基于IL-27/STAT1通路探讨针刺肺俞穴纠正哮喘模型大鼠Th1/Th2免疫平衡的机制  

Mechanism of acupuncture at"Feishu"(BL13)to correct Th1/Th2 immune balance in asthma model rats based on IL-27/STAT1 pathway

作　　者：雷俊[1] 罗玲艳 孙艺 乔松[1] LEI Jun;LUO Lingyan;SUN Yi;QIAO Song(Department of Pulmonary Diseases,Wuhan Hospital of Traditional Chinese Medicine,Wuhan,Hubei 430000,China;Medical Department of Yangtze University,Jinzhou,Hubei 433200,China;Oncology Department,ShanghaiGongli Hospital of Pudong District,Shanghai 200135)

机构地区：[1]武汉市中医医院肺系病科,湖北武汉430000 [2]长江大学医学部,湖北荆州433200 [3]上海市浦东新区公利医院肿瘤科,上海200135

出　　处：《湖南中医药大学学报》2022年第6期986-993,共8页Journal of Hunan University of Chinese Medicine

基　　金：武汉市卫生健康委员会医学科研项目(WZ21C57)。

摘　　要：目的从白细胞介素(interleukin,IL)-27/信号转导和转录激活因子1(signal transducer and activator of transcription1,STAT1)探究针刺肺俞穴改善哮喘大鼠辅助性T细胞(helper T cell,Th)1/Th2免疫平衡的可能机制。方法取140只SD雄性大鼠,随机分为正常组、模型组、IL-27预防组、IL-27治疗组、针刺肺俞组、STAT1抑制剂组、针刺肺俞加STAT1抑制剂组,每组20只。除正常组外,其余各组均进行卵白蛋白反复致敏建立哮喘模型。肺功能仪检测气道高反应性;ELISA法检测血清及肺泡灌洗液中IL-4、干扰素γ(interferonγ,IFN-γ)、免疫球蛋白E(immunoglobulin,Ig E)水平;流式细胞仪检测脾脏Th1/Th2比例;HE染色观察肺组织病理变化;免疫组化法检测IL-4、IFN-γ、磷酸化STAT1(p-STAT1)阳性表达;Western blot法检测IL-27、STAT1、IL-4、IFN-γ、T-bet及GATA3蛋白表达。结果与正常组相比,模型组大鼠气道高反应性升高,脾脏Th1/Th2比例降低,血清及肺泡灌洗液中IL-4水平升高而IFN-γ水平降低,肺泡灌洗液中嗜酸性粒细胞比例升高,肺组织支气管黏膜水肿及炎性细胞浸润等病理损伤加重,促Th1/Th2比例平衡相关通路IL-27/STAT1处于抑制状态(P<0.05)。与模型组相比,IL-27预防组、IL-27治疗组及针刺肺俞组均可缓解大鼠肺组织损伤,促进IL-27/STAT1活化,纠正Th1/Th2平衡,改善哮喘大鼠气道高反应性(P<0.05);其中,IL-27预防组对哮喘大鼠的上述改善作用均优于针刺肺俞组(P<0.05)。此外,STAT1抑制剂可加重哮喘大鼠肺组织损伤,促进气道高反应及Th1/Th2免疫平衡紊乱,并削弱针刺肺俞穴发挥的促IL-27/STAT1活化、纠正Th1/Th2免疫平衡等作用(P<0.05)。结论针刺肺俞穴治疗哮喘大鼠的作用机制,可能与促进IL-27/STAT1通路活化,纠正Th1/Th2免疫平衡有关。Objective To explore the potential mechanism of acupuncture at"Feishu"(BL13)to improve the immune balance of helper T cell(Th)1/Th2 in asthma rats based on interleukin(IL)-27/signal transducer and activator of transcription 1(STAT1).Methods A total of 140 male SD rats were randomly divided into normal group,model group,IL-27 prevention group,IL-27treatment group,acupuncture Feishu group,STAT1 inhibitor group and acupuncture Feishu plus STAT1 inhibitor group,with 20rats in each group.Except the normal group,the other groups were repeatedly sensitized with ovalbumin to establish asthma model.Airway hyperresponsiveness was measured by pulmonary function instrument;the levels of interleukin(IL)-4,interferonγ(IFN-γ)and immunoglobulin E(IGE)in serum and alveolar lavage fluid were detected by ELISA;the ratio of Th1/Th2 in spleen was detected by flow cytometry;HE staining was used to observe the pathological changes of lung tissue;positive expression of IL-4,IFN-γand phosphorylated STAT1(p-STAT1)were detected by immunohistochemistry;protein expression levels of IL-27,STAT1,IL-4,IFN-γ,T-bet and GATA3 were detected by Western blot.Results Compared with the normal group,the airway hyperresponsiveness of model group increased,the ratio of spleen Th1/Th2 decreased,the levels of IL-4 in serum and alveolar lavage fluid increased,and the level of IFN-γdecreased,the proportion of eosinophils in alveolar lavage fluid increased,the pathological damage such as bronchial mucosal edema and inflammatory cell infiltration in lung tissue aggravated,and the related pathway of promoting Th1/Th2ratio balance IL-27/STAT1 was inhibited(P<0.05).Compared with the model group,IL-27 prevention group,IL-27 treatment group and acupuncture Feishu group can alleviate lung tissue injury,promote IL-27/STAT1 activation,correct Th1/Th2 balance and improve airway hyperresponsiveness in asthma rats(P<0.05);among them,the above improvement effects of IL-27 prevention group on asthma rats were better than that of acupuncture Feishu group(P<0.05).In addi

关 键 词：哮喘 白细胞介素-27 信号转导和转录激活因子1 免疫平衡 针刺 肺俞穴 

分 类 号：R256.12[医药卫生—中医内科学]