基于JAK2/STAT3通路介导的星形胶质细胞A1/A2表型转化探讨桃红四物汤对脑缺血再灌注损伤的保护作用  

Protective Effect of Taohong Siwutang on Cerebral Ischemia-reperfusion Injury Based on A1/A2 Phenotype Transformation of Astrocytes Mediated by JAK2/STAT3 Pathway

作　　者：王慧芳 陈新茹 陈梦圆 周娴 韩岚 陈卫东 季兆洁 WANG Huifang;CHEN Xinru;CHEN Mengyuan;ZHOU Xian;HAN Lan;CHEN Weidong;JI Zhaojie(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;Anhui Province Key Laboratory of Chinese Medicinal Formula,Hefei 230012,China;MOE-Anhui Joint Collaborative Innovation Center for Quality Improvement of Anhui Genuine Chinese Medicinal Materials,Hefei 230012,China)

机构地区：[1]安徽中医药大学药学院,合肥230012 [2]安徽省中药复方重点实验室,合肥230012 [3]省部共建安徽道地中药材品质提升协同创新中心,合肥230012

出　　处：《中国实验方剂学杂志》2025年第7期25-34,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82304912,82074152);安徽省高校自然科学研究项目(2023AH050744)。

摘　　要：目的:探讨桃红四物汤对脑缺血再灌注大鼠的影响是否与调节星型胶质细胞极化有关及相关机制研究。方法:将84只雄性SD大鼠随机平均分为假手术组,模型组,桃红四物汤低、中、高剂量组,磷酸川芎嗪片组和AG490组。除假手术组外均进行MCAO/R造模,给药7 d。采用Longa评分法进行神经功能缺损评分,2,3,5-氯化三苯基四氮唑(TTC)染色评估脑梗死体积,实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测皮质补体成分3(C3)、S100钙结合蛋白A10(S100A10)、非受体型酪氨酸蛋白激酶2(JAK2)、信号转导及转录激活蛋白3(STAT3)相关mRNA和蛋白表达差异,血管内皮生长因子-A(VEGF-A)蛋白表达差异,炎症因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)mRNA表达水平;免疫荧光双染检测GFAP和C3、S100A10共定位情况;酶联免疫吸附测定法(ELISA)检测VEGF表达情况。结果:与假手术组比较,模型组脑梗死体积、神经功能损伤显著升高(P<0.01),蛋白C3上升而S100A10降低,信号通路相关蛋白明显升高(P<0.05,P<0.01),VEGF-A蛋白显著降低(P<0.01),相关炎症因子mRNA表达显著升高(P<0.01);共定位荧光强度GFAP和C3显著上升(P<0.01),GFAP和S100A10显著下降(P<0.01),VEGF含量显著升高(P<0.01)。与模型组比较,桃红四物汤中、高剂量和磷酸川芎嗪片组脑梗死体积、神经功能损伤显著降低(P<0.01);桃红四物汤低、中、高剂量,磷酸川芎嗪片组蛋白表达C3降低而S100A10显著升高(P<0.01);桃红四物汤高剂量组,AG490组蛋白及mRNA表达C3、通路相关明显降低(P<0.05,P<0.01),S100A10显著升高(P<0.01),VEGF-A蛋白显著上升(P<0.01),相关炎症因子mRNA表达显著降低(P<0.01);共定位荧光强度GFAP和C3显著降低(P<0.01),GFAP和S100A10显著升高(P<0.01);VEGF含量显著上升(P<0.01)。结论:桃红四物汤发挥对脑缺血再灌注大鼠脑保护作用,其机制与下调JAK2/STATObjective:To investigate whether the effect of Taohong Siwutang on cerebral ischemia-reperfusion(CIRI)injury in rats is related to the regulation of astrocyte polarization and explore the related mechanism.Methods:Eighty-four male SD rats were randomly assigned to the following groups:A sham operation group,a model group,Taohong Siwutang treatment groups(low dose,medium dose,and high dose),ligustrazine phosphate tablet(LPT)group,and AG490 group.All groups,except for the sham operation group,underwent middle cerebral artery occlusion/reperfusion(MCAO/R)modeling and were treated for seven days.The neurological impairment was evaluated using the Longa score.The volume of cerebral infarction was assessed through 2,3,5-triphenyltetrazolium chloride(TTC)staining.Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)and Western blot analyses were performed to analyze the mRNA and protein expression levels of cortical complement 3(C3),S100 calcium-binding protein A10(S100A10),Janus kinase 2(JAK2),and signal transducer and activator of transcription 3(STAT3).Additionally,protein expression levels of vascular endothelial growth factor-A(VEGF-A)were assessed,and the mRNA expression levels of inflammatory factors,including interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α),were evaluated.Glial fibrillary acidic protein(GFAP)and C3,S100A10 and Co-localization was detected via immunofluorescence double staining.Lastly,VEGF expression levels were measured using enzyme-linked immunosorbent assay(ELISA).Results:Compared with the sham operation group,the model group showed a significant increase in cerebral infarction volume and neurological impairment(P<0.01).C3 protein levels were elevated,while S100A10 levels were decreased.Pathway-related markers were significantly upregulated(P<0.05,P<0.01),and VEGF-A protein levels were significantly reduced(P<0.01).The mRNA expression of inflammatory factors was significantly upregulated(P<0.01).Co-localization analysis showed significantly

关 键 词：桃红四物汤 脑缺血再灌注 炎症因子 星形胶质细胞 非受体型酪氨酸蛋白激酶2(JAK2)/信号转导及转录激活蛋白3(STAT3) 血管内皮生长因子 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R259