基于Toll样受体4/髓样分化因子88/核转录因子κB信号通路探讨针刺“蝶腭穴”改善变应性鼻炎大鼠免疫炎性反应的机制  

Acupuncture at “Die E acupoint” alleviates inflammatory reaction via inhibiting TLR4/MyD88/NF-κB signaling in rats with allergic rhinitis

作　　者：田美慧 张亚男 孙维芳 刘欢 汤勇 TIAN Mei-hui;ZHANG Ya-nan;SUN Wei-fang;LIU Huan;TANG Yong(College of Traditional Chinese Medicine,Changchun University of Chinese Medicine,Changchun 130117,China;College of Clinical Medicine,Changchun University of Chinese Medicine,Changchun 130117;The Affiliated Hospital of Changchun University of Chinese Medicine,Changchun 130021)

机构地区：[1]长春中医药大学中医学院,长春130117 [2]长春中医药大学临床医学院,长春130117 [3]长春中医药大学附属医院,长春130021

出　　处：《针刺研究》2024年第5期456-462,共7页Acupuncture Research

基　　金：吉林省自然科学基金项目(No.20210101194JC)。

摘　　要：目的:探讨针刺“蝶腭穴”对变应性鼻炎(AR)大鼠Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核转录因子κB(NF-κB)、转录因子T-bet(T-bet)、GATA结合蛋白-3(GATA-3)的蛋白表达水平及相关炎性细胞因子的影响。方法:SD大鼠随机分为空白组、模型组、针刺组、假针刺组,每组6只。采用卵清蛋白诱导建立AR大鼠模型。针刺组大鼠行双侧“蝶腭穴”针刺,假针刺组仅行假针刺,均每日1次,共6次。记录各组大鼠行为学评分;HE染色法观察大鼠鼻黏膜形态变化;ELISA法检测大鼠血清免疫球蛋白E(IgE)、卵清蛋白特异性IgE(OVA-sIgE)、干扰素(IFN)-γ、白细胞介素(IL)-4、IL-10、IL-17的含量;Western blot法检测大鼠鼻黏膜中TLR4、MyD88、NF-κB p65、T-bet、GATA-3蛋白表达水平。结果:与正常组比较,模型组行为学评分升高(P<0.05),血清IgE、OVA-sIgE、IL-4、IL-17的含量及鼻黏膜中GATA-3、TLR4、MyD88、NF-κB p65蛋白表达水平升高(P<0.05),血清IFN-γ、IL-10含量及鼻黏膜T-bet蛋白表达水平降低(P<0.05)。与模型组比较,针刺组行为学评分降低(P<0.05),血清IgE、OVA-sIgE、IL-4、IL-17的含量及鼻黏膜中GATA-3、TLR4、MyD88、NF-κB p65蛋白表达水平降低(P<0.05),血清IFN-γ、IL-10的含量及鼻黏膜T-bet蛋白表达水平升高(P<0.05);假针刺组以上指标差异均无统计学意义。模型组和假针刺组可见鼻黏膜炎性浸润,针刺组鼻黏膜炎性反应明显减轻。结论:针刺“蝶腭穴”可减轻OVA诱导的AR大鼠的症状,改善鼻黏膜炎性反应,其机制可能与抑制TLR4/MyD88/NF-κB信号通路,有效调控Th1/Th2、Treg/Th17细胞因子及T-bet/GATA-3的平衡有关。Objective To observe effects of acupuncture at “Die E acupoint” on the protein expression levels of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),nuclear transcription factor κB(NF-κB),transcription factor T-bet(T-bet),and GATA-binding protein-3(GATA-3) in the nasal mucosa and the serum contents of related inflammatory cytokines in rats with allergic rhinitis,so as to explore the mechanism of acupuncture in treating allergic rhinitis.Methods Twenty-four healthy SD rats were randomly divided into blank,model,acupuncture,and sham acupuncture groups,with 6 rats in each group.The rat model of allergic rhinitis was established by using ovalbumin induction.The rats in the acupuncture group received bilateral acupuncture at the “Die E acupoint” with a depth of 15—20 mm,while the rats in the sham acupuncture group received only sham acupuncture(light and shallow acupunture of the skin at the “Die E acupoint”).Both interventions were performed once daily for a total of 6 days.Behavioral scores of rats in each group were recorded.Pathological changes of nasal mucosa were observed by H.E.staining.Serum contents of IgE,ovalbumin-specific IgE(OVA-sIgE),interferon(IFN)-γ,interleukin(IL)-4,IL-10 and IL-17 were measured by ELISA and the protein expression levels of T-bet,GATA-3,TLR4,MyD88 and NF-κB p65 in the nasal mucosa were detected by Western blot.Results After modeling,compared with the blank group,rats in the model group showed increased behavioral scores,serum IgE,OVA-sIgE,IL-4,and IL-17 contents,and nasal mucosal GATA-3,TLR4,MyD88,and NF-κB p65 protein expression levels(P<0.05),whereas the contents of serum IFN-γ,IL-10 and the protein expression level of T-bet in the nasal mucosa were decreased(P<0.05).Comparison between the EA and model groups showed that acupuncture intervention can decrease the behavioral scores of rats with allergic rhinitis,the contents of serum IgE,OVA-sIgE,IL-4,IL-17,and the protein expression levels of GATA-3,TLR4,MyD88,and NF-κB p65 in the nasal mucosa(

关 键 词：蝶腭穴 变应性鼻炎 免疫炎性反应 Toll样受体4/髓样分化因子88/核转录因子κB信号通路 

分 类 号：R24[医药卫生—中医临床基础]