基于TLR4-NF-κB通路的槲皮素对病毒性呼吸道感染小鼠免疫功能的影响及抑炎作用研究  

Quercetin Improves Immune Function and Alleviates Inflammation in Mice with Viral Respiratory Tract Infection via TLR4/NF-κB Signaling Pathway

作　　者：许玲芬[1] 李英兰[1] 王佳斌[1] 于湧[1] 顾玉海[1] XU Lingfen;LI Yinglan;WANG Jiabin;YU Yong;GU Yuhai(Department of General Medicine,Qinghai Provincial People's Hospital,Xining 810000)

机构地区：[1]青海省人民医院全科医学科,西宁810000

出　　处：《中药药理与临床》2023年第6期53-57,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：青海省科技计划项目(编号:2019-SF-L4)。

摘　　要：目的:探讨槲皮素对病毒性呼吸道感染小鼠免疫功能的影响及对Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路的调节作用。方法:60只小鼠采用呼吸道合胞病毒(RSV)滴鼻感染法建立呼吸道感染模型,3 d后,按照随机数字表法将小鼠分为正常对照组、模型对照组、槲皮素50、100 mg/kg组和利巴韦林150 mg/kg组,各组灌胃相应药物或生理盐水,1次/d,连续14 d。流式细胞仪检测T辅助17(Th17)细胞和调节性T(Treg)细胞浓度,检测肺指数和脾指数;ELISA法测定高敏C反应蛋白(HS-CRP)、肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)、白细胞介素-17(IL-17)、IL-23和IL-10含量;HE染色观察肺组织病理损伤;蛋白印迹法检测TLR4、NF-κB和髓样分化因子88(MYD 88)蛋白相对表达。结果:正常对照组小鼠肺组织肺泡形态正常,未见炎性细胞浸润;模型对照组小鼠肺泡结构紊乱,肺泡腔增厚,可见大量炎性细胞浸润,并出现组织出血,利巴韦林150 mg/kg组肺组织结构较完整,肺泡结构较明显,无出血,可见少量炎性细胞浸润,槲皮素50、100 mg/kg组肺损伤均不同程度减轻。与正常对照组比较,模型对照组Th17细胞浓度分数、肺指数和脾指数及肺组织HS-CRP、TNF-α、IFN-γ、IL-17、IL-23含量明显升高,TLR4、NF-κB、MYD 88蛋白相对表达明显上调,Treg细胞浓度分数和IL-10含量明显降低(P<0.05);与模型对照组比较,槲皮素50、100 mg/kg组及利巴韦林150 mg/kg组Th17细胞浓度分数、肺指数和脾指数、肺组织HS-CRP、TNF-α、IFN-γ、IL-17、IL-23含量明显降低、TLR4、NF-κB、MYD 88蛋白相对表达明显下调,Treg细胞浓度分数和IL-10含量明显升高(P<0.05)。结论:槲皮素可改善病毒性呼吸道感染小鼠免疫功能,抑制炎症反应,减轻肺损伤,可能是通过TLR4/NF-κB信号通路发挥调节作用。Objective:To investigate the effects of quercetin on the immune function and Toll-like receptor 4(TLR4)/nuclear factor κB(NF-κB) signaling pathway in mice with viral respiratory tract infection.Methods:The respiratory tract infection model of 60 mice was established by nasal dripping of respiratory syncytial virus.After 3 days,mice were assigned into a control group,a model group,two quercetin groups(50 mg/kg and 100 mg/kg),and a ribavirin(150 mg/kg) group according to the random number table method.Corresponding drugs or saline water were administrated intragastrically once a day for 14 consecutive days.The T-assisted 17(Th17) cells and regulatory T(Treg) cells were detected by flow cytometry,and then lung index and spleen index were calculated.The levels of high-sensitivity C-reactive protein(HS-CRP),tumor necrosis factor-alpha(TNF-α),interferon-gamma(IFN-γ),interleukin-17(IL-17),IL-23,and IL-10 were determined by enzyme-linked immunosorbent assay(ELISA).The pathological injury of the lung tissue was observed by hematoxylin-eosin(HE) staining.The relative expression levels of TLR4,NF-κB,and myeloid differentiation factor 88(MyD88) were measured by Western blotting.Results:The mice in the control group showed normal morphology of pulmonary alveoli and no inflammatory cell infiltration.The mice in the model group showed disorganized alveolar structure,thickening alveolar cavity,a large number of inflammatory cell infiltration,and tissue bleeding.The mice in the ribavirin(150 mg/kg) group had intact lung tissue structure,obvious alveolar structure,no bleeding,and a small amount of inflammatory cell infiltration.Quercetin(50 mg/kg and 100 mg/kg) alleviated the lung injury to varying degrees.Compared with the control group,the modeling increased the Th17 cell proportion,lung index,and spleen index,elevated the levels of HS-CRP,TNF-α,IFN-γ,IL-17,and IL-23,up-regulated the expression of TLR4,NF-κB,and MyD88,and reduced the Treg cell proportion and IL-10 level(P<0.05).Compared with the model group,quercetin(50

关 键 词：槲皮素 病毒性呼吸道感染 免疫功能 Toll样受体4 核因子-ΚB 髓样分化因子88 

分 类 号：R285.5[医药卫生—中药学]