RORα过表达病毒载体尾静脉注射对多柔比星诱导小鼠心脏毒性的抑制作用及其机制  

Effect of RORαoverexpression viral vector on doxorubicin-induced cardiotoxicity in mice and its mechanism

作　　者：赵小华 李艳 张卫泽 徐臣年 刘治国 ZHAO Xiaohua;LI Yan;ZHANG Weize;XU Chennian;LIU Zhiguo(Department of Cardiology,Xi'an International Medical Center Hospital,Xi'an 710100,China;不详)

机构地区：[1]西安国际医学中心医院心脏内科,西安710100 [2]中国人民解放军空军军医大学西京医院心血管外科

出　　处：《山东医药》2024年第23期36-40,共5页Shandong Medical Journal

基　　金：陕西省自然科学基础研究计划项目(2024JC-YBQN-0793)。

摘　　要：目的观察维甲酸相关孤核受体α(RORα)过表达病毒载体尾静脉注射对多柔比星(DOX)诱导小鼠心脏毒性(DIC)的影响,并探讨其可能的机制。方法将80只雄性C57BL/6小鼠随机分为Con组、DIC组、DIC+RORα组、DIC+RORα+EX527组,每组20只。DIC+RORα组和DIC+RORα+EX527组经尾静脉注射过表达RORα基因的腺相关病毒9型(AAV9)载体,DIC组经尾静脉注射AAV9空白对照载体;AAV9载体注射3周后,除Con组外均采用尾静脉注射DOX的方法建立DIC小鼠模型;DIC+RORα+EX527组每次DOX注射后腹腔注射沉默信息调节因子(SIRT1)信号通路特异性阻断剂EX527。各组分组处理后4周,经胸超声心动图检查测算左心室射血分数(LVEF)和左心室缩短分数(LVFS),处死后取眼眶血和心肌组织。采用HE染色观察心肌组织病理情况,ELISA法检测血清肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、肌钙蛋白I(cTnI)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α),TUNEL染色后计算细胞凋亡率,二氢乙锭法检测心肌组织活性氧(ROS)表达,比色法检测心肌组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)表达,免疫荧光染色观察心肌组织巨噬细胞浸润和NF-κB p65表达,Western blotting法检测心肌组织cleaved Caspase-3、RORα、SIRT1蛋白表达并计算Ac-NF-κB p65/NF-κB p65、Ac-Foxo1/Foxo1,实时荧光定量PCR法检测心肌组织RORα、SIRT1 mRNA表达。结果与Con组比较,DIC组小鼠心肌纤维排列紊乱且断裂明显,心肌细胞空泡化显著;与DIC组、DIC+RORα+EX527组比较,DIC+RORα组小鼠心肌纤维断裂及心肌细胞空泡化均减轻。LVEF、LVFS及心肌组织SOD、GSH-Px:Con组>DIC+RORα组>DIC组、DIC+RORα+EX527组(P均<0.05)。血清CK-MB、LDH、cTnI、IL-1β、IL-6、TNF-α及心肌组织细胞凋亡率、cleaved Caspase-3蛋白、ROS、MDA、巨噬细胞浸润、NF-κB p65:Con组<DIC+RORα组<DIC组、DIC+RORα+EX527组(P均<0.05)。RORαmRNA及蛋Objective To investigate the effect of retinoid-related orphan receptorα(RORα)overexpression viral vector on doxorubicin-induced cardiotoxicity(DIC)in mice and to explore its potential mechanism.Methods Eighty male C57BL/6 mice were randomly divided into four groups:Con group,DIC group,DIC+RORαgroup and DIC+RORα+EX527 group,with 20 mice in each group.Mice in the DIC+RORαgroup and DIC+RORα+EX527 group were injected with ade⁃no-associated virus type 9(AAV9)vector that overexpressed RORαthrough the tail vein,while mice in the DIC group were injected with the blank control vector of AAV9.After three weeks of AAV9 vector injection,the DIC mouse model was established by DOX injection through tail vein except for the Con group.DIC+RORα+EX527 group was intraperitone⁃ally injected with SIRT1 specific inhibitor EX527 after each DOX injection.After 4 weeks,left ventricular ejection frac⁃tion(LVEF)and left ventricular fractional shortening(LVFS)were measured using cardiac ultrasonography.Orbital blood and myocardial tissues were taken after death.HE staining was used to observe the pathological changes of myocardial tis⁃sues.Serum creatine kinase-MB(CK-MB),lactate dehydrogenase(LDH),cardiac troponin I(cTnI),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by ELISA.The apoptosis rate was calculat⁃ed after TUNEL staining,the expression of reactive oxygen species(ROS)in myocardial tissue was detected by DHE stain⁃ing,the expression levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in the myocardial tissues were detected by colorimetry method,and the macrophage infiltration and NF-κB p65 expression in the myocardial tissues were observed by immunofluorescence staining.The protein expression levels of cleaved Caspase-3,RORα,and SIRT1 were detected by Western blotting,and Ac-NF-κB p65/NF-κB p65 and Ac-foxo1/foxo1 were calcu⁃lated.The mRNA expression levels of RORαand SIRT1 in the myocardial tissues were detected by real-

关 键 词：维甲酸相关孤核受体α 多柔比星 心脏毒性 氧化应激 炎症反应 沉默信息调节因子1 

分 类 号：R541.9[医药卫生—心血管疾病]