脑创伤后Reg3g的表达及对早期癫痫发作的作用研究  

Expression of Reg3g after brain trauma and its effect on early epilepsy in mice

作　　者：周天钧 程赛宇 ZHOU Tianjun;CHENG Saiyu(Center of Neurological Diseases,the Third Affiliated Hospital of Chongqing Medical University,Chongqing,401120,China)

机构地区：[1]重庆医科大学附属第三医院神经疾病中心,重庆401120

出　　处：《陆军军医大学学报》2023年第5期386-392,共7页Journal of Army Medical University

基　　金：国家自然科学基金面上项目(81671190)。

摘　　要：目的探讨Reg3g信号激活在创伤性脑损伤(traumatic brain injury,TBI)脑组织中的时空表达规律和对TBI早期癫痫发作敏感性的影响。方法采用控制性皮质撞击(controlled cortical impact,CCI)制备TBI模型,在分子与形态学实验中,60只C57BL/6N野生型(wild type,WT)小鼠按照随机数字表法分为4组:sham组、CCI 24 h组、CCI 72 h组、CCI 1周组。通过RT-PCR、Western blot、免疫荧光等方法检测Reg3g的时空表达规律。采用海人酸(kainic acid,KA)诱导癫痫,在脑创伤癫痫发作实验中,WT小鼠和Reg3g KO小鼠各30只,按照随机数字表法分组,每组15只,分别为WT(CCI)+NS组、Reg3g KO(CCI)+NS组、WT(CCI)+KA组、Reg3g KO(CCI)+KA组。在CCI基础上进行癫痫诱导发作,并记录在体脑电结果。结果①与sham组相比,CCI损伤侧皮层24 h、72 h、1周后,Reg3g基因和蛋白表达明显升高,差异有统计学意义(P<0.01);②免疫荧光结果显示,在CCI小鼠皮层组织,Reg3g与NeuN-阳性神经元完全共表达;③在生理盐水注射的WT和Reg3g基因敲除小鼠,并没有观察到明显的癫痫波。而在KA注射的WT小鼠和Reg3g基因敲除小鼠,可观察到明显的癫痫波。与WT小鼠相比,KA注射的Reg3g基因敲除小鼠潜伏期明显缩短,总发作时间明显延长,差异有统计学意义(P<0.05)。结论Reg3g对TBI后早期癫痫发作起着明显的保护作用,Reg3g可能是抑制TBI早期癫痫发作的重要调控靶点。ObjectiveTo investigate the temporal and spatial expression profile of Reg3g signal activation in traumatic brain injury(TBI)and its effect on early seizure sensitivity of TBI.MethodsTBI model was established with controlled cortical impact(CCI).In molecular and morphological experiments,60 C57BL/6N wild type(WT)mice were randomly divided into sham group,CCI 24-h group,CCI 72-h group and CCI 1-week group.The temporal and spatial expression of Reg3g was detected by RT-PCR,Western blotting and immunofluorescence assay.Epilepsy was induced by kainic acid(KA).In the brain trauma seizure experiment,30 WT mice and 30 Reg3g KO mice were randomly divided into WT(CCI)+(normal saline,NS)group,Reg3g KO(CCI)+NS group,WT(CCI)+KA group and Reg3g KO(CCI)+KA group.Epileptic seizures were induced on the basis of CCI,and electroencephalography(EEG)was performed in vivo.Results①Compared with the sham group,the mRNA and protein expression of Reg3g in the cortex of the CCI injured side was significantly increased in 24 and 72 h and 1 week after CCI(P<0.01).②The results of immunofluorescence assay showed that Reg3g and NeuN were completely co-expressed in the neurons in the cortex of CCI mice.③No obvious epileptic waves were observed in WT and Reg3g KO mice injected with NS.Significant epileptic waves were observed in WT mice and Reg3g KO mice injected with KA.Compared with the WT mice,the latency of Reg3g KO mice injected with KA was significantly shorter and the total attack time was significantly longer(P<0.05).ConclusionReg3g plays a significant protective role in early seizures after TBI,and it may be an important regulatory target to inhibit early seizures after TBI.

关 键 词：脑创伤 Reg3g 癫痫发作 敏感性 

分 类 号：R651.15[医药卫生—外科学] R741.02[医药卫生—临床医学] R742.1