癫痫清颗粒介导线粒体自噬对P301S小鼠tau蛋白的影响  

Effects of Dianxianqing granules on tau protein in P301S mice by regulating mitophagy

作　　者：齐越 贾冬 张艳军 谢奇 陈桂娟 谭东明 陈二华 丁旭 殷紫 QI Yue;JIA Dong;ZHANG Yanjun;XIE Qi;CHEN Guijuan;TAN Dongming;CHEN Erhua;DING Xu;YIN Zi(Dept.of Pharmaceutical Technology,Xuzhou Pharmaceutical Vocational College of Jiangsu Province,Jiangsu Xuzhou 221116,China;Dept.of Pharmaceutical Technology,the Second Affiliated Hospital ofLiaoning University of Traditional Chinese Medicine,Shenyang 110034,China;College of Pharmacy andTraditional Chinese Medicine,Jiangsu Nursing Vocational College,Jiangsu Huaian 223200,China)

机构地区：[1]江苏省徐州医药高等职业学校药学技术系,江苏徐州221116 [2]辽宁中医药大学附属第二医院药理室,沈阳110034 [3]江苏护理职业学院药学与中药学院,江苏淮安223200

出　　处：《中国药房》2023年第14期1712-1718,共7页China Pharmacy

基　　金：徐州市科技项目(No.KC22134)。

摘　　要：目的研究癫痫清颗粒介导线粒体自噬对P301S小鼠tau蛋白的影响。方法将36只P301S小鼠按体重分为模型组、癫痫清颗粒组(12.48 g/kg)和盐酸多奈哌齐组(阳性对照,1.3 mg/kg),每组12只;另取C57BL6小鼠10只,作为对照组。各药物组小鼠灌胃相应药液,对照组和模型组灌胃等体积水;灌胃体积均为20 mL/kg,每天1次,连续5个月。实验期间,观察各组小鼠的一般状况;末次给药后,借助Y迷宫和Morris水迷宫实验检测小鼠的学习记忆能力;以苏木精-伊红染色法观察其脑组织形态学改变,以尼氏染色法观察其脑组织神经细胞结构和尼氏体数量;以免疫组织化学法检测其脑组织中丝氨酸202/苏氨酸205位点磷酸化tau(简称“AT8”)蛋白的表达水平,Western blot法检测脑组织中线粒体自噬相关蛋白[PTEN诱导激酶1(PINK1)、Parkin、微管相关蛋白1轻链3B(LC3B)、p62]、突触相关蛋白[突触后密度蛋白95(PSD-95)、突触生长蛋白(SYP)、生长相关蛋白43(GAP-43)]的表达水平和tau蛋白的磷酸化水平[以丝氨酸199(Ser199)、Ser202的磷酸化水平表示]。结果模型组小鼠出现毛发变白、体重下降、下肢瘫痪等症状;其脑组织海马结构欠完整,神经细胞膜及细胞结构亦不完整;其自发交替反应率、穿越平台次数、细胞质内尼氏体数量和PINK1、Parkin、LC3B、SYP、GAP-43、PSD-95蛋白的表达水平均较对照组显著降低,游泳潜伏期(第4、5天)、AT8和p62蛋白的表达水平、Ser199和Ser202蛋白的磷酸化水平均较对照组显著升高或延长(P<0.05或P<0.01)。与模型组比较,各药物组小鼠上述症状和指标均得以显著改善(P<0.05或P<0.01)。结论癫痫清颗粒可有效改善P301S小鼠的认知功能障碍,作用可能与介导线粒体自噬,减少tau蛋白过度磷酸化,上调脑组织中突触相关蛋白的表达,修复受损神经细胞有关。OBJECTIVE To study the effects of Dianxianqing granules on the tau protein in P301S mice by regulating mitophagy.METHODS Totally 36 P301S mice were randomly divided into model group,Dianxianqing granule group(12.48 g/kg),donepezil hydrochloride group(positive control,1.3 mg/kg),with 12 mice in each group;another 10 C57BL6 mice were selected as control group.Administration groups were given relevant drug solutions intragastrically,and control group and model group were given constant volume of water intragastrically.The gavage volume was 20 mL/kg,once a day,for consecutive 5 months.During the experiment,the general condition of mice was observed in each group.After the last medication,the learning and memory ability was determined by Y maze test and Morris water maze test;HE staining was used to observe the morphological changes in brain tissue,and Nissl staining was used to observe the structure of neural cells and the number of Nissl bodies in cerebral tissue.Immunohistochemistry was used to detect the expressions of phospho-tau serine 202/threonine 205(abbreviated as AT8)in brain tissue.Western blot assay was used to determine the expressions of mitophagy-associated proteins[PTEN-induced putative kinase-1(PINK1),Parkin,microtubule-associated protein 1 light chain 3B(LC3B),p62],synaptic-associated proteins[postsynaptic density protein-95(PSD-95),synaptophysin(SYP),and growth-associated protein-43(GAP-43)]and the phosphorylation of tau protein[expressed by the phosphorylation levels of serine 199(Ser199)and Ser202]in brain tissue.RESULTS The mice in model group showed symptoms such as white hair,decreased body mass,and lower limb paralysis,with incomplete hippocampal structures in their brain tissue,as well as incomplete cell membrane edges and cell structures;the spontaneous alternating response rate,the times of crossing platform,the number of Nissl bodies,the protein expressions of PINK1,Parkin,LC3B,SYP,GAP-43,and PSD-95 were decreased significantly,compared with control group;swimming latency(fourth and fifth

关 键 词：癫痫清颗粒 阿尔茨海默病 线粒体自噬 TAU蛋白 突触 神经细胞 学习记忆障碍 

分 类 号：R965[医药卫生—药理学]