JNK/AP-1信号通路在百草枯诱导小胶质细胞NLRP3炎症小体活化中的作用  

Role of JNK/AP-1 signaling pathway in paraquat-induced activation of NLRP3 inflammasome in microglia

作　　者：兰孟菊 王楷栋 黄敏[1,2] LAN Mengju;WANG Kaidong;HUANG Min(Department of Occupational Health and Environmental Hygiene,School of Public Health,Yinchuan,Ningxia 750004,China;Ningxia Key Laboratory of Environmental Factors and Chronic Disease Control,Ningxia 750004,China)

机构地区：[1]宁夏医科大学公共卫生学院职业卫生与环境卫生系,宁夏银川750004 [2]宁夏环境因素与慢性病控制重点实验室,宁夏银川750004

出　　处：《环境与职业医学》2023年第6期705-710,共6页Journal of Environmental and Occupational Medicine

基　　金：中国科学院“西部之光”人才培养计划(XAB2020YW14);宁夏医科大学科学研究基金资助项目(XZ2021002)。

摘　　要：[背景]环境化学物百草枯(PQ)是一种能够引起散发性帕金森病(PD)的环境因素之一,其中小胶质细胞介导的神经炎症反应在PD发生发展中发挥重要作用。前期研究发现低剂量PQ可将BV-2小胶质细胞激活为M1表型并发挥促炎作用,但这种激活的小胶质细胞介导的神经炎症反应机制尚不清楚。[目的]探讨c-Jun氨基酸末端激酶(JNK)/激活蛋白-1(AP-1)信号通路在PQ诱导小胶质细胞NOD样受体热蛋白结构域相关蛋白(NLRP3)炎症小体活化中的作用。[方法]建立体外BV-2小胶质细胞模型,采用0、0.03、0.06、0.12μmol·L^(-1)PQ处理细胞24 h后提取细胞全蛋白,通过免疫印迹法测定JNK、AP-1组成蛋白(c-Jun、c-Fos)、NLRP3、含胱天蛋白酶募集结构域凋亡相关斑点蛋白(ASC)、胱天蛋白酶-1前体(pro caspase-1)、白介素18(IL-18)、白介素-1β(IL-1β)的相对表达水平,以观察PQ暴露对JNK/AP-1信号通路和NLRP 3炎症小体的影响。通过20μmol·L^(-1)JNK抑制剂SP600125干预后,再次检测上述蛋白以探究JNK/AP-1信号通路对NLRP3炎症小体活化的驱动作用。[结果]PQ暴露后,0.06μmol·L^(-1)PQ组和0.12μmol·L^(-1)PQ组JNK、c-Jun、c-Fos、NLRP3、ASC、pro caspase-1的表达水平高于0μmol·L^(-1)PQ组(P<0.05),炎症因子(IL-18、IL-1β)的相对表达水呈逐渐增加(P<0.05)。经JNK抑制剂SP600125干预后,对照组、20μmol·L^(-1)SP600125组、20μmol·L^(-1)SP600125+0.06μmol·L^(-1)PQ组JNK/AP-1信号通路关键蛋白、NLRP3炎症小体关键蛋白及IL-18、IL-1β的相对表达水平低于0.06μmol·L^(-1)PQ组(P<0.05)。[结论]PQ暴露能够激活JNK/AP-1信号通路驱动BV-2小胶质细胞NLRP3炎症小体激活介导神经炎症反应。[Background]Paraquat(PQ)is one of the environmental factors that can cause sporadic Parkinson's disease(PD).Microglia-mediated neuroinflammation plays an important role in the occurrence and development of PD.Our previous studies have found that low doses of PQ can activate BV-2 microglia to the M1 phenotype and exert pro-inflammatory effects,but the associated mechanism is not clear yet.[Objective]To explore the role of c-Jun N-terminal kinase(JNK)/activator protein 1(AP-1)signaling pathway in PQ-induced activation of the NOD-like receptor thermal protein domain associated protoin 3(NLRP3)inflammasome in microglia.[Methods]An in vitro microglia model was established.The cells were treated with 0,0.03,0.06,and 0.12μmol·L^(-1) PQ for 24 h,the whole cell protein was extracted.The relative expression levels of JNK,AP-1 constituent proteins(c-Jun,c-Fos),NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),caspasse-1 precursor(pro caspase-1),interleukin-18(IL-18),and interleukin-1β(IL-1β)were evaluated by Western blotting,to observe the effects of PQ exposure on JNK/AP-1 signaling pathway and NLRP3 inflammasome.After the treatment of 20μmol·L^(-1) JNK inhibitor SP600125,the above proteins were detected again,to explore the driving effect of JNK/AP-1 signaling pathway on NLRP3 inflammasome activation.[Results]After PQ exposure,the relative expression levels of key proteins of JNK,c-Jun,and c-Fos,NLRP3,ASC,and pro caspase-1 in the 0.06μmol·L^(-1) PQ group and the 0.12μmol·L^(-1) PQ group were higher than those in the 0μmol·L^(-1) PQ group(P<0.05),and the relative expression levels of IL-18 and IL-1βincreased with higher exposure(P<0.05).After the treatment of JNK inhibitor SP600125,the relative expression levels of key proteins of JNK/AP-1 signaling pathway(JNK,c-Jun,and c-Fos),NLRP3 inflammasome(NLRP3,ASC,and Pro caspase-1),and inflammatory factors(IL-18 and IL-1β)in the control group,the 20μmol·L^(-1) SP600125 group,and the 20μmol·L^(-1) SP600125+0.06μmol·L^(-1) PQ group were lower

关 键 词：百草枯 帕金森病 JNK/AP-1 NLRP3炎症小体 BV-2细胞 小胶质细胞 

分 类 号：R114[医药卫生—卫生毒理学]