阿那其根醇提物通过JNK、p-JNK/p38MAPK信号通路调控咳嗽变异性哮喘大鼠脑部神经炎症的作用研究  

Effect of ethanol extract of root of Anacyclus pyrethrum on regulating neuroinflammation in brain of cough variant asthma rats through JNK and p-JNK/p38MAPK signaling pathways

作　　者：陈伟 杨浩[1] 金小越[1] CHEN Wei;YANG Hao;JIN Xiaoyue(Department of Pharmacy,the Sixth Affiliated Hospital of Xinjiang Medical University,Urumqi 830002,China)

机构地区：[1]新疆医科大学第六附属医院药学部,新疆乌鲁木齐830002

出　　处：《药物评价研究》2024年第11期2559-2566,共8页Drug Evaluation Research

基　　金：新疆维吾尔自治区自然科学基金面上项目(2021D01C458)。

摘　　要：目的探讨阿那其根醇提物(EEAP)对咳嗽变异性哮喘(CVA)大鼠神经炎症和c-Jun N-终端激酶(JNK)/p38丝裂原活化蛋白激酶(p38MAPK)信号通路的调控作用。方法体外实验中,将大鼠海马神经元H19-7细胞分为6组,分别为对照组、模型组、醋酸泼尼松(0.025 mg·mL^(−1))阳性药对照组和EEAP高、中、低质量浓度(6.4、3.2、1.6 mg·mL^(−1))组,采用脂多糖(LPS)诱导细胞炎症模型,采用酶联免疫吸附(ELISA)法检测各组细胞上清液中的白细胞介素-6(IL-6)、IL-8、IL-18的水平,Western blotting法检测各组神经元细胞中JNK、p-JNK和p38MAPK的蛋白表达。动物实验中,将SPF级雄性SD大鼠随机分为对照组、模型组、醋酸泼尼松(2.5 mg·kg^(−1))组和EEAP高、中、低剂量(640、320、160 mg·kg^(−1))组,除对照组外,各组大鼠使用1 mg OVA致敏,大鼠sc新鲜配制的1 mg·mL^(−1)的OVA溶液,在各大鼠后足跖、腹股沟、腰部、背部和颈部总共取10个点,每个点注射0.05 mL,同时ip 0.5 mL,共计l mL;造模后第15天开始,大鼠持续雾化吸入1%OVA 15 d,每天1次,每次20 min以激发哮喘。对照组以0.9%氯化钠溶液代替OVA溶液进行sc和雾化吸入。从造模第30天开始,对照组和模型组ig给予等量0.9%氯化钠溶液,各给药组分别ig给予相应剂量药物,每日1次,持续30 d。采用免疫组织化学法检测海马区组织中JNK和p38MAPK的表达,采用Western blotting法检测海马区组织中p-JNK的蛋白表达,采用ELISA法检测各组大鼠海马区组织中IL-6、IL-8、IL-18的水平。结果体外实验显示,与对照组比较,模型组细胞培养上清液中的IL-6、IL-8、IL-18水平明显升高(P<0.05),模型组LPS刺激的神经元细胞中的p-JNK、p38MAPK蛋白表达水平明显升高(P<0.05);与模型组比较,醋酸泼尼松组及EEAP高、中剂量组细胞培养上清液中的IL-6、IL-8、IL-18水平明显降低(P<0.05),细胞中p-JNK、p38MAPK蛋白表达水平明显降低(P<0.05);且EEAP的炎症因子�Objective To investigate the regulatory effects of(ethanol extract of the root of Anacyclus pyrethrum,EEAP)on neuroinflammation and c-Jun N-terminal kinase(JNK)/p38 mitogen-activated protein kinase(p38MAPK)signaling pathway in cough-variant asthma(CVA)rats.Methods In in vitro experiments,rat hippocampal neuronal cells H19-7 were divided into six groups,namely,the control group,the model group,the prednisone acetate(0.025 mg·mL^(−1))positive drug control group,and the EEAP high,medium,and low mass concentration(6.4,3.2,and 1.6 mg·mL^(−1))group,and lipopolysaccharide(LPS)was used to induced cellular inflammation model,and the levels of interleukin-6(IL-6),IL-8,and IL-18 in the cell supernatants of each group were detected by enzyme-linked immunosorbent assay(ELISA),and the protein expression of JNK,p-JNK,and p38MAPK in the neuronal cells of each group was detected by Western blotting.In the animal experiments,SPF-grade male SD rats were randomly divided into control,model,prednisone acetate(2.5 mg·kg−1),and EEAP high,medium,and low dose(640,320,and 160 mg·kg−1)groups,and rats in each group except the control group were sensitized with 1 mg of OVA,and the rats sc freshly prepared 1 mg·mL^(−1) OVA solution was injected at a total of 10 points in the hindfoot plantar,inguinal,lumbar,dorsal,and cervical regions of each rat,with 0.05 mL injected at each point,while 0.5 mL was injected intraperitoneally,for a total of l mL.Starting on the 15th d after modeling,the rats were continuously nebulized and inhaled with 1%OVA for 15 d,once per day,for 20 min each time in order to stimulate asthma.The control group was sc and nebulized inhalation with 0.9%NaCl solution instead of OVA solution.Starting from the 30th d of modeling,the control group and the model group ig were given equal amounts of 0.9%NaCl solution,and each dosing group ig was given the corresponding dose of drug once a day for 30 d.The expression of JNK and p38MAPK in hippocampal tissues was detected by immunohistochemistry,and the protein express

关 键 词：阿那其根醇提物 咳嗽变异性哮喘 神经炎症反应 c-Jun N-终端激酶/p38丝裂原活化蛋白激酶信号通路 白细胞介素 海马神经元 

分 类 号：R285.5[医药卫生—中药学]