基于TLR2/MyD88/NF-κB信号通路探讨不可分型流感嗜血杆菌诱导支气管上皮细胞炎症的机制  

Mechanism of inflammation in bronchial epithelial cells induced by Nontypeable Hemophilus influenzae based on TLR2/MyD88/NF-κB Pathways

作　　者：周欣欣[1] 张晓宇[2] 周辉[1] 王小妹[1] Zhou Xinxin;Zhang Xiaoyu;Zhou Hui;Wang Xiaomei(the First Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410000,China;the Second People's Hospital of Hunan Province,Changsha Hunan 410000,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410000 [2]湖南省第二人民医院手足外科,湖南长沙410000

出　　处：《遵义医科大学学报》2021年第3期314-318,325,共6页Journal of Zunyi Medical University

基　　金：湖南省卫生健康委基金资助项目(NO:20201327);湖南省教育厅基金资助项目(NO:19C1425,20C1427);湖南省自然科学青年基金资助项目(NO:2020JJ5438)。

摘　　要：目的初步探讨不可分型流感嗜血杆菌(NTHi)对支气管上皮细胞炎症反应的影响及其分子机制。方法体外培养人正常支气管上皮细胞(BEAS-2B),采用感染复数MOI=1、5、10的NTHi感染BEAS-2B细胞后,ELISA、qRT-PCR检测细胞上清中IL-1β和IL-6及其mRNA的表达水平,并检测细胞内TLR2、MyD88的活化情况以及IκB的磷酸化水平;随后采用siRNA以及MyD88、NF-κB抑制剂预处理细胞,并检测细胞中IL-1β和IL-6的表达水平。结果MOI=1、5、10的NTHi刺激BEAS-2B细胞后,细胞内IL-1β和IL-6及其mRNA水平均显著增高(P<0.05),且TLR2以及MyD88的表达水平也要显著高于未刺激组(P<0.05)。Western blot结果显示,NTHi处理后细胞内IκB的磷酸化水平较未刺激组明显上升;siRNA沉默TLR2及采用NF-κB抑制剂、MyD88抑制剂后细胞中IL-1β和IL-6分泌水平较对照组显著降低(P<0.05)。结论NTHi能够通过激活TLR2/MyD88以及NF-κB信号通路诱导支气管上皮细胞BEAS-2B分泌促炎因子IL-1β以及IL-6。Objective To investigate the effects of Nontypeable Hemophilus influenzae(NTHi)on the inflammatory response of bronchial epithelial cells and its molecular mechanism.Methods Human normal bronchial epithelial cells(BEAS-2B)were cultured in vitro,and the expression levels of IL-1βand IL-6 in the BEAS-2B cell were detected by ELISA and qPCR after infected with NTHi with plural MOI=1,5,10.And then the activation of TLR2 and the phosphorylation level of IκB were detected.The expression levels of IL-1βand IL-6 were detected before the cells were pretreated with siRNA and MyD88,NF-κB inhibitors.Results After the treatment with NTHi of MOI=1,5,10 in BEAS-2B cells,the levels of IL-1βand IL-6 in the cells were significantly increased,and the expression levels of TLR2 and MyD88 were also significantly higher than that of the unstimulated group.Western blot results showed that the phosphorylation level of IκB was significantly increased after NTHi treatment compared with that of the non-stimulated group.The levels of IL-1 and IL-6 in TLR2 silenced with siRNA,pretreatment with NF-κB inhibitor and MyD88 inhibitor were significantly lower than those in control group.Conclusion NTHi can induce to the secretion of pro-inflammatory cytokines IL-1βand IL-6 in bronchial epithelial cells BEAS-2B via activating TLR2/MyD88/NF-κB signaling pathways.

关 键 词：不可分型流感嗜血杆菌 TLR2 NF-ΚB 炎症因子 

分 类 号：R392[医药卫生—免疫学]