基于JAK2/STAT3信号通路探讨温阳解郁方调节小鼠海马小胶质细胞激活的机制  

Mechanism of Wenyang Jieyu Prescription in Regulating Activation of Mouse Hippocampal Microglia Based on JAK2/STAT3 Signaling Pathway

作　　者：王英 巩子汉 梁文青 杨婧雯 岳广欣[1] WANG Ying;GONG Zihan;LIANG Wenqing;YANG Jingwen;YUE Guangxin(Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;Ningxia Chinese Medicine Research Center,Ningxia 750000,China;Hengshanqiao People's Hospital in Changzhou Economic Development Zone,Changzhou 213161,China;Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中医基础理论研究所,北京100700 [2]宁夏回族自治区中医医院暨中医研究院,宁夏750000 [3]江苏常州经济开发区横山桥人民医院,江苏常州213161 [4]中国中医科学院中医临床基础医学研究所,北京100700

出　　处：《中国实验方剂学杂志》2025年第8期88-96,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82174251);北京市自然科学基金面上项目(7232300);中国中医科学院科技创新工程重大攻关项目(CI2021A00607);中国中医科学院科技创新工程项目中医理论传承与创新专项(KYG-202406);中央级公益性科研院所基本科研业务费专项(YZX-202242)。

摘　　要：目的:探讨温阳解郁方调节母婴分离结合二次打击小鼠抑郁样行为的作用机制。方法:仔鼠出生后随机分为空白(NC)组、母婴分离(MS)组、束缚应激(RS)组、母婴分离结合束缚应激(MRS)组、温阳(Wenying)组、解郁(Jieyu)组、温阳解郁(XSF)组、米诺环素(minocycline)组,于第5天(PD5)进行母婴分离,PD21离乳并预防性给药,Wenyang组、Xiaoyao组、XSF组及minocycline组小鼠给药剂量分别为5.85、12.03、16.71 g·kg^(-1)和50 mg·kg^(-1),PD90进行束缚应激。通过糖水、社会交互、旷场、O迷宫行为学实验及高效液相法检测5-羟色胺、多巴胺等神经递质水平进行模型评价;免疫组化法检测海马小胶质细胞标志物离子钙结合衔接分子-1(Iba-1)蛋白表达水平;全自动蛋白表达分析系统检测各组小鼠海马Janus激酶2(JAK2)、信号转导及转录激活因子3(STAT3)蛋白表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测各组小鼠海马小胶质细胞M1型标志物、JAK2/STAT3通路和细胞因子各项指标mRNA表达水平。结果:与NC组比较,MRS组小鼠各行为学表现出抑郁样行为,海马各神经递质水平均明显下降(P<0.05,P<0.01),Iba-1表达增多(P<0.01),JAK2、STAT3蛋白表达水平上升(P<0.05),CD68、CD11b、IL^(-1)β、JAK2、STAT3 mRNA表达显著升高(P<0.01),IL^(-1)0 mRNA表达显著降低(P<0.01)。与MRS组比较,XSF组和minocycline组抑郁样行为有一定的改善,XSF组和minocycline组小鼠海马神经递质含量明显升高(P<0.05,P<0.01),Iba-1表达显著减少(P<0.01),XSF组JAK2、STAT3蛋白表达水平有下降趋势,XSF组和minocycline组小鼠海马CD68、CD11b、JAK2、STAT3及IL^(-1)β mRNA表达明显降低(P<0.05,P<0.01),IL^(-1)0 mRNA表达均明显升高(P<0.05,P<0.01)。结论:温阳解郁方可通过抑制海马小胶质细胞向M1型极化调节母婴分离二次打击小鼠抑郁样行为,且温阳解郁方调节海马小胶质细胞极化可能与JAK2/STAT3通路有关。Objective:To explore the mechanism of the Wenyang Jieyu prescription in regulating depression-like behavior in mice after maternal-infant separation combined with secondary stress.Methods:After birth,the rats were randomly divided into blank(NC)group,maternal-infant separation(MS)group,restraint stress(RS)group,maternal-infant separation combined with restraint stress(MRS)group,Wenyang group,Jieyu group,Wenyang Jieyu(XSF)group,and minocycline group.Maternal infant separation was performed on day 5(PD5),followed by weaning at PD21 and prophylactic administration.The dose of Wenyang group,Xiaoyao group,XSF group and minocycline group were 5.85,12.03,16.71 g·kg^(-1) and 50 mg·kg^(-1),respectively.Restraint stress was applied on PD90.The model was evaluated using glucose,social interaction,open field,and O-maze behavior tests,as well as high-performance liquid chromatography to measure serotonin,dopamine,and other neurotransmitters.The expression level of ionized calcium-binding adaptor molecule-1(Iba-1)protein,a marker of hippocampal microglia,was detected by immunohistochemistry.Protein expression levels of Janus kinase 2(JAK2)and signal transducer and activator of transcription 3(STAT3)in the hippocampus were analyzed by an automatic protein expression analysis system.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect mRNA expression levels of M1 markers,JAK2/STAT3 pathway-related genes,and cytokines in hippocampal microglia in each group.Results:Compared with the NC group,the MRS group exhibited depression-like behavior,with significantly decreased levels of neurotransmitters in the hippocampus(P<0.05,P<0.01),increased expression of Iba-1(P<0.01),and elevated protein levels of JAK2 and STAT3(P<0.05).The mRNA expression levels of CD68,CD11b,IL^(-1)β,JAK2,and STAT3 were significantly increased(P<0.01),while IL^(-1)0 mRNA expression was significantly decreased(P<0.01).Compared with the MRS group,the XSF and minocycline groups showed some improvement in depression-like b

关 键 词：母婴分离 抑郁样行为 小胶质细胞 Janus激酶2(JAK2)/信号转导及转录激活因子3(STAT3)通路 温阳解郁方 

分 类 号：R259[医药卫生—中西医结合] R289[医药卫生—中医内科学] R742[医药卫生—中医学]