n-3多不饱和脂肪酸通过抑制骨骼肌NLRP3炎症小体活性改善力竭运动小鼠疲劳  

n-3 polyunsaturated fatty acids improve fatigue by inhibiting NLRP3 inflammasome activity of skeletal muscle in exhaustive mice

作　　者：邓项元 张瑞良 陈帅 朱晓辉 糜漫天[1] 张婷[1] DENG Xiangyuan;ZHANG Ruiliang;CHEN Shuai;ZHU Xiaohui;MI Mantian;ZHANG Ting(Research Center for Nutrition and Food Safety,Chongqing Key Laboratory of Nutrition and Health,Chongqing Medical Nutrition Research Center,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,400038,China)

机构地区：[1]陆军军医大学(第三军医大学)军事预防医学系军队营养与食品卫生学教研室,重庆市医学营养研究中心,重庆市营养与健康重点实验室,重庆400038

出　　处：《陆军军医大学学报》2023年第24期2521-2529,共9页Journal of Army Medical University

摘　　要：目的揭示n-3多不饱和脂肪酸(n-3 polyunsaturated fatty acids,n-3 PUFAs)对力竭运动小鼠骨骼肌炎症和疲劳的作用及机制。方法选取40只鱼油灌胃的7周龄雄性C57BL/6J小鼠(外源性补充n-3 PUFAs)以及20只7周龄雄性Fat-1小鼠(内源性合成n-3 PUFAs),建立7 d力竭跑台运动小鼠模型。将C57BL/6J小鼠分为4组(每组10只):空白静息组(CON组)、空白运动组(EX组)、鱼油静息组(FO组)、鱼油运动组(FO+EX组);将Fat-1小鼠分为2组(每组10只):Fat-1静息组(FA组)、Fat-1运动组(FA+EX组)。利用全自动生化分析仪检测血清中尿素(urea,UREA)、葡萄糖(glucose,GLU)、肌酸激酶(creatine kinase,CK)、乳酸脱氢酶(lactate dehydrogenase,LDH);酶联免疫吸附试验测定血清IL-1β和IL-18含量;RT-qPCR检测腓肠肌IL-1β、IL-18、NLRP3、ASC、Caspase-1基因mRNA表达水平;蛋白免疫印迹法检测腓肠肌NLRP3和ASC蛋白表达水平;氧化应激试剂盒检测腓肠肌丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)以及肌糖原和肝糖原水平;HE染色观察腓肠肌组织形态结构变化。结果n-3 PUFAs降低了力竭运动小鼠血清UREA、CK、LDH水平(P<0.05),升高了GLU水平(P<0.05)。n-3 PUFAs降低了力竭运动小鼠血清IL-1β和IL-18水平(P<0.05)。n-3 PUFAs抑制了力竭运动小鼠骨骼肌NLRP3、ASC、Caspase-1、IL-1β和IL-18的mRNA表达水平(P<0.05),抑制了力竭运动小鼠骨骼肌NLRP3和ASC的蛋白表达水平(P<0.05)。n-3 PUFAs提高了力竭运动小鼠肝糖原和肌糖原含量(P<0.05)。n-3 PUFAs升高了力竭运动小鼠腓肠肌SOD、GSH-Px活性(P<0.05),降低了MDA含量(P<0.05)。结论n-3 PUFAs可有效提高小鼠抗氧化能力,改善力竭运动小鼠疲劳,其机制可能与n-3 PUFAs抑制力竭运动小鼠骨骼肌NLRP3炎症小体的过度激活有关。ObjectiveTo explore the effect and mechanism of n-3 polyunsaturated fatty acids(n-3 PUFAs)on skeletal muscle inflammation and fatigue in exhaustive exercise mice.MethodsUsing fish oil gavage to C57BL/6J mice for exogenous intervention of n-3 PUFAs and Fat-1 mice(endogenous synthesis of n-3 PUFAs),a 7-day exhaustive mice model was established.Forty C57BL/6J mice were divided into 4 groups(10 in each group):the blank resting group(CON group),the blank exercise group(EX group),the fish oil resting group(FO group)and fish oil exercise group(FO+EX group).Twenty Fat-1 mice were divided into 2 groups(10 in each group):the Fat-1 resting group(FA group)and the Fat-1 exercise group(FA+EX group).Serum levels of urinel UREA,glucose(GLU),creatine kinase(CK)and lactate dehydrogenase(LDH)were detected by automatic biochemical analyzer.Serum IL-1βand IL-18 levels were determined by enzyme-linked immunosorbent assay.The mRNA expression levels of IL-1β,IL-18,NLRP3,ASC and Caspase-1 in gastrocnemius muscle tissues were detected by RT-qPCR.The protein expression levels of NLRP3 and ASC in gastrocnemius muscle tissues were detected by Western blotting.Kits were used to detect the levels of malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),muscle glycogen and liver glycogen in gastrocnemius muscle.HE staining was used to observe the morphological and structural changes of gastrocnemius muscle.Resultsn-3 PUFAs decreased the levels of UREA,CK and LDH in exhaustive exercise mice(P<0.05),and increased the level of GLU(P<0.05).n-3 PUFAs decreased serum IL-1βand IL-18 levels in exhaustive exercise mice(P<0.05).n-3 PUFAs inhibited the mRNA expression levels of NLRP3,ASC,Caspase-1,IL-1βand IL-18 in skeletal muscle of exhaustive exercise mice(P<0.05),and the protein expression levels of NLRP3 and ASC in skeletal muscle of exhaustive exercise mice(P<0.05).n-3 PUFAs increased the content of liver and muscle glycogen in exhaustive exercise mice(P<0.05).n-3 PUFAs increased the activities of SOD and GSH-Px in gastroc

关 键 词：N-3多不饱和脂肪酸 力竭运动 NLRP3炎症小体 氧化应激 骨骼肌炎症 疲劳 

分 类 号：Q547[生物学—生物化学] R151.41[医药卫生—营养与食品卫生学] R872.7[医药卫生—公共卫生与预防医学]