基于cAMP/PKA/CREB信号通路探讨麻黄生物碱组分对过敏性哮喘大鼠的抗炎机制  

Exploring the Anti-inflammatory Mechanism of Ephedra Herba Alkaloid Components in Allergic Asthmatic Rats Based on cAMP/PKA/CREB Signaling Pathway

作　　者：郑亚娟 袁培培[1,2] 傅阳 高丽媛[1] 魏亚新 李潘营 阮元 陈祎 冯卫生 郑晓珂[1,2,3] ZHENG Yajuan;YUAN Peipei;FU Yang;GAO Liyuan;WEI Yaxin;LI Panying;RUAN Yuan;CHEN Yi;FENG Weisheng;ZHENG Xiaoke(Henan University of Chinese Medicine,Zhengzhou 450046 Henan,China;The Engineering and Technology Center for Chinese Medicine Development,Zhengzhou 450046 Henan,China;Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan&Education Ministry of P.R.China,Zhengzhou 450046 Henan,China)

机构地区：[1]河南中医药大学,河南郑州450046 [2]河南省中药开发工程技术研究中心,河南郑州450046 [3]呼吸疾病中医药防治省部共建协同创新中心,河南郑州450046

出　　处：《中药新药与临床药理》2022年第11期1453-1459,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家重点研发计划-中医药现代化研究项目项目(2019YFC1708802);河南省高层次人才特殊支持计划“中原千人计划”-中原领军人才项目(ZYQR2018100)。

摘　　要：目的 探究麻黄生物碱组分对过敏性哮喘大鼠肺部炎症的作用及机制。方法 将32只雄性SD大鼠随机分为正常组、模型组、麻黄生物碱组(37.79 mg·kg^(-1))、地塞米松组(0.075 mg·kg^(-1))。采用卵清蛋白(OVA)和氢氧化铝凝胶佐剂致敏结合OVA雾化激发的方法建立过敏性哮喘大鼠模型。第1天和第8天,大鼠腹腔注射致敏液(Ⅴ级OVA 10 mg+氢氧化铝10 mg)致敏;第15天至第21天灌胃给药30 min后,用2%Ⅱ级OVA雾化激发40 min。检测大鼠喘息次数和喘息潜伏时间;采用HE染色法观察大鼠肺组织病理变化;ELISA法检测肺泡灌洗液中的白细胞介素4(IL-4)、免疫球蛋白E(IgE)、干扰素γ(IFN-γ)水平,以及肺组织匀浆和血清中的环磷酸腺苷(cAMP)水平;Western Blot法检测肺组织中蛋白激酶A(PKA)、cAMP反应元件结合蛋白(CREB)、IL-6、磷酸化信号传导子及转录激活子3(p-STAT3)蛋白表达水平;RT-qPCR法检测肺组织中IL-6、STAT3、IL-1β、肿瘤坏死因子α(TNF-α)mRNA表达水平。结果 与正常组比较,模型组大鼠的喘息潜伏时间显著缩短(P<0.01),喘息次数、炎症评分显著升高(P<0.01);肺泡灌洗液中的IL-4、IgE水平显著升高(P<0.01),IFN-γ水平显著降低(P<0.01);肺组织匀浆及血清中的cAMP含量明显降低(P<0.05,P<0.01);肺组织中PKA、CREB蛋白表达明显下调(P<0.05),IL-6、p-STAT3蛋白表达显著上调(P<0.01);肺组织中IL-6、STAT3、IL-1β、TNF-α mRNA表达显著上调(P<0.01)。与模型组比较,给药组大鼠的喘息潜伏时间明显延长(P<0.05,P<0.01),喘息次数、炎症评分显著降低(P<0.01);肺泡灌洗液中的IL-4、IgE水平显著降低(P<0.01),IFN-γ水平显著升高(P<0.01);肺组织匀浆及血清中的cAMP含量明显升高(P<0.05,P<0.01);肺组织中PKA、CREB蛋白表达显著上调(P<0.01),IL-6、p-STAT3蛋白表达显著下调(P<0.01);肺组织中IL-6、STAT3、IL-1β、TNF-α mRNA表达显著下调(P<0.01)。结论 麻黄生物碱组分可能通过cAMP/PKA/CREB�Objective To investigate the effect and mechanism of Ephedra Herba alkaloid components on lung inflammation in allergic asthmatic rats.Methods Thirty-two male SD rats were divided into normal group,mode group,Ephera Herba alkaloid group(37.79 mg·kg^(-1))and dexamethasone group(0.075 mg·kg^(-1)).The allergic asthma rat model was established by using ovalbumin(OVA)and aluminium hydroxide gel adjuvant sensitization combined with OVA nebulization excitation.On days 1 and 8,the rats were sensitized by intraperitoneal injection of sensitizing solution(grade V OVA 10 mg+aluminium hydroxide 10 mg);from day 15 to day 21,the rats were nebulised with 2%grade II OVA for 40 minutes after gavage administration for 30 minutes.The wheezing times and the latency time of wheezing in rats were detected;the histopathological changes of lung in rats were observed by HE staining method.The levels of interleukin 4(IL-4),immunoglobulin E(IgE),interferon γ(IFN-γ)in bronchoalveolar lavage fluid,and cyclic adenosine monophosphate(cAMP) in lung-homogenate and serum were detected by ELISA;protein kinase A(PKA),cAMP response element binding protein(CREB),IL-6,phosphorylated signal transducer and activator of transcription 3(p-STAT3) protein expression level were detected by Western Blot;mRNA expression levels of inflammatory factors IL-6,STAT3,IL-1β and tumour necrosis factor alpha(TNF-α)were detected by RT-qPCR.Results Compared with the normal group,the latent time of wheezing in the model group was significantly decreased(P<0.01),the wheezing times and inflammation scores were significantly increased(P<0.01);the levels of IL-4 and IgE in bronchoalveolar lavage fluid were significantly increased(P<0.01),while the level of IFN-γ was significantly decreased(P<0.01).The contents of cAMP in lung tissue homogenate and serum were significantly decreased(P<0.05,P<0.01).The protein expressions of PKA and CREB in lung tissue were significantly down-regulated(P<0.05),while the protein expressions of IL-6 and p-STAT3 were significantly upregulat

关 键 词：麻黄生物碱组分 过敏性哮喘 炎症 cAMP/PKA/CREB信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]