有氧运动通过调节FoxO3a磷酸化及亚细胞定位减轻小肠缺血再灌注损伤  

Aerobic Exercise Attenuates Intestinal Ischemia/Reperfusion Injury by Regulating Phosphorylation and Subcellular Localization of FoxO3a

作　　者：韩雪[1] 张新敏[2] 陈岱莉[3] 曹君 黄晓雷[3] Han Xue;Zhang Xinmin;Chen Daili(Department of Anesthesiology,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510120,China;Department of Anesthesiology,First Hospital of Jilin University,Changchun 130061,China;Department of Anesthesiology,Shenzhen Maternity&Child Healthcare Hospital,Southern Medical University,Shenzhen 518028,China)

机构地区：[1]中山大学孙逸仙纪念医院麻醉科,广州510120 [2]吉林大学第一医院麻醉科,长春130061 [3]南方医科大学附属深圳市妇幼保健院麻醉科,深圳518028

出　　处：《华中科技大学学报（医学版）》2023年第4期439-444,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81900578);深圳市科技计划项目(No.JCYJ20180306173050592);广东省基础研究与应用基础研究项目(No.2019A1515011101);广州市科技计划项目(No.202201020261);广东省医学科学技术研究基金项目(No.B2022175)。

摘　　要：目的探讨有氧运动通过调控叉头框家族蛋白O3a(forkhead box protein O3a,FoxO3a)活性在C57BL/6小鼠小肠缺血/再灌注(II/R)损伤中的保护作用及机制。方法6~8周龄雄性C57BL/6小鼠随机分为静息+假手术(SS)组、静息+II/R(SI)组、游泳训练+II/R(TI)组和游泳训练+II/R+FoxO3a磷酸化激活剂SC79(TSC)组。TI和TSC组进行为期5周的游泳训练,所有小鼠于训练结束后3 d行II/R术,SS组不夹闭血管,TSC组小鼠于再灌注前1 h腹腔注射0.04 mg/g SC79。再灌注2 h后处死小鼠取标本,苏木精-伊红染色观察小肠组织病理改变,DHE染色检测活性氧(reactive oxygen species,ROS)水平。同时检测小肠组织丙二醛(malondialdehyde,MDA)含量以及过氧化氢酶(catalase,CAT)和超氧化物歧化酶(superoxide dismutase,SOD)活性。Western blot法检测Ser253磷酸化叉头框家族蛋白O3a(P-FoxO3a^(Ser253))、FoxO3a表达。qRT-PCR检测抗氧化基因锰超氧化物歧化酶(Manganese superoxide dismutase,MnSOD)和过氧化物酶体增殖物活化受体γ共激活因子1α(peroxisome proliferator-activated receptorγcoactivator 1α,PGC1α)的表达。结果II/R前行有氧运动干预,通过发挥抗氧化应激作用,减轻II/R损伤。与SI组小鼠相比,TI组小鼠小肠组织细胞质内P-FoxO3a^(Ser253)表达水平下调(P<0.01),细胞核内FoxO3a含量提高(P<0.01),抗氧化基因MnSOD和PGC1α的表达增加(P<0.05和P<0.01),小肠组织ROS生成减少(P<0.01),氧化应激产物MDA含量明显下降(P<0.05),抗氧化酶CAT和SOD含量增加(P<0.05和P<0.01);再灌注前腹腔注射SC79后,上述有氧运动减轻II/R损伤作用被抵消。结论有氧运动通过调节FoxO3a磷酸化及亚细胞定位减轻C57BL/6小鼠II/R损伤。Objective To explore the protective effect and mechanism of aerobic exercise on intestinal ischemia/reperfusion(II/R)injury in C57BL/6mice by regulating the activity of forkhead box protein O3a(FoxO3a).Methods Male C57BL/6mice,aged 6~8weeks,were randomly divided into Sedentary+Sham(SS)group,Sedentary+II/R(SI)group,Swim training+II/R(TI)group and Swim training+II/R+SC79,a FoxO3aphosphorylation activator,(TSC)group.Mice in TI and TSC groups recevied swim training for 5weeks.All of the mice underwent II/R surgery 3days after training while the superior mesenteric artery was not clamped in SS group.Mice in TSC group were intraperitoneally injected with 0.04mg/g SC79one hour before reperfusion.Mice were sacrificed for sample collection two hours after reperfusion.Pathological changes of the intestine mucosa were observed by HE staining,level of reactive oxygen species(ROS)by DHE staining,while the content of malondialdehyde(MDA)and the activity of catalase(CAT)and superoxide dismutase(SOD)were detected in intestinal tissues.Levels of PFoxO3a^(Ser253) and FoxO3awere tested by Western blotting and expression of antioxidative genes,manganese superoxide dismutase(MnSOD)and peroxisome proliferator-activated receptorγcoactivator 1α(PGC1α)by qRT-PCR.Results Aerobic exercise intervention before II/R surgery has the potential of antioxidative stress to alleviating II/R injury.Compared to SI group,the cytoplasmic level of P-FoxO3a^(Ser253) was downregulated(P<0.01),the nuclear level of FoxO3awas upregulated(P<0.01),the expression of antioxidant genes MnSOD and PGC1αwere increased(P<0.05,P<0.01),the generation of intestinal ROS reduced(P<0.01),the content of oxidative stress product MDA was significantly decreased(P<0.05)while antioxidant enzymes CAT and SOD were increased(P<0.05,P<0.01).However,the protective effects were abolished by intraperitoneal injection of SC79before reperfusion.Conclusion Aerobic exercise alleviates II/R injury in C57BL/6mice by regulating FoxO3a phosphorylation and subcellular localization.

关 键 词：有氧运动 叉头框家族蛋白O3a 磷酸化 氧化应激 肠缺血再灌注损伤 

分 类 号：R656.7[医药卫生—外科学]