转录因子12过表达对脓毒症相关性脑病大鼠大脑皮质的保护作用及其机制  

The protective effect and mechanism of overexpression of transcription factor 12 on the cerebral cortex of rats with sepsis-associated encephalopathy

作　　者：张晓燕[1] 肖东琼[1] 高沪[1] 陈琳[1] 唐发娟 李熙鸿[1] Zhang Xiaoyan;Xiao Dongqiong;Gao Hu;Chen Lin;Tang Fajuan;Li Xihong(Department of Emergency Medicine,Key Laboratory of Birth Defects and Related Diseases of Women and Children(Sichuan University),Ministry of Education,West China Second University Hospital,Sichuan University,Chengdu 610041,Sichuan Province,China)

机构地区：[1]四川大学华西第二医院急诊医学科、出生缺陷与相关妇儿疾病教育部重点实验室,成都610041

出　　处：《中华妇幼临床医学杂志（电子版）》2023年第5期540-549,共10页Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)

基　　金：国家自然科学基金项目(82071353);四川省科技厅重点研发项目(2021YFS0029、2023YFS0025)。

摘　　要：目的探讨转录因子12(Tcf12)过表达,对脓毒症相关性脑病(SAE)大鼠大脑皮质的保护作用及其机制。方法选择144只30 d龄Sprague Dawley(SD)雄性大鼠为研究对象。按照处理方法,将其分为盲肠结扎穿孔术(CLP)组、Tcf12阴性对照组(Veh+CLP组)和Tcf12过表达组(AD-Tcf12+CLP组)和假手术组(Sham组)4组。对Sham组和CLP组大鼠,在其双侧侧脑室各注射5μL生理盐水;对AD-Tcf12+CLP组大鼠采取硬脑膜微量注射5μL AD-Tcf12,Veh+CLP组,则硬脑膜微量注射5μL Veh。侧脑室注射后48 h,对CLP组、Veh+CLP组和AD-Tcf12+CLP组实施CLP,对Sham组仅开腹后翻看盲肠。在造模后3、6、12、24、48 h监测4组大鼠平均动脉压(MAP)、心率和神经行为学评分;其中24 h时,采取苏木精-伊红(HE)染色,激光共聚焦扫描显微镜观察大鼠大脑皮质病理改变;采用Western blotting检测大鼠大脑皮质蛋白(Tcf12、cleaved caspase-3、Bax、Bcl2、P38、p-P38)表达;免疫荧光染色观察在大鼠大脑皮质神经细胞中Tcf12相对表达水平。本实验获得四川大学华西第二医院动物保护研究委员会批准[批准编号:2020动伦审批第(申022)号],并且按照相应的指南和规则进行动物实验。结果①与Sham组比较,CLP组和Veh+CLP组大鼠MAP、神经行为学评分在造模后不同时间点(3、6、12、24、48 h)呈下降趋势,心率呈增高趋势,均于24 h时最显著,并且差异均有统计学意义(P<0.05)。②与Veh+CLP组比较,AD-Tcf12+CLP组MAP、神经行为学评分增高,心率下降,于造模后24 h时最明显,并且差异有统计学意义(P<0.05)。②Western blotting检测结果显示,与Sham组相比,CLP组造模后12、24、48 h时,Tcf12相对表达水平比较,差异均有统计学意义(P=0.045、0.019、0.030),其中24 h时达低谷。③造模后24 h时,免疫荧光染色显示,Tcf12表达主要定位在神经元细胞质中。其中24 h时,CLP组SD大鼠Tcf12相对荧光信号为75.11±6.39,显著低于Sham组的101.79±3.89,并且差异有�Objective To investigate protective effects and mechanism of overexpressed transcription factor 12(Tcf12)on the cortex of rats with sepsis-associated encephalopathy(SAE).Methods A total of 144 male Sprague Dawley(SD)rats at 30 days of age were selected into this study.They were randomly divided into four groups according to the processing method:cecal ligation and perforation(CLP)model group(CLP group),Tcf12 negative control group(Veh+CLP group),Tcf12 overexpression group(AD-Tcf12+CLP group),and sham surgery group(Sham group).The Sham group and CLP group received 5μL physiological saline injected into each lateral ventricle.The AD-Tcf12+CLP group received 5μL AD-Tcf12,and the Veh+CLP group received 5μL Veh.After 48 hours of lateral ventricle injection,CLP surgery was performed.The Sham group only underwent laparotomy and examination of the cecum.The mean arterial pressure(MAP),heart rate,and neurobehavioral scores were monitored at 3,6,12,24,and 48 hours after modeling.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in the cerebral cortex of each group at 24 hours after modeling.Western blotting was used to detect the protein expression of Tcf12,cleaved caspase-3,Bax,Bcl2,P38,and p-P38 in the cerebral cortex of rats.Immunofluorescence staining was used to observe the expression of Tcf12 in neurons of the cerebral cortex.This experiment was approved by the Animal Protection Research Committee of West China Second University Hospital,Sichuan University[Approval No.2020(022)],and animal experiments were carried out in accordance with corresponding guidelines and regulations.Results①Compared with Sham group,the MAP and neurobehavioral scores of CLP group and Veh+CLP group showed a decreasing trend at different time points,while heart rate showed an increasing trend.The most significant changes occurred at 24 hours after modeling,and the differences were statistically significant(P<0.05).In addition,the neurobehavioral scores of CLP group and Veh+CLP group were less than 6 points at 24

关 键 词：转录因子12 细胞凋亡 脓毒症相关性脑病 神经元 作用机制 大鼠 

分 类 号：R459.7[医药卫生—急诊医学] R741[医药卫生—治疗学]