羟氯喹通过Toll样受体4/核因子-κB信号通路对系统性红斑狼疮小鼠肺损伤的影响  

Effect of hydroxychloroquine on lung injury in mice with systemic lupus erythematosus via TLR4/NF-κB signaling pathway

作　　者：付永涛 王莉平[1] 郭金明[1] 张娜[2] 张红霞[2] 许冰 FU Yongtao;WANG Liping;GUO Jinming;ZHANG Na;ZHANG Hongxia;XU Bing(Handan Central Hospital,Handan 056001,China)

机构地区：[1]邯郸市中心医院,河北邯郸056001 [2]石家庄市第四医院,河北石家庄050035 [3]邯郸市邯钢医院,河北邯郸056001

出　　处：《陕西医学杂志》2023年第11期1483-1488,共6页Shaanxi Medical Journal

基　　金：河北省重点研发计划项目(172777231);河北省石家庄市科学技术研究与发展计划项目(201200773)。

摘　　要：目的:探讨羟氯喹(HCQ)对系统性红斑狼疮(SLE)小鼠肺损伤以及Toll样受体4/核因子-κB(TLR4/NF-κB)信号通路的影响。方法:取32只雌性MRL/lpr狼疮小鼠(自发型SLE小鼠模型)随机分为模型组、HCQ(80 mg/kg)组、HCQ(80 mg/kg)+TAK242(TLR4抑制剂,0.3 mg/kg)组和HCQ(80 mg/kg)+LPS(TLR4激动剂,2.5 mg/kg)组,每组8只;另取8只同龄雌性C57BL/6小鼠设为正常对照组。各组分别连续给药治疗5周后,通过肺功能仪检测肺功能指标[气道阻力(Raw)、动态肺顺应性(Cdyn)、第0.1秒用力呼气容积(FEV0.1)],ELISA法检测肺泡灌洗液中炎症因子[干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、IL-1β、IL-6]水平和血清中免疫学指标[抗核抗体(ANA)、抗双链DNA(ds-DNA)]水平,HE染色法和Masson染色法观察肺组织病理学改变和纤维化状况,并进行病理评分和计算胶原容积分数(CVF),RT-qPCR法和Western blot法检测肺组织TLR4、NF-κB p65、IκBα mRNA和蛋白表达。结果:与模型组比较,HCQ组小鼠Raw明显降低且Cdyn、FEV0.1明显升高(均P<0.05);肺泡灌洗液IFN-γ、TNF-α、IL-1β、IL-6水平和血清ANA、ds-DNA水平明显降低(均P<0.05);肺组织病理学改变和纤维化状况明显改善,病理评分和CVF明显降低(均P<0.05);肺组织TLR4、NF-κB p65、IκBα mRNA和蛋白表达量均明显降低(均P<0.05)。TAK242能够明显增强HCQ对SLE小鼠各检测指标的调控作用、LPS则能够明显逆转HCQ对SLE小鼠各检测指标的调控作用(均P<0.05)。结论:HCQ可能通过抑制TLR4/NF-κB信号通路及其介导的炎性反应,从而减轻SLE小鼠肺损伤,改善其肺功能。Objective:To investigate the effect of hydroxychloroquine(HCQ)on lung injury and toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway in systemic lupus erythematosus(SLE)mice.Methods:32 female MRL/lpr lupus mice(spo ntaneous SLE mice model)were randomly divided into model group,HCQ(80 mg/kg)group,HCQ(80 mg/kg)+TAK242(TLR4 inhibitor,0.3 mg/kg)group,HCQ(80 mg/kg)+LPS(TLR4 agonist,2.5 mg/kg)group,with 8 mice in each group;another 8 female C57BL/6 mice of the same age were set as normal control group.After 5 weeks of continuous administration,the lung function indicators(airway resistance[Raw]),dynamic lung compliance[Cdyn]and forced expiratory volume at 0.1 second[FEV0.1]were detected by pulmonary function apparatus.The level of inflammatory factors(interferon-γ[IFN-γ]),tumor necrosis factor-α[TNF-α],(interleukin-1β[IL-1β]and IL-6)in alveolar lavage fluid and the immunological indexes(antinuclear antibody[ANA]and anti-double-stranded DNA[ds-DNA])in serum were detected by ELISA.The histopathological changes and fibrosis of lung tissue were observed by HE staining or Masson staining,and the pathological score and collagen volume fraction(CVF)were calculated.The mRNA and protein expressions of TLR4,NF-κB p65 and IκBαin lung tissues were detected by RT-PCR or Western blot.Results:Compared with the model group,the Raw of HCQ group was significantly decreased while the Cdyn,FEV0.1 were significantly increased(all P<0.05).The levels of IFN-γ,TNF-α,IL-1β,IL-6 in alveolar lavage fluid and the levels of ANA,ds-DNA in serum were significantly decreased(all P<0.05).The pathological changes and fibrosis of lung tissue were significantly improved,the pathological score and CVF were significantly decreased(all P<0.05).The mRNA and protein expressions of TLR4,NF-κB p65,IκBαin lung tissue were significantly decreased(all P<0.05).TAK242 could significantly enhance the regulation effect of HCQ on the detectied indexes in SLE mice(all P<0.05).LPS could significantly reverse the regulatory effects of HCQ

关 键 词：系统性红斑狼疮 羟氯喹 肺损伤 肺功能 TLR4/NF-κB信号通路 炎性反应 

分 类 号：R593.24[医药卫生—内科学]