七氟醚可逆性下调糖尿病模型大鼠心肌生物钟蛋白BMAL1的表达  

Sevoflurane reversiblely down-regulates BMAL1 expression of myocardium clock gene of diabetes rat models

作　　者：刘慧[1] 韩冲芳 秦小英 于菁 贺建东 杨文曲 LIU Hui;HAN Chongfang;QIN Xiaoying;YU Jing;HE Jiandong;YANG Wenqu(College of Anesthesiology,Shanxi Medical University,Taiyuan 030001;Department of Anesthesiology,Shanxi Baithune Hospital,Taiyuan 030001,China)

机构地区：[1]山西医科大学麻醉学院,山西太原030001 [2]山西白求恩医院麻醉科,山西太原030001

出　　处：《基础医学与临床》2025年第1期70-75,共6页Basic and Clinical Medicine

基　　金：山西白求恩医院优秀青年启动基金(2019YJ11)。

摘　　要：目的观察七氟醚(SEV)对糖尿病大鼠心肌生物钟基因芳香烃受体核转运样蛋白1(BMALl1)表达的影响,并探讨其变化规律。方法雄性SD大鼠60只,体质量200~250 g,将正常大鼠分为吸氧组(NC)、吸七氟醚组(SEV);常规建立糖尿病模型,建立成功后分为吸氧组(DM)、吸七氟醚组(DM+SEV),吸入时间5 h(n=15)。4组试验动物分别在停止麻醉后0、12和24 h处死,分离心肌组织。用Western blot测定生物钟基因BMAL1与其活化酶泛素特异性肽酶9X(USP9X)的表达;HE染色观察心肌组织病理特征以及免疫荧光共定位观察USP9X与BMAL1之间的相互关系。结果停止麻醉后0、12 h,与DM组比较,DM+SEV组BMAL1、USP9X表达均明显下调(P<0.05);停止麻醉后24 h,与DM组比较,DM+SEV组BMAL1、USP9X表达水平的变化差异无统计学意义。HE染色光镜下显示:DM+SEV组在停止麻醉后0、12 h时心肌组织纤维结构排列发生改变,且在停止麻醉后0 h时这种改变最为显著,但在24 h时心肌组织结构排列整齐。免疫荧光共定位结果显示:USP9X与BMAL1蛋白主要分布于心肌细胞质中,两者存在重叠部分。且受七氟醚影响,在停止麻醉后0、12 h两者重叠部分较少,而在24 h时两者重叠部分较多,接近于DM组。结论七氟醚可逆性改变糖尿病大鼠心肌生物钟基因BMAL1表达,该作用在停止麻醉后12 h仍然存在,而在停止麻醉后24 h该作用明显减弱。Objective To observe the effect of sevoflurane(SEV)on the expression of myocardial biological clock gene aromatic hydrocarbon receptor nuclear transport-like protein 1(BMAL1)in diabetic rats and to explore its changes.Methods Sixty healthy male SD rats with a body mass of 200-250 g were divided into oxygen inhalation group(NC)and sevoflurane inhalation group(SEV).The diabetic model was routinely replicated,and the model was divided into oxygen group(DM)and sevoflurane group(DM+SEV)with an inhalation time of 5 h(n=15).Four groups of experimental animals were executed at 0,12 and 24 h after the anesthesia was stopped and then myocardial tissue was isolated.Western blot was used to determine the expression level of biological clock gene BMAL1 protein and its activation enzyme USP9X;HE staining microspy to observe the pathological changes of myocardial tissue and immuno-fluorescence co-localization to observe the relationship between USP9X and BMAL1.Results At 0 and 12 h after stopping anesthesia,the expression of BMAL1 and USP9X in the DM+SEV group was significantly down-regulated as compared with the DM group,and the expression of BMAL1 and USP9X in the DM+SEV group was significantly down-regulated(P<0.05)at 24 h after stopping anesthesia(P>0.05).HE staining microscopy found changes of myocardial tissue structure in the DM+SEV group at 0 and 12 hrs after stopping anesthesia.This change was most significant at 0 h after stopping anesthesia,but the myocardial tissue structure was neatly arranged at 24 h.The results of immuno-fluorescence colocalization showed that USP9X and BMAL1 proteins were mainly distributed in the cytoplasm of cardio-myocardium with and overlapping parts between them.Under the influence of sevoflurane,there was less overlap between the two at 0 and 12 hrs after stopping anesthesia and more overlap between the two at 24 h,which was close to that of the DM group.Conclusions Sevoflurane reversibly changes the expression of myocardial circadian clock gene BMAL1 in diabetic rats and this change still

关 键 词：七氟醚 糖尿病 心肌 芳香烃受体核转运样蛋白1(BMAL1) 泛素特异性肽酶9X(USP9X) 

分 类 号：R587[医药卫生—内分泌]