靶向调节哺乳动物雷帕霉素靶蛋白信号通路对心力衰竭小鼠调节性T细胞/辅助性T细胞17免疫自稳失衡的影响  

Effect of Targeting Mammalian Target of Rapamycin Signaling Pathway by Rapamycin on Regulatory Immune T Cell/Regulatory Immune T Cell Immune Homeostasis Imbalance in Heart Failure Mice

作　　者：王清[1] 梁小燕 美迪娜·叶尔肯 芦颜美[1] WANG Qing;LIANG Xiaoyan;Meidina Yeerken;LU Yanmei(Department of Pacing and Electrophysiology,Xinjiang Key Laboratory of Cardiac Electrophysiology and Cardiac Remodeling,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院起搏电生理科、新疆心电生理与心脏重塑重点实验室,乌鲁木齐830054

出　　处：《中国循环杂志》2024年第11期1125-1132,共8页Chinese Circulation Journal

基　　金：新疆维吾尔自治区天山英才-青年拔尖科技人才项目(2022TSYCCX0101)。

摘　　要：目的:探究哺乳动物雷帕霉素靶蛋白(mTOR)的拮抗剂雷帕霉素(RAPA)调节mTOR信号通路对心力衰竭(心衰)小鼠调节性T细胞(Treg)/辅助性T细胞17(Th17)免疫自稳失衡的影响。方法:选用34只健康C57BL/6小鼠,采用冠状动脉左前降支结扎术构建心肌梗死后心衰小鼠模型。按随机数字表法将其随机分为假手术组、心衰组、低剂量RAPA组(RAPAL组)、中剂量RAPA组(RAPAM组)、高剂量RAPA组(RAPAH组),除心梗后心衰造模手术过程中死亡4只,最终每组6只小鼠。假手术组采取同样手术操作但不结扎。采用尾静脉给药的方式,RAPAL组、RAPAM组、RAPAH组的给药剂量分别为1、2、4 mg/(kg·d),其余组尾静脉注射等量生理盐水,持续干预4周。超声心动图检测小鼠心脏结构和功能;苏木素-伊红(HE)染色观察心肌组织病理形态;天狼星红染色观察心肌纤维化程度;流式细胞技术检测外周血Treg、Th17水平;蛋白免疫印迹(Western blot)法检测心肌组织mTOR、磷酸化mTOR(p-mTOR)蛋白表达水平。结果:超声心动图检测发现,与假手术组相比,心衰组左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)均显著降低(P均<0.01),左心室舒张末期容积(LVEDD)、左心室收缩末期容积(LVEDS)均显著升高(P均<0.05);RAPAL、RAPAM、RAPAH组LVEF、LVFS均显著高于心衰组(P均<0.05)。RAPAM、RAPAH组LVEDS均显著低于心衰组(P均<0.05)。HE染色、天狼星红染色发现,与假手术组相比,心衰组小鼠的心肌组织排列紊乱、断裂,心肌纤维化。与心衰组相比,RAPAL组、RAPAM组、RAPAH组小鼠的心肌组织排列紊乱、断裂,心肌纤维化等现象有所改善;流式细胞技术、Western blot法检测发现,与假手术组相比,心衰组小鼠外周血Treg百分比降低,Th17细胞百分比升高(P<0.01),Treg/Th17比值降低(P<0.01)、心肌组织p-mTOR蛋白表达水平差异无统计学意义(P>0.05)。与心衰组相比,RAPAL组、RAPAM组、RAPAH组小鼠的外周血Treg百分Objectives:To explore the effect of mammalian target of rapamycin(mTOR)antagonist rapamycin(RAPA)on the imbalance of regulatory immune T cell(Treg)/regulatory immune T cell(Th17)immune homeostasis in mice with heart failure(HF)by regulating mTOR signaling pathway.Methods:The model of heart failure after myocardial infarction was constructed by ligation of the left anterior descending coronary artery in 34 healthy C57BL/6 mice.The mice were randomly divided into sham operation group,heart failure(HF)group,low-dose(1 mg/[kg·d])RAPAgroup(RAPAL group),medium-dose(2 mg/[kg·d])RAPA group(RAPAM group)group and high-dose(4 mg/[kg·d])RAPA group(RAPAH group)according to the random number table method,with 6 mice in each group.RAPA was administered via tail vein for 4 weeks,and the other groups were injected with the same amount of normal saline by tail vein for 4 weeks.The cardiac structure and function of mice were evaluated by echocardiography.The morphological changes of myocardial tissue were observed by hematoxylin-eosin(HE)staining.The degree of myocardial fibrosis were observed by Sirius red staining The levels of Treg and Th17 cells in peripheral blood were detected by flow cytometry.Western blot was used to detect the protein expression of mTOR,phosphorylated-mTOR(p-mTOR)in myocardial tissue.Results:Echocardiography examination revealed that compared with sham operation group,left ventricular ejection fractions(LVEF)and left ventricular short axis shortening rate(LVFS)were significantly lower(both P<0.01),and Left ventricular end-diastolic Diameter(LVEDD)and Left ventricular end-systolic diameter(LVEDS)were significantly larger(both P<0.05)in HF group.Compared with HF group,LVEF in RAPAL group,RAPAM group and RAPAH group were significantly higher(all P<0.01),LVFS were significantly higher(all P<0.05).LVEDS in RAPAM and RAPAH groups were significantly lower than those in HF group(both P<0.05).HE staining and Sirius red staining showed that compared with the sham operation group,the myocardial tissue of the HF gr

关 键 词：心力衰竭 免疫调节 调节性T细胞/辅助性T细胞17 哺乳动物雷帕霉素靶蛋白 

分 类 号：R54[医药卫生—心血管疾病]