基于GABA信号通路和神经-免疫-内分泌网络探讨米泔制僵蚕“粉末入药”的“息风止痉”作用机制  

Xifengzhijing(息风止痉)Mechanism of Rice-Washing Water-Processed Jiangcan(僵蚕)Powder Based on GABA Signaling Pathway and Neuro-Immune-Endocrine Network

作　　者：胡美变 何丽英 王丹[2] 吴纯洁[2] 刘玉杰 Hu Meibian;He Liying;Wang Dan;Wu Chunjie;Liu Yujie(Shanxi University of Chinese Medicine,Jinzhong 030619;Chengdu University of Traditional Chinese Medicine,Chendu 611137)

机构地区：[1]山西中医药大学,晋中030619 [2]成都中医药大学,成都611137

出　　处：《中药药理与临床》2022年第5期118-124,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(编号:82104397);山西省应用基础研究计划项目(编号:20210302124630);山西省高等学校科技创新计划项目(编号:2021L357);山西中医药大学博士科研启动基金项目(编号:2022BK07);山西中医药大学优秀博士毕业生来晋工作奖励经费科研启动基金项目(编号:2022BKS04)。

摘　　要：目的:基于GABA信号通路和神经-免疫-内分泌网络探讨僵蚕“粉末入药”的息风止痉作用及其机制。方法:SD大鼠90只,按随机数字表法分为为正常对照组、模型对照组、地西泮0.6 mg/kg组,米泔制僵蚕水煎液0.5、1和2 g/kg组和米泔制僵蚕粉末0.5、1和2 g/kg组,除正常对照组外,其余组腹腔注射戊四氮(PTZ)建立慢性点燃大鼠模型。正常对照组、模型对照组灌胃生理盐水,阳性药组腹腔注射地西泮0.6 mg/kg,给药组灌胃米泔制僵蚕水煎液或粉末,连续给药30 d。观察大鼠行为学差异和海马组织病理变化;采用ELISA法检测海马组织中神经-免疫-内分泌网络相关分子的表达;采用免疫组化、Western blot和Real-time PCR法检测GABA信号通路相关mRNA及蛋白的表达。结果:与正常对照组相比,模型对照组大鼠癫痫发作潜伏期缩短,持续时间显著延长,海马GABA、5-HT、GRα含量显著降低、Glu、IL-1β、TNF-α含量显著升高(P<0.01),CA1区出现显著病理变化,海马Gabaarα1、Gad 65、Gad 67 mRNA及蛋白表达显著下调,Nmdar1、Gat 1、Gat 3 mRNA及蛋白表达显著上调(P<0.01);相比模型对照组,米泔制僵蚕粉末不同剂量均能显著延长大鼠痫性发作潜伏期,缩短发作时间(P<0.05或P<0.01),升高大鼠海马GABA、5-HT、GRα含量、降低Glu、IL-1β、TNF-α含量(P<0.05或P<0.01),且粉末能明显改善模型大鼠海马CA1区的病理状态,明显调节GABA信号通路相关mRNA和蛋白(GABAA-Rα1、GAD 65、GAD 67、GAT 1和GAT 3)及NMDAR1受体的表达。结论:米泔制僵蚕粉末息风止痉作用机制与调节大鼠海马中GABA信号通路相关mRNA和蛋白的表达有关,并调控IL-1β、TNF-α和GRα的分泌,使GABA、Glu水平及神经-免疫-内分泌网络趋于平衡。Objective:To explore the Xifengzhijing(息风止痉)effect and mechanism of rice-washing water-processed Jiangcan(僵蚕)(RWJC)powder based onγ-aminobutyric acid(GABA)signaling pathway and neuro-immune-endocrine network.Methods:A total of 90 SD rats were randomized into normal control group(10 mL/kg normal saline,ig),model control group(10 mL/kg normal saline,ig),diazepam(DZP,0.6 mg/kg)group(ip),0.5,1,and 2 g/kg RWJC decoction groups(ig),and 0.5,1,and 2 g/kg RWJC powder groups(ig).Pentylenetetrazole(PTZ,ip)was used to induce chronic epilepsy in rats except for the normal control group.The administration lasted 30 days for all the groups.The behaviors of rats and pathological changes of rat hippocampus were observed.The expression of neuro-immune-endocrine network-related molecules in hippocampus was detected by enzyme-linked immunosorbent assay(ELISA).The levels of GABA signaling pathway-related proteins and mRNA were measured by immunohistochemistry,Western blotting,and real-time PCR.Results:Compared with the normal control group,model control group showed short incubation time of seizure,long duration,high content of GABA,serotonin(5-HT),and glucocorticoid receptorα(GRα)in hippocampus,low content of glutamic acid(Glu),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)(P<0.01),obvious pathological changes in CA1 region,low mRNA and protein expression of alpha 1 subunit of the gamma-aminobutyric acid-A receptor(GABAARα1),glutamate decarboxylase(GAD)65,and GAD 67,and high mRNA and protein expression of N-methyl-D-aspartate receptor-1(NMDAR1),GABA transporter type(GAT)-1,and GAT-3(P<0.01).Compared with the model control group,different doses of RWJC powder prolonged incubation time and shortened the duration of rat seizure(P<0.05 or P<0.01),raised the content of GABA,5-HT,and GRα,and decreased the content of Glu,IL-1β,and TNF-αin hippocampus(P<0.05 or P<0.01).RWJC powder significantly alleviated the pathological state of CA1 region in hippocampus of model rat and significantly regulated GABA signal

关 键 词：僵蚕 粉末入药 米泔制 息风止痉 脑内神经递质 炎症因子 

分 类 号：R285[医药卫生—中药学]