摘要
目的研究定痫丸对急性期癫痫小鼠海马组织Wnt/β-Catenin信号通路相关蛋白及神经元凋亡的影响,探讨其可能的疗效机制。方法取100只8~10周龄SPF级雄性小鼠,其中20只用于模型失败备用小鼠,将80只小鼠随机分为4组,分别为模型组(A)、生理盐水组(B)、卡马西平组(C)、定痫丸组(D),每组20只,采用腹腔注射氯化锂-匹罗卡品的方法制作急性期癫痫小鼠模型,A组不予干预,B组实验全程均给予生理盐水,C组予卡马西平混悬液灌胃,D组予定痫丸混悬液灌胃,应用免疫组化和Western Blot分别检测海马组织内β-Catenin(β-连环蛋白),Cyclin D1(细胞周期蛋白D1)和Wnt3a的蛋白表达水平,运用Tunel法检测神经元的凋亡情况。结果与生理盐水组比较,模型组海马组织中的β-Catenin、Wnt3a的蛋白表达明显上调(P<0.05),在海马CA1区和CA3区的凋亡阳性细胞增多(P<0.05);与模型组比较,定痫丸组中的β-Catenin、Wnt3a的蛋白表达下调(P<0.05),在海马CA1区和CA3区的凋亡阳性细胞减少(P<0.05);在4组海马组织中均未检测到Cyclin D1蛋白的表达。结论定痫丸可能是通过下调Wnt/β-Catenin信号通路上的β-Catenin、Wnt3a的蛋白的表达,抑制了海马神经元的凋亡,达到治疗癫痫的作用。
Objective To research the effect of Dingxianwan on Wnt/β-Catenin signaling pathway-related protein and its neuronal apoptosis in hippocampus of mice with acute epilepsy,explore possible mechanisms of efficacy.Method Take 100 male mice of 8~10 weeks old SPF grade,20 of them were used when the modeling failed.80 mice were randomly divided into 4 groups,Model group(A), saline group(B),carbamazepine group(C),Dingxianwan group(D), 20 in each group,preparation of acute epilepsy mouse model by intraperitoneal injection of lithium chloride-pilocarpine,Group A did not give interventions,Group B was given normal saline throughout the experiment.Group C was ig administered with carbamazepine suspension,Group D was ig administered Dingxianwan suspension.The expression levels of β-Catenin(β-Catenin),Cyclin D1(cyclin D1) and Wnt3 a in hippocampus were detected by immunohistochemistry and western blot,Detection of neuronal apoptosis using Tunel method.Result Compared with saline group,the protein expression levels of β-Catenin and Wnt3 a in the hippocampus of the model group were significantly up-regulated(P<0.05),increaseed apoptosis-positive cells in the CA1 and CA3 regions of hippocampus(P<0.05);compared with the model group,the protein expression levels of β-Catenin and Wnt3 a in the Dingxianwan group were down-regulated(P<0.05),apoptosis-positive cells in the hippocampal CA1 and CA3 regions were decreased(P<0.05);and the expression of Cyclin D1 protein was not detected in the hippocampus of the four groups.Conclusion Dingxianwan may down-regulate the expression of β-Catenin and Wnt3 a proteins in the Wnt/β-Catenin signaling pathway,inhibition of hippocampal neuronal apoptosis,Achieve the purpose of treating epilepsy.
作者
刁丽梅
李华琼
张庆梅
卢玲
李欢
廖现秋
吴琼
陈爱玲
杨东
仇小强
DIAO Limei;LI Huaqiong;ZHANG Qingmei;LU Ling;LI Huan;LIAO Xianqiu;WU Qiong;CHEN Ailing;YANG Dong;QIU Xiaoqiang(Guangxi Medical University,Nanning 530021.Guangxi,China;The First Affiliated Hospital of Guangxi Universtiy of Chinese Medicine,Namiing 530023,Guangxi,China;Guangxi University of Chinese Medicine,Nanning 530000,Guangxi,China)
出处
《中华中医药学刊》
CAS
北大核心
2020年第3期45-49,I0014,共6页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金(81760809)
广西自然科学基金(2017GXNSFAA198294)
广西医疗卫生适宜技术开发与推广应用项目(S2017049)。
