摘要
目的探讨黏附分子nectin-1的表达对大鼠癫痫发作及苔藓纤维出芽的影响。方法(1)选取成年雄性SD大鼠36只按照随机数字表法分为对照组(n=6)、空载体组(n=6)和病毒干扰组(n=24),其中病毒干扰组按病毒注射后时间点进一步分为3 d、7 d、14 d及30 d 4个亚组,各亚组6只。采用Western blotting实验检测各组大鼠海马区nectin-1蛋白表达情况。(2)另选取成年雄性SD大鼠12只按照随机数字表法分为病毒干扰癫痫组(n=6)及空载体癫痫组(n=6)。向2组大鼠海马分别注入等量基因干扰慢病毒和慢病毒空载体并点燃匹罗卡品癫痫模型,观察2组大鼠行为学变化,通过Timm染色观察大鼠海马区苔藓纤维出芽的情况。结果(1)各组大鼠海马区nectin-1蛋白表达差异有统计学意义(F=76.120,P=0.000)。与对照组及空载体组比较,病毒干扰组中7 d、14 d、30 d亚组大鼠海马组织中nectin-1表达均明显降低,差异有统计学意义(P<0.05)。(2)大鼠行为学比较:病毒干扰癫痫组大鼠点燃所需时间[(34.33±2.38)min]较空载体癫痫组大鼠[(24.50±2.06)min]明显延长,差异有统计学意义(t=7.650,P=0.000)。发作级别:病毒干扰癫痫组大鼠造模后1 h内各时间点癫痫发作级别均较空载体癫痫组大鼠低,差异具有统计学意义(P<0.05)。大鼠癫痫点燃后至30 d,与空载体癫痫组比较,病毒干扰癫痫组大鼠出现自发发作时间延长,自发发作次数减少,差异均具有统计学意义(P<0.05)。Timm染色:病毒干扰癫痫组Timm染色评分[(3.500±0.224)分]较空载体癫痫组大鼠[(4.667±0.211)分]明显降低,差异具有统计学意义(t=9.289,P=0.000)。结论癫痫大鼠海马区nectin-1表达被抑制后能延缓大鼠癫痫发生,减少癫痫的自发发作次数,其作用机制可能与nectin-1减少海马区异常神经回路苔藓纤维出芽的形成有关。
Objective To explore the effect of adhesion molecule nectin-1 expression on epileptic seizure and mossy fiber sprouting.Methods(1)Adult male SD rats were randomly divided into control group(n=6),empty vector group(n=6)and lentivirus interfered group(n=24),and rats in the lentivirus interfered group were further divided into 4 subgroups(n=6)according to the time points of lentivirus injection(3,7,14 and 30 d after injection).The protein expression of nectin-1 in the hippocampus of rats was detected by Western blotting.(2)Another 12 male SD rats were randomly selected and divided into lentivirus epilepsy group(n=6)and empty vector epilepsy group(n=6).Lentivirus and empty lentivirus vector were injected into the hippocampus of the above two groups respectively,and then,pilocarpine epilepsy models were kindled;the behavior changes of these rats were recorded,and changes of mossy fiber sprouting in the hippocampus were observed by Timm staining.Results(1)There were significant differences in hippocampal nectin-1 protein expressions in each group(F=76.120,P=0.000);the nectin-1 expression in the 7,14,and 30 d subgroups was significantly decreased as compared with that in the control group and empty vector group(P<0.05).(2)The results of behavior changes showed that the times required for the lentivirus epilepsy group to be kindled successfully([34.33±2.38]min)were significantly longer than those for the empty vector epilepsy group([24.50±2.06]min,t=7.650,P=0.000).In terms of seizure level,the seizure level of rats in the lentivirus epilepsy group was significantly lower at each time point within one h modeling than that of rats in the empty vector epilepsy group(P<0.05).After kindling,the time of spontaneous seizure appeared in the lentivirus epilepsy group was significantly longer than that in the empty vector epilepsy group,and the frequencies of spontaneous seizure in the lentivirus epilepsy group were significantly decreased as compared with those in the empty vector epilepsy group(P<0.05).Timm staining scores in
作者
吴小芳
于云莉
冯占辉
李小平
黄书蕾
Wu Xiaofang;Yu Yunli;Feng Zhanhui;Li Xiaoping;Huang Shulei(Department of Neurology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of General Medicine,Jingzhou Central Hospital,Jingzhou 434000,China)
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2020年第10期983-987,共5页
Chinese Journal of Neuromedicine
基金
贵州省科技计划项目(黔科合LH字[2016]7238号)。
