摘要
目的研究黄芪提取物(AME)对类风湿关节炎继发肺间质病变(RA-ILD)大鼠肺损伤及骨髓分化因子88(MyD88)/toll样受体4(TLR4)/核因子κB(NF-κB)p65通路的影响。方法将SD大鼠随机分为对照组、RA-ILD组、AME低剂量组(5 g/L)、AME高剂量组(10 g/L)、AME高剂量+脂多糖(LPS)组(10 g/L AME+1 mg/L TLR4激活剂LPS)。除对照组外,其余组大鼠均通过注射牛Ⅱ型胶原蛋白、弗氏完全佐剂和博莱霉素构建RA-ILD模型;检测大鼠关节炎指数、肺组织湿干重比;ELISA检测肺泡灌洗液炎症因子白细胞介素(IL)-1β、IL-6和肿瘤坏死因子-α(TNF-α)水平;苏木精-伊红染色观察大鼠膝关节组织和肺组织病理学变化;Western blot检测大鼠肺组织自噬因子苄氯素1(Beclin 1)、微管相关蛋白1A/1B-轻链3(LC3)Ⅱ/Ⅰ及MyD88/TLR4/NF-κB p65通路相关蛋白表达。结果与对照组比较,RA-ILD组大鼠膝关节组织和肺组织受损,关节炎指数、肺组织湿干重比、IL-1β、IL-6和TNF-α水平、MyD88、TLR4蛋白表达水平及p-NF-κB p65/NF-κB p65比值升高(P<0.01),Beclin 1、LC3Ⅱ/Ⅰ蛋白表达降低(P<0.01);与RA-ILD组比较,AME低剂量组、AME高剂量组大鼠组织损伤减轻,关节炎指数、肺组织湿干重比、IL-1β、IL-6和TNF-α水平、MyD88、TLR4蛋白表达水平及p-NF-κB p65/NF-κB p65比值呈剂量依赖性降低(P<0.05或P<0.01),Beclin 1、LC3Ⅱ/Ⅰ蛋白表达呈剂量依赖性升高(P<0.05或P<0.01);与AME高剂量组比较,AME高剂量+LPS组大鼠组织损伤加重,关节炎指数、肺组织湿干重比、IL-1β、IL-6和TNF-α水平、MyD88、TLR4蛋白表达水平及p-NF-κB p65/NF-κB p65比值升高(P<0.01),Beclin 1、LC3Ⅱ/Ⅰ蛋白表达降低(P<0.01)。结论AME抑制MyD88/TLR4/NF-κB p65通路,缓解RA-ILD大鼠肺损伤。
Objective To investigate the effects of astragalus mongholicus extract(AME)on lung injury and the myeloid differentiation factor 88(MyD88)/Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)p65 pathway in rheumatoid arthritis induced interstitial lung disease(RA-ILD)rats.Methods SD rats were randomly divided into a control group,RA-ILD group,low-dose AME group(5 g/L),high-dose AME group(10 g/L),and high-dose AME+lipopolysaccharide(LPS)group(10 g/L AME+1 mg/L TLR4 activator LPS).Except for the control group,rats in all other groups were injected with bovine typeⅡcollagen,Freund’s complete adjuvant,and bleomycin to establish the RA-ILD model.The arthritis index and lung tissue wet-dry weight ratio of rats were tested.ELISA was applied to detect the levels of inflammatory factors interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)in bronchoalveolar lavage fluid.Hematoxylin eosin staining was used to observe pathological changes of rat knee joint tissue and lung tissue.Western blot was applied to detect the expression of autophagy factors Beclin 1,microtubule-associated protein 1A/1B-light chain 3(LC3)Ⅱ/Ⅰ,and MyD88/TLR4/NF-κB p65 pathway related proteins in lung tissue.Results Compared with control group,knee joint tissue and lung tissue of rats in RA-ILD group were damaged,the arthritis index,lung tissue wet-dry weight ratio,levels of IL-1β,IL-6,and TNF-α,the expression levels of MyD88 and TLR4 proteins,and p-NF-κB p65/NF-κB p65 ratio increased(P<0.01),the expression of Beclin 1 and LC3Ⅱ/Ⅰproteins decreased(P<0.01).Compared with RA-ILD group,the low-dose and high-dose AME groups showed reduced tissue damage in rats,the arthritis index,lung tissue wet-dry weight ratio,levels of IL-1β,IL-6,and TNF-α,the expression levels of MyD88 and TLR4 proteins,and p-NF-κB p65/NF-κB p65 ratio showed a dose-dependent decrease(P<0.05 or P<0.01),the expression of Beclin 1 and LC3Ⅱ/Ⅰproteins showed a dose-dependent increase(P<0.05 or P<0.01).Compared with high-dose AME group,the tissue damage of rats in the high-dose AME+LPS group was worsened,the arthritis index,lung tissue wet-dry weight ratio,levels of IL-1β,IL-6,and TNF-α,the expression levels of MyD88 and TLR4 proteins,and p-NF-κB p65/NF-κB p65 ratio were higher(P<0.01),the expression of Beclin 1 and LC3Ⅱ/Ⅰproteins was lower(P<0.01).Conclusion AME inhibits the MyD88/TLR4/NF-κB p65 pathway and alleviates lung injury in RA-ILD rats.
作者
赵悦
杨金良
罗寰
席文秀
王珺璐
郑学军
Zhao Yue;Yang Jinliang;Luo Huan;Xi Wenxiu;Wang Junlu;Zheng Xuejun(Dept of Rheumatology and Immunology,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075000)
出处
《安徽医科大学学报》
北大核心
2025年第7期1173-1179,共7页
Acta Universitatis Medicinalis Anhui
基金
河北省医学科学研究课题计划项目(编号:20231416)。
