检测各组细胞葡萄糖摄取能力;HE染色观察细胞肌管生长情况;透射电镜观察细胞自噬小体数量;激光共聚焦显微镜观察经线粒体荧光探针标记的线粒体形态;Western blot检测MyHC、p62蛋白、LC3蛋白、MFN2和DRP1表达水平。结果干预24 h后,与对照组比较,模型组L6细胞葡萄糖消耗明显减低(P<0.05),肌管直径明显减小(P<0.05),自噬小体数量减少,线粒体裂变增加,MyHC、MFN2蛋白表达水平降低(P<0.05),而p62、DRP1表达水平及LC3-Ⅰ/LC3-Ⅱ比值增加(P<0.05);与模型组相比,干预组L6细胞葡萄糖消耗明显增加(P<0.05),肌管直径明显增大(P<0.05),自噬小体数量增加,线粒体裂变减少,MyHC、MFN2蛋白表达水平升高(P<0.05),而p62、DRP1表达水平及LC3-Ⅰ/LC3-Ⅱ比值降低(P<0.05);Eq对PA诱导的L6细胞葡萄糖消耗,肌管直径,自噬小体数量,线粒体裂变,MyHC、p62、MFN2、DRP1蛋白表达水平及LC3-Ⅰ/LC3-Ⅱ比值的影响可被3-MA有效抑制。结论Eq可有效改善PA诱导的L6细胞肌管萎缩,其部分机制与Eq通过促进自噬,减轻线粒体裂变有关。
Objective To obseve the effect of equol(Eg)intervention on palmitic acid(PA)-induced myoblast atrophy in rat myoblast cell line L6 and to explore its underlying mechanism.MethodsL6cells were dlvided into control group,model group(0.25 mmol/L PA),intervention group(0.25 mmol/LPA+1μmol/L Eq)and autophagy inhibitor(3-methyladenine,3-MA)group(0.25 mmol/L PA+1μmol/LEq+1 mmol/L 3-MA).Affer 24 h of intervention,glucose uptake capacity of ells was detected,muscle tubegrowth was observed by HE staining,autophagosome number was observed with transmision electionmicroscopy,and mitochondrial morphology labeled by fluorescent probe was oberved by laser confocalmicroscopy.The expresion levels of MyHC,p62,LC3,MFN2 and DRPI1 were detected by Westem hltting.ResultsCompared with the control goup,24 h of PA treatment resulted in significantly decreased glucose consumption(P<0.05),decreased muscle tube diameter(P<0.05),redhuced number of autophagosomes,increased mitochondrial fision,and reduced expression levels of MyHC and MFN2(P<0.05),and elevatedDRP1,p62 level and LC3-I/LC3-II ratio(P<0.05).While Eq interventon reserved above changesindhced by PA treatment(P<0.05).However,the effects of Eq on PA-induced glucose consumption,myotube diameter,autophagosome number,mitochondrial fision,protein levels of MyHC,p62,MFN2,DRP1 and LC3-I/LC3-II ratio in L6 cells could be effectively inhibited by 3-MA treatment.ConclusionEg efecivly ameliorates PA-induced muscle tube atophy in L6 cells,which may prtilly be related to E alleviating mitochondrial fision by promoting autophagy.
作者
于生财
倪向敏
卢亨
张贵明
王建
YU Shengcai;NI Xiangmin;LU Heng;ZHANG Guiming;WANG Jian(Department of Nutrition,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2023年第15期1604-1611,共8页
Journal of Army Medical University
基金
国家自然科学基金面上项目(81973040)。
