孕期运动调节高血压大鼠子代血管平滑肌表型的Ppargc1α去甲基化机制  

Gestational exercise regulates vascular smooth muscle phenotype in offspring of hypertensive rats through Ppargc1αdemethylation

作　　者：陈渝[1,3] 李珊珊 张严焱[1] 刘晨喆 李丽 石丽君[1,2] CHEN Yu;LI Shan-shan;ZHANG Yan-yan;LIU Chen-zhe;LI Li;SHI Li-jun(Department of Exercise Physiology,Beijing 100084,China;Key Laboratory of Physical Fitness and Exercise,Ministry of Education,Beijing Sport University,Beijing 100084,China;College of Physical Education,Chongqing University,Chongqing 400044,China)

机构地区：[1]北京体育大学运动生理教研室,北京100084 [2]北京体育大学运动与体质健康教育部重点实验室,北京100084 [3]重庆大学体育学院,重庆400044

出　　处：《中国病理生理杂志》2021年第12期2122-2130,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.32071174,No.31771312);中央高校基本科研业务费专项资金资助项目(No.2020049)。

摘　　要：目的:探讨孕期运动是否抑制高血压大鼠子代血管重构及其可能的表观遗传机制。方法:选用自发性高血压大鼠(SHR)及其正常血压对照大鼠(WKY),同品系进行配种,见栓且阴道涂片见精子确定为配种成功,记为妊娠第1天。孕鼠随机分为安静(p-WKY-SED和p-SHR-SED)组和运动(p-WKY-EX和p-SHR-EX)组,每组25只。运动组进行无负重游泳运动(每天60 min,每周6 d,至妊娠第19天)。每天监测孕鼠进食量和体重;妊娠第21天,一部分孕鼠剖腹取胎鼠监测胎鼠体重和胎盘效率,另一部分孕鼠待自然分娩;雄性子代3月龄时,进行尾动脉无创血压监测,采用HE染色观察肠系膜动脉形态,采用RT-qPCR和Western blot检测肠系膜动脉过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC1α)的mRNA和蛋白水平及TET2蛋白水平,亚硫酸氢盐测序和ELISA分别检测肠系膜动脉Ppargc1α基因(编码PGC1α)甲基化水平和5-羟甲基胞嘧啶(5hmC)水平。结果:孕期运动抑制SHR子代胎盘效率下降(P<0.05),显著降低SHR子代的舒张压和平均动脉压(P<0.05),显著减小SHR子代的血管中膜厚度/内径比值(P<0.01),抑制血管平滑肌收缩表型向合成表型转换(P<0.05),显著上调SHR子代肠系膜动脉减少的PGC1αmRNA和蛋白表达,抑制Ppargc1α基因甲基化(P<0.05);与去甲基化密切相关的TET2蛋白(P<0.05)和5hmC(P<0.01)在SHR子代肠系膜动脉中显著降低,孕期运动显著增加二者水平(P<0.05)。结论:孕期运动可通过增加TET2介导的Ppargc1α基因启动子区去甲基化,抑制高血压大鼠子代成年后血管平滑肌由收缩表型向合成表型转换,缓解其血管重构。AIM:To investigate whether gestational exercise inhibits vascular remodeling in offspring of hypertensive rats and its underlying epigenetic mechanism.METHODS:Spontaneously hypertensive rats(SHR)and normotensive Wistar-Kyoto(WKY)rats were used in current study.After line breeding,observation of vaginal plug and spermatozoa in the vaginal smear was regarded as successful mating and recorded as gestational day 1.Pregnant rats were randomly divided into sedentary(p-WKY-SED and p-SHR-SED)group and exercise(p-WKY-EX and p-SHR-EX)group,with 25 rats in each group.The rats in exercise groups were performed a swimming training(60 min/day,6 day/week,till to gestational day 19).The food intake and body weight of pregnant rats were calculated every day.The fetuses were collected from some rats on the 21 st day of pregnancy to detect the body weight and placental weight.Blood pressure of 3-month-old male offspring was measured by caudal artery tail-cuff method.The morphological changes of the mesenteric artery were observed by HE staining.RT-qPCR and Western blot were used to detect the mRNA and protein levels of peroxisome proliferator-activated receptorγcoactivator 1α(PGC1α)and the TET2 protein level in the mesenteric artery.Bisulfite sequencing PCR and ELISA were used to detect the methylation rate of Ppargc1αgene(encoding PGC1α)and the level of 5-hydroxymethylcytosine(5 hmC)in the mesenteric artery,respectively.RESULTS:Gestational exercise significantly increased the placental efficiency in p-SHR-EX group compared with p-SHR-SED group(P<0.05).Diastolic blood pressure and mean arterial pressure of offspring in p-SHR-EX group were significantly reduced compared with p-SHR-SED group(P<0.05).The ratio of medial layer thickness to lumen diameter were decreased and the switch from contractile phenotype to synthetic one were inhibited by gestational exercise(P<0.01).Gestational exercise significantly increased PGC1αmRNA and protein,and inhibited the methylation rate of Ppargc1αgene in SHR offspring(P<0.05).The levels of T

关 键 词：孕期运动 自发性高血压大鼠 TET2蛋白 Ppargc1α基因 血管平滑肌表型 

分 类 号：R714.246[医药卫生—妇产科学] R363.2[医药卫生—临床医学]