基于ERK1/2信号通路的加味过敏煎对过敏性哮喘小鼠影响的机制研究  

Exploring the mechanism of Jiawei Guomin Decoction on allergic asthma mice based on the ERK1/2 signaling pathway

作　　者：黄宝萱 林晓彤 张颖琳 覃骊兰[1] HUANG Bao-xuann;LIN Xiao-tong;ZHANG Ying-lin;QIN Li-lan(Guangxi University of Chinese Medicine,Nanning 530200,China;Affiliated Hospital of Guilin Medical University,Guilin 541001,China;Wuzhou Hospital of Traditional Chinese Medicine,Wuzhou 543002,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]桂林医学院附属医院,广西桂林541001 [3]梧州市中医医院,广西梧州543002

出　　处：《时珍国医国药》2025年第2期246-252,共7页Lishizhen Medicine and Materia Medica Research

基　　金：广西自然科学基金面上项目(2023JJA141223);南宁市科学研究与技术开发重点研发计划(20233062);广西中医药大学“桂派杏林拔尖人才”项目(2022C006);广西中医药大学校级重点课题(2021ZD002)。

摘　　要：目的通过卵清蛋白(OVA)诱导建立过敏性哮喘小鼠模型,探讨加味过敏煎对过敏性哮喘小鼠的影响及可能的作用机制。方法通过卵清蛋白(OVA)诱导建立过敏性哮喘小鼠模型,分为模型组、地塞米松阳性组、加味过敏煎高、中、低剂量组。另设不造模的空白组。实验第1、8、15天,除空白组外各组小鼠腹腔注射OVA致敏液,空白组用等量PBS替代;实验第16天开始灌胃给药,连续14d;实验第25~29天,每天灌胃1 h后,除空白组外,各组小鼠进行OVA雾化激发,空白组用PBS替代;实验第30天,进行指标的采集。苏木精-伊红(HE)染色观察小鼠肺组织病理变化;酶联免疫吸附测定(ELISA)测定小鼠血清中卵清蛋白特异性免疫球蛋白E(OVA-sIgE)、白介素-5(IL-5)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)和嗜酸性粒细胞趋化因子(Eotaxin)的含量;RT-qPCR检测小鼠肺组织中细胞外调节蛋白激酶(ERK)1、ERK2、CC趋化因子受体3(CCR3)、Eotaxin mRNA表达水平;采用Western Blot检测小鼠肺组织中ERK1/2、CCR3、Eotaxin的蛋白表达水平。结果与模型组比较,加味过敏煎各剂量小鼠肺组织炎性细胞浸润、支气管上皮细胞脱落等现象有所减轻;高、中剂量组小鼠血清中OVA-sIgE、IL-5含量降低(P<0.01,P<0.05),加味过敏煎各剂量组GM-CSF、Eotaxin含量降低(P<0.01,P<0.05);中、低剂量组小鼠肺组织ERK1基因表达量下降(P<0.01),加味过敏煎各剂量组小鼠肺组织ERK2、CCR3基因表达量下降(P<0.01);高剂量组小鼠肺组织ERK1/2蛋白表达下降(P<0.05),加味过敏煎各剂量组CCR3、Eoatxin蛋白表达下降(P<0.01,P<0.05)。结论加味过敏煎能够改善过敏性哮喘小鼠气道炎症反应,发挥治疗过敏性哮喘的作用,其机制可能与抑制嗜酸性粒细胞的趋化、活化,下调ERK1/2、CCR3、Eotaxin基因和蛋白的表达有关。Objective An ovalbumin(OVA)-induced allergic asthma mice model was established to investigate the effect of Jiawei Guomin Decoction(JGD)on allergic asthma mice and its possible mechanism.Methods The mice model of allergic asthma was induced by OVA and divided into the model group,the dexamethasone-positive group,and JGD high-dose(JGD-H),medium-dose(JGD-M),and low-dose(JGD-L)groups.Additionally,Set up a blank group without mold making.On days 1,8 and 15,mice in each group except the blank group were injected intraperitoneally with OVA allergic solution,and the blank group was replaced by an equal amount of PBS;from day 16 onwards,the mice were gavaged for 14 consecutive days;on days 25~29,after 1 h of gavage each day,mice in each group except the blank group were inspired by OVA nebulisation,and the blank group was replaced by PBS;on day 30,the indexes were collected.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in the lung tissue of mice;enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of OVA-sIgE,interleukin-5(IL-5),granulocyte-macrophage colony-stimulating factor(GM-CSF),and eosinophil chemotactic factor(eotaxin)in the lung tissue of mice.RT-qPCR was used to detect the levels of extracellular regulated protein kinase(ERK)1,ERK2,CC chemokine receptor 3(CCR3),and Eotaxin mRNA;Western blot(WB)was used to detect the protein expression levels of ERK1/2,CCR3,and Eotaxin in the lung tissues of mice.Results Compared with the model group,inflammatory cell infiltration and bronchial epithelial cell detachment in lung tissues of mice in the JGD-H,JGD-M and JGD-L groups were attenuated;serum levels of OVA-sIgE and IL-5 were decreased in mice in the JGD-H and JGD-M groups(P<0.01,P<0.05),and levels of GM-CSF and Eotaxin in mice in each JGD group were decreased(P<0.01,P<0.05),ERK1 gene expression in the lung tissue of mice in the JGD-M and JGD-L groups were decreased(P<0.01),ERK2 and CCR3 gene expression in the lung tissue of mice in each JGD group were decreased(P<0.01);ER

关 键 词：ERK1/2信号通路 加味过敏煎 过敏性哮喘 机制 

分 类 号：R285.5[医药卫生—中药学]