高强度间歇训练对增龄大鼠脂...分泌表型的影响及其机制探讨_马松.pdf

高强度间歇训练对增龄大鼠脂肪组织衰老相关分泌表型的影响及其机制探讨Effects and Mechanisms of High-Intensity Intervals Training on Senescence-Associated Secretory Phenotype in Adipose Tissue of Aging Rats and Mechanisms马松1，李方晖1，2*，吴大帅1，谢天1，王静1，乔一波1MA Song1，LI Fanghui1，2*，WU Dashuai1，XIE Tian1，WANG Jing1，QIAO Yibo1摘要：目的：探讨高强度间歇训练（HIIT）对增龄大鼠脂肪组织衰老相关分泌表型（SASP）的影响及机制。方法：将18月龄雌性大鼠分为安静对照组（SED）、持续性耐力组（MICT）和HIIT组，每组12只。HIIT组起始速度为15 m/min，时间为4 min；之后进行高强度训练，速度为25 m/min，运动1 min，依次交替9个循环。MICT组以17 m/min速度运动45 min，持续8个月。免疫组化观测内脏脂肪-半乳糖苷酶，蛋白免疫印迹和RT-qPCR检测脂肪组织p16-INK4a、p53、脂联素（Adiponectin）、瘦素（Leptin）、白介素-6（IL-6）、白介素-1（IL-1）、肿瘤坏死因子-（TNF-）和单核细胞趋化蛋白-1（MCP-1）表达，蛋白免疫印迹检测核转录因子-B（NF-B）、p38丝裂原活化蛋白激酶（p38MAPK）、蛋白激酶B（AKT）、核转录因子-B抑制蛋白 （IkB）和 c-Jun N 端激酶（JNK）总蛋白及其磷酸化水平，蛋白免疫荧光观察p16-INK4a、p-NF-B和p-lkB蛋白含量。结果：与SED和MICT组相比，HIIT组-半乳糖苷酶染色阳性脂肪细胞数（P0.05）及IL-6、IL-1、瘦素、TNF-、p16-INK4a蛋白和mRNA表达显著降低（P0.01），脂联素的蛋白（P0.05）和 mRNA 表达（P0.01）显著升高；HIIT 和MICT组脂肪组织p53蛋白和mRNA表达均低于SED组（P0.01），HIIT组p-IkB、p-JNK、p-NF-B、p-p38MAPK、p-IkB/p-IkB、p-JNK/JNK、p-NF-B/NF-B、p-p38MAPK/p38MAPK均低于SED和MICT组（P0.01），HIIT组p-AKT和p-AKT/AKT高于SED和MICT组（P0.01）。相关性分析显示，p-NF-B/NF-B 与 p16-INK4a（r=0.94，P0.01）、IL-6（r=0.77，P0.05）、IL-1（r=0.74，P0.05）、TNF-（r=0.82，P0.01）、瘦素（r=0.92，P0.01）呈正相关，p-AKT/AKT与p-IkB/IkB（r=-0.89，P0.01）、p-JNK/JNK（r=-0.95，P0.01）、p-p38MAPK/p38MAPK（r=-0.90，P0.01）和p-NF-B/NF-B（r=-0.91，P0.01）呈负相关。结论：HIIT比MICT能更有效地抑制内脏脂肪组织SASP分泌，该过程可能与AKT通路的激活及NF-B、p38MAPK和JNK通路的抑制有关。关键词：高强度间歇训练；增龄；脂肪组织；衰老相关分泌表型 Abstract:Objective:To investigate the effects and mechanism of high-intensive interval training(HIIT)on senescence-associated secretory phenotype(SASP)in adipose tissue of aging rats.Methods:36 female Sprague Dawley rats at 18 months old were selected randomly and divided into three groups:Sedentary control(SED),moderate-intensity continuous training(MICT),and HIIT groups,12 rats per group.HIIT started with a speed of 15 m/min and lasted for 4 min.After that,the high-intensity training speed was 25 m/min,and the time was 1 min.Then,9 cycles were successively alternated.The MICT had a speed of 17 m/min and a time of 45 min.The exercise lasted for eight months.Immunohistochemistry was used to examine the contents of beta-galactosidase in visceral adipocytes.Expressions of p16-INK4a,p53,adiponectin,leptin,interleukin-6(IL-6),interleukin-1(IL-1),and monocyte chemoattractant protein-1(MCP-1)in visceral adipose tissue was detected by Western Blot and RT-PCR.In addition,Western Blot was performed to detect the phosphorylation levels of NF-B,p38MAPK,AKT,IkB,and JNK.More中国体育科技2023 年（第59卷）第2期CHINA SPORT SCIENCE AND TECHNOLOGYVol.59,No.2,25-34,2023文章编号：1002-9826（2023）02-0025-10DOI：10.16470/j.csst.2021126基金项目：国家自然科学基金项目（31971099）；广 东 省 省 级 科 技 计 划 项 目（2015A020219015）第一作者简介：马松（1996-），男，在读硕士研究生，主要研究方向为运动与抗衰老，E-mail：。*通信作者简介：李方晖（1983-），男，教授，博士，博士研究生导师，主要研究方向为运动抗衰老机制，E-mail：。作者单位：1.南京师范大学，江苏 南京 210023；2.肇庆学院，广东 肇庆 5260611.Nanjing Normal University,Nanjing 210023,China;2.Zhaoqing University，Zhaoqing 526061，China.25中国体育科技2023年（第59卷）第2期over,the protein contents of p16-INK4a,p-NF-B,and p-IkB were observed by immunofluorescence.Results:Compared with the SED and MICT groups,the number of-galactosidase staining positive adipocytes,and the protein and mRNA expression of IL-6,IL-1,leptin,TNF-,and p16-INK4a in the HIIT group were significantly decreased(P0.01),and the expression of adiponectin protein(P0.05)and mRNA expression were significantly increased(P0.01).The expression of p53 protein and mRNA in HIIT and MICT group was lower than that in SED group(P0.01).Western blot showed that the p-IkB,p-JNK,p-NF-B,p-p38MAPK,p-IkB to p-IkB ratio,p-JNK to JNK ratio,p-NF-B to NF-B ratio,and p-38MAPK to p38MAPK ratio in the HIIT group were lower than those in the SED and MICT groups(P0.01),and the p-AKT and p-AKT to AKT ratio were higher than those in the SED and MICT groups(P0.01).Pearson correlation analysis showed that p-NF-B to NF-B ratio was positively correlated with p16-INK4a(r=0.94,P0.01),IL-6(r=0.77,P0.05),IL-1(r=0.74,P0.05),TNF-(r=0.82,P0.01),and leptin(r=0.92,P0.01),and p-AKT to AKT ratio was negatively correlated with p-IkB to IkB ratio(r=-0.89,P0.01),p-JNK to JNK ratio(r=-0.95,P0.01),p-p38MAPK to p38MAPK ratio(r=-0.90,P0.01),and p-NF-B to NF-B ratio(r=-0.91,P0.01).Conclusions:HIIT exhibited greater beneficial effects on inhibiting SASP secretion in visceral adipose tissue than MICT,and this process may be related to activation of AKT signaling pathway,and inhibition of NF-B,JNK,and p38MAPK pathway.Keywords:high-intensity intervals training;aging;adipose;senescence-associated secretory phenotype中图分类号中图分类号：G804.2 文献标识码文献标识码：A脂肪组织是机体重要的内分泌器官（Kershaw et al.，2004）。在增龄性衰老过程中，脂肪组织分布、结构和功能将发生变化，衰老脂肪细胞可大量分泌由炎性因子、脂肪因子等一系列细胞因子组成的衰老相关分泌表型（senescence-associated secretory phenotype，SASP）（郭 慧 宁 等，2016；殷双丽 等，2018；Copp et al.，2010）。已有研究证实，衰老脂肪细胞释放 SASP 可加快机体衰老进程（Tam et al.，2020），其与衰老相关疾病的发生及发展密不可分（Zhu et al.，2019）。因此，调控脂肪组织 SASP 释放是延缓机体衰老的重要途经之一（Yang et al.，2019）。SASP的分泌过程依赖于核因子-B（nuclear factor-B，NF-B）通路的活化，通过调控炎性因子基因转录过程，可以促进NF-B表达（杨连伟，2019）。在衰老脂肪细胞中，IB和NF-B的