雷公藤甲素通过促进小胶质细胞M2极化减轻脑缺血再灌注大鼠神经元损伤  

Triptolide reduces neuronal damage in cerebral ischemia-reperfusion rats by promoting microglial M2 polarization

作　　者：穆秉桃[1] 郭敏芳[1] 于婧文[1] 张慧宇 MU Bingtao;GUO Minfang;YU Jingwen;ZHANG Huiyu(School of Medicine,Shanxi Datong University,Datong 037009,China;School of Traditional Chinese Medicine and Health Service,Shanxi Datong University,Datong 037009,China)

机构地区：[1]山西大同大学医学院,山西大同037009 [2]山西大同大学中医药健康服务学院,山西大同037009

出　　处：《细胞与分子免疫学杂志》2024年第3期222-228,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：山西省基础研究计划项目(20210302123478,20210302123476,20210302123337)。

摘　　要：目的研究雷公藤甲素(TP)对大鼠脑缺血再灌注(I/R)后小胶质细胞M1/M2极化的影响及分子机制。方法采用大脑中动脉栓塞法(MCAO)制备大脑中动脉栓塞再灌注模型,给予(0.1、0.2)mg/kg TP处理,假手术组大鼠作为对照。Longa评分法进行大鼠神经功能缺损进行评分,HE染色观察缺血侧脑组织神经元形态,神经元特异性核蛋白(NeuN)免疫荧光染色观察神经元数量;Western blot法检测缺血侧脑组织钙离子结合接头蛋白分子1(Iba1)、诱导型一氧化氮合酶(iNOS)、精氨酸酶1(Arg1)、Toll样受体4(TLR4)、核因子κB(NF-κB)、NeuN和胱天蛋白酶3(caspase-3)的表达;ELISA检测白细胞介素1β(IL-1β)和IL-10的水平;免疫荧光双标记法检测小胶质细胞内Arg1和TLR4的表达。结果与模型组比较,TP处理组神经学评分明显降低,神经元损伤明显改善;IL-1β水平降低,IL-10水平增加;iNOS、TLR4、NF-κB、和caspase-3表达降低,Arg1和NeuN表达增加。结论TP处理逆转大鼠脑I/R损伤,可能与促进小胶质细胞M2极化、减少炎症因子释放与抑制凋亡有关。Objective To investigate the effects of triptolide(TP)on microglial M1/M2 polarization after cerebral ischemia-reperfusion(I/R)injury in rats and the underlying molecular mechanism.Methods A rat model of middle cerebral artery occlusion(MCAO)was established.TP was administered to rats at doses of 0.1 and 0.2 mg/kg,with a sham surgery group as the control group.Longa scoring was performed to grade neurological deficits in rats;HE staining was used to observe the morphology of neurons in ischemic brain tissues;neuron-specific nuclear protein(NeuN)immunofluorescence staining was used to measure the number of neurons;and Western blot analysis was used to measure the expression levels of ionised calcium-binding adaptor molecule-1(Iba1),inducible nitric oxide synthase(iNOS),arginase 1(Arg1),Toll-like receptor 4(TLR4),nuclear factorκB(NF-κB),NeuN and caspase-3 in ischemic-brain tissues.The protein levels of interleukin 1β(IL-1β)and IL-10 were measured by ELISA.Immunofluorescence double labelling was performed to detect the expression of Arg1 and TLR4 in microglia.Results Compared with the model group,the neurological score of the TP treatment group was significantly reduced and the neuronal damage was significantly alleviated.IL-1βlevels decreased while IL-10 levels increased.The expression levels of iNOS,TLR4,NF-κB and caspase-3 decreased,while the expression levels of Arg1 and NeuN increased.Conclusion TP treatment ameliorates cerebral I/R injury in rats,which may be attributed to the promotion of microglial M2 polarization,thereby reducing the release of inflammatory factors and inhibiting apoptosis.

关 键 词：雷公藤甲素(TP) 缺血再灌注(I/R) 小胶质细胞 极化 炎症 凋亡 

分 类 号：R285.5[医药卫生—中药学] R743.31[医药卫生—中医学] R743R364.5R392