有氧运动通过TLR4/miR-223/NLRP3信号通路轴介导CUMS抑郁小鼠海马炎症反应  

Aerobic Training Inhibits Hippocampal Inflammation by Activating the Hippocampus TLR4/miR223/NLRP3 Signaling Pathway Axis in Mice with CUMS Depression

作　　者：屈红林 谢军[2] 陈嘉勤[1] 刘瑞莲[2] 汤长发[1] 陈伊琳[1] 陈伟[1] 李娣[1] 彭琪 陈锐 QU Honglin;XIE Jun;CHEN Jiaqin;LIU Ruilian;TANG Changfa;CHEN Yilin;CHEN Wei;LI Di;PENG Qi;CHEN Rui(Hunan Normal University,Changsha 420012,China;Yichun University,Yichun 336000,China)

机构地区：[1]湖南师范大学体育学院湖南省体适能与运动康复重点实验室,湖南长沙420012 [2]宜春学院体育科学研究所,江西宜春336000

出　　处：《体育科学》2019年第2期39-50,共12页China Sport Science

基　　金：江西省卫计委科技项目(20171862);湖南省教育厅科技重点项目(1293)

摘　　要：目的:探讨有氧运动激活慢性不可预见性应激(CUMS)抑郁小鼠海马TLR4/miR-223/NLRP3信号通路改善海马功能的作用。方法:8周龄雄性C57BL/6小鼠60只,随机数字法分为空白对照组(CG)、抑郁模型组(MG)、模型运动组(ME)、TLR4抑制剂组(TG)和TLR4抑制剂+运动组(TE),每组12只。除空白对照组外的其他小鼠均进行为期28天的慢性应激性抑郁造模。造模后进行神经行为学评定,造模成功的ME、TE组小鼠适应性运动1周后进行为期8周的中等强度跑台运动,抑制剂组进行为期4周的TAK-242腹腔注射。训练8周后当日评定各组小鼠的神经行为学改变,次日取样,ELISA法测定血清IL-1β和IL-10含量,尼氏染色检测海马尼氏体形态,海马组织进行m RNA和miRNA的高通量测序及关联分析,免疫组化法测定TLR4、IL-1β、IL-10和NF-κB蛋白阳性表达区域面积,RT-PCR检测海马NLRP3、TLR4、IL-1β、IL-10、NF-κB和mi R-223的表达,WesternBlot检测海马TLR4、IL-10和IL-1β蛋白定性表达。结果:CUMS抑郁小鼠神经行为学功能显著下降,血液IL-1β含量明显增多,海马尼氏体核固缩严重。8周的有氧运动能够有效改善CUMS抑郁小鼠海马功能,降低血清IL-1β含量,显著减少海马组织尼氏体核固缩,降低小鼠的绝望行为,增加求生欲望。miRNA与m RNA高通量测序及关联分析结果显示,mi R-223与NLRP3存在靶向调控作用,且受运动干预的影响。ME组小鼠NLRP3、TLR4、IL-1β、NF-κB等炎症相关因子的表达下调,抗炎因子IL-10表达增强,miR-223表达增强。抑郁小鼠TLR4被抑制后,下游转录子NF-κB表达降低,mi R-223表达增强,NLRP3蛋白表达降低,炎性作用下降。TE组小鼠也观察到一致的抗炎症效果。结论:有氧运动可显著降低CUMS抑郁小鼠海马TLR4表达,激活TLR4/miR-223/NLRP3通路轴,改善抑郁小鼠海马功能,拮抗抑郁小鼠海马组织炎症,促进损伤的海马组织修复。Objective:The purpose of this study was to explore the effects of aerobic training on the hippocampal inflammation by activating the hippocampal TLR4/miR-223/NLRP3 signal pathway in mice with chronic unpredictable mild stress depression(CUMS).Methods:Sixty 8-week-old male C57BL/6 mice were divided into control group(CG),CUMS model group(MG),CUMS model plus exercise group(ME),TLR4 inhibitor group(TG)and TLR4 inhibitor plus exercise group(TE)(n=12).Except CG,the CUMS was molded in mice of the other four groups by suffering from chronic stress for 28days.Both ME group and TE group were trained for 8 weeks by running on a treadmill in moderate-intensity.TG group and TE group were received intraperitoneal injection of TAK-242,which was an inhibitor of TLR4,for 4 weeks.The neurobehavioral scores were tested after the 8-week training for all mice.Then,the samples were collected the day after their last training.The level of blood IL-1βand IL-10 were measuredby ELISA kit,the morphology of Nissl body in hippocampus were detected by Nissl staining,the positive expression areasof TLR4,IL-1β,IL-10 and NF-κB protein weredetected by immunohistochemistry,the gene expression levels of hippocampus NLRP3,TLR4,IL-1β,IL-10,NF-κB and miR-223 were examined by RT-PCR,and the protein expression levels of hippocampus TLR4,IL-1βand IL-10 were tested by Western Blot.Results:The neurobehavioral function of CUMS mice was significantly decreased,the inflammatory factors such as IL-1βin blood were significantly increased,and the Nissl body nuclear pyknosis were observed.8-week aerobic training improved the hippocampus function in CUMS micesignificantly,it also reduced the level of IL-1βin blood and Nissl body nuclear pyknosis the desperate behaviorswere reduced and thedesire to survive was increased in the mice.High-throughput sequencing and correlation analysis of miRNA and mRNA showed that there was targeting regulatory relations between miR-223 and NLRP3,which was influencedby exercise intervention.The expressions of inflammatory fa

关 键 词：抑郁症 慢性不可预见性应激 miR-223 有氧运动 海马 TLR4/miR-223/NLRP3 

分 类 号：G804.7[文化科学—运动人体科学]