灵芝三萜联合外源性GM1对癫痫大鼠认知功能和海马突触结构的影响  

Effects of ganoderma triterpenoids combined with exogenous GM1 on cognitive function and hippocampal synaptic structure in rats with epilepsy

作　　者：农雪娟 覃丽娜 黄瑜 金佳熹 周冰玉 洪剑威 赵爽[2] Nong Xuejuan;Qin Lina;Huang Yu;Jin Jiaxi;Zhou Bingyu;Hong Jianwei;Zhao Shuang(Department of Laboratory Medicine,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China;Preclinical Medicine College,Youjiang Medical University for Nationalities,Baise 533000,China;Clinical Medicine College,Youjiang Medical University for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院附属医院检验科,百色533000 [2]右江民族医学院基础医学院,百色533000 [3]右江民族医学院临床医学院,百色533000

出　　处：《中华行为医学与脑科学杂志》2022年第10期918-925,共8页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金(81760249);广西中医药管理局科研项目(GZZC2019143);国家级大学生创新创业训练计划项目(201810599002);广西自治区级大学生创新创业训练计划项目(201910599048)。

摘　　要：目的探讨灵芝三萜联合外源性单唾液酸四己糖神经节苷脂(monosialotetrahexosyl ganglioside, GM1)对戊四氮(pentylenetetrazol, PTZ)致痫大鼠认知功能及海马神经元突触超微结构的影响。方法将40只雄性清洁级SD大鼠按照随机数字表法分为空白对照组、癫痫模型组、灵芝三萜组、GM1组、灵芝三萜联合GM1组, 每组8只。采用腹腔注射亚惊厥剂量PTZ( 35 mg·kg^(-1)·d^(-1))的方法建立慢性癫痫大鼠模型, 1次/d, 连续28 d;用药各组大鼠在每日腹腔注射PTZ的同时按分组给予相应药物( GM1:腹腔注射30 mg·kg^(-1)·d^(-1), 灵芝三萜:灌胃1 000 mg·kg^(-1)·d^(-1))。应用Morris水迷宫观测各组大鼠学习记忆能力;透射电镜观察海马神经元突触超微结构;免疫荧光染色法观察大鼠海马CA1区丝切蛋白(cofilin)、突触素(synaptophysin, SYN)的表达, 同时用Western blot检测大鼠海马cofilin及其磷酸化p-cofilin和SYN的蛋白表达。采用SPSS 17.0软件对数据进行统计分析, 组间多样本采用单因素方差分析(one-way ANOVA), 两两比较采用LSD检验。结果 Morris水迷宫结果显示, 各组间的逃避潜伏期、穿越平台次数、目标象限停留时间均差异有统计学意义(F=5.259, 8.240, 5.961, 均P<0.05)。与癫痫模型组相比, 灵芝三萜组、GM1组和灵芝三萜联合GM1组大鼠的逃避潜伏期[(20.31±7.39)s, (21.81±6.05)s, (17.66±4.76)s]均短(均P<0.05), 穿越平台次数[(4.63±1.41)次, (4.50±1.93)次, (5.50±1.77)次]均多(均P<0.05), 目标象限停留时间[(31.91±5.00)s, (30.49±5.72)s, (35.70±5.34)s]均长(均P<0.05), 以灵芝三萜联合GM1组变化最明显(P<0.01)。透射电镜结果显示, 各组海马神经元突触数量、突触间隙宽度、突触后致密物厚度、突触后膜活性区长度均差异有统计学意义(F=3.693, 7.201, 5.012, 4.033, 均P<0.05)。与癫痫模型组比较, 灵芝三萜组、GM1组和灵芝三萜联合GM1组神经元突触数量[(8.00±1.79)个, (7.83±1.84)个,Objective To study the intervention effect of ganoderma triterpenoids combined with exogenous monosialotetrahexosyl ganglioside(GM1)on cognitive dysfunction and synaptic ultrastructure of hippocampal neurons in rats with epilepsy caused by pentylenetetrazol(PTZ).Methods A total of 40 Sprague-Dawley rats were divided into blank control group,epileptic model group,ganoderma triterpenoids group,GM1 group and GM1 combined with ganoderma triterpenoids group according to the random number table method(n=8 in each group).The rats were intraperitoneally injected with PTZ subconvulsant dose(35 mg·kg^(-1)·d^(-1))once a day for 28 days to replicate the models of chronic epilepsy.And the rats in different medication groups were given corresponding administration based on daily intraperitoneal injection of PTZ(GM1:intraperitoneal injection of 30 mg·kg^(-1)·d^(-1),ganoderma triterpenoids:gavage 1000 mg·kg^(-1)·d^(-1)).Morris water maze was used to test the spatial exploration and learning and memory ability of epileptic rats.Transmission electron microscopy was used to observe the ultrastructure of hippocampal neurons in epileptic rats.Immunofluorescence staining was used to observe expression levels of cofilin and SYN protein in hippocampus CA1 of rats.In addition,Western blot was used to detect the expression levels of cofilin,p-cofilin and synaptophysin(SYN)protein in hippocampus of rats.SPSS 17.0 software was used for statistical analysis.Repeated one-way ANOVA was used for comparing among groups,LSD test was used for pairwise comparisons.Results Morris water maze results showed that there were statistically significant differences in escape latency,times of crossing the platform and time spent in the target quadrant among the groups(F=5.259,8.240,5.961,all P<0.05).Compared with the epilepsy model group,the escape latencies((20.31±7.39)s,(21.81±6.05)s,(17.66±4.76)s)of the ganoderma triterpenoids group,GM1 group and GM1 combined with ganoderma triterpenoids group were shorter(all P<0.05),the numbers of crossing the

关 键 词：癫痫 认知功能 突触可塑性 丝切蛋白 灵芝三萜 单唾液酸四己糖神经节苷脂 

分 类 号：R742.1[医药卫生—神经病学与精神病学]