肺肠合治法对支气管哮喘小鼠血管活性肠肽和p38 MAPK信号通路的影响  

Effect of Combined Therapy of Lung and Intestine on VIP and p38 MAPK Signaling Pathways in Bronchial Asthma Mice

作　　者：惠毅[1] 张新悦 郑旭锐[1] 闫曙光[1] 史捷 李京涛 HUI Yi;ZHANG Xin-yue;ZHENG Xu-rui;YAN Shu-guang;SHI Jie;LI Jing-tao(College of Basic Medicine,Shaaxi University of Chinese Medicine,Xianyang 712046,China;Affiliated Hospital,Shaaxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学基础医学院,陕西咸阳712046 [2]陕西中医药大学附属医院,陕西咸阳712000

出　　处：《中国实验方剂学杂志》2022年第8期108-115,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81703974);陕西中医药大学学科创新团队建设项目(2019-YL05)。

摘　　要：目的:观察中医治疗肺系疾病的常用治法肺肠合治法对支气管哮喘小鼠的治疗作用,进一步检测与哮喘发病机制密切相关的血管活性肠肽(VIP)和p38丝裂原活化蛋白激酶(p38 MAPK)信号通路相关蛋白的变化,以阐明肺肠合治法治疗支气管哮喘的作用机制。方法:将60只昆明种小鼠随机选12只作为正常组。其余小鼠通过卵清蛋白致敏复制小鼠支气管哮喘模型。分为正常组、模型组、地塞米松组(0.5 mg·kg^(-1)·d^(-1))、中药治肺组(2.73 g·kg^(-1)·d^(-1))、肺肠合治组(6.825 g·kg^(-1)·d^(-1)),灌胃30 d后取血清及肺组织样本。酶联免疫吸附测定法(ELISA)检测各组小鼠血清中VIP、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量,实时荧光定量聚合酶链式反应(Real-time PCR)法检测各组小鼠肺组织中TNF-α、IL-6、p38 MAPK m RNA的表达,蛋白免疫印迹法(Western blot)检测各组小鼠肺组织中TNF-α、IL-6、p38 MAPK,磷酸化p38 MAPK(p-p38 MAPK)蛋白的表达。结果:与正常组比较,模型组小鼠血清VIP含量明显降低(P<0.05),TNF-α、IL-6含量明显升高(P<0.05),肺组织TNF-α、IL-6、p38 MAPK m RNA和TNF-α、IL-6、p-p38 MAPK/p38 MAPK蛋白相对表达量明显升高(P<0.05);与模型组比较,各治疗组小鼠血清VIP含量明显升高(P<0.05),TNF-α、IL-6含量明显升高(P<0.05),肺组织TNF-α、IL-6、p38 MAPK m RNA和TNF-α、IL-6、p-p38 MAPK/p38 MAPK蛋白相对表达量明显降低(P<0.05);与肺肠合治组比较,地塞米松组小鼠血清TNF-α、IL-6明显增高(P<0.05),肺组织TNF-α、IL-6 m RNA和TNF-α、IL-6蛋白相对表达量明显降低(P<0.05),肺组织p38MAPK、VIP m RNA和p-p38 MAPK/p38 MAPK蛋白相对表达量明显升高(P<0.05),中药治肺组小鼠血清VIP、TNF-α、IL-6明显降低(P<0.05),肺组织TNF-α、IL-6、p38 MAPK m RNA和TNF-α、IL-6、p-p38 MAPK/p38 MAPK蛋白相对表达量均明显升高(P<0.05),肺组织VIP m RNA表达水平明显降低(P<0.05)。结论:肺肠合治法可Objective:To observe the therapeutic effects of the combined therapy of lung and intestine,a common treatment for pulmonary diseases in traditional Chinese medicine(TCM),on bronchial asthma mice,and further detect the changes of vasoactive intestinal peptide(VIP)and p38 mitogen-activated protein kinase(p38 MAPK)signaling pathway-related proteins which are closely related to the pathogenesis of asthma,in order to elucidate the mechanism of the combined therapy of lung and intestine in the treatment of bronchial asthma.Method:A total of 60 Kunming mice were randomly divided into normal group,model group,dexamethasone group(0.5 mg·kg^(-1)·d^(-1)),TCM group(2.73 g·kg^(-1)·d^(-1)),and lung-intestine treatment group(6.825 g·kg^(-1)·d^(-1)),12 mice in each group.All mice except the normal group were sensitized by ovalbumin to induce bronchial asthma.After 30 days of intragastric administration,serum and lung tissue samples were obtained.The content of VIP,interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the serum of mice in each group was detected by enzyme-linked immunosorbent assay(ELISA).The m RNA levels of TNF-α,IL-6,and p38 MAPK in lung tissues of mice were detected by real-time quantitative polymerase chain reaction(Real-time PCR),and the protein levels of TNF-α,IL-6,p38 MAPK,and phosphorylated p38 MAPK(p-p38MAPK)in lung tissues of mice were assayed by Western blot(WB).Result:Compared with the normal group,the model group showed decreased content of serum VIP(P<0.05),increased content of TNF-αand IL-6(P<0.05),up-regulated m RNA levels of TNF-α,IL-6,and p38 MAPK,and elevated protein levels of TNF-α,IL-6,and p-p38 MAPK/p38 MAPK in lung tissues(P<0.05).Compared with the model group,the treatment groups exhibited increased content of serum VIP,TNF-α,and IL-6(P<0.05),down-regulated m RNA levels of TNF-α,IL-6,and p38 MAPK,and lower protein levels of TNF-α,IL-6,and p-p38 MAPK/p38 MAPK in lung tissues(P<0.05).As compared with the lung-intestine treatment group,the serum TNF-αand IL-6 levels in the

关 键 词：肺肠合治法 血管活性肠肽 p38丝裂原活化蛋白激酶(p38 MAPK)信号通路 支气管哮喘 三拗汤 小承气汤 

分 类 号：R2-0[医药卫生—中医学] R22