摘要
目的 探讨养阴清肺汤通过对Toll样受体4(TLR4)信号和核因子κB受体激活剂配体(RANKL)信号的调控,发挥抑制矽肺小鼠肺纤维化的作用机制.方法 将72只SPF级C57BL/6J雄性小鼠随机分为对照组、矽肺组、吡非尼酮组和养阴清肺汤高、中、低剂量组,每组12只.采用气管一次性灌注5mg SiO2建立矽肺小鼠模型,2周后予以200mg/kg/d吡非尼酮,1 000、500、250mg/kg/d高、中、低剂量养阴清肺汤连续灌胃2周.HE染色、Masson染色观察肺组织病理形态和胶原沉积.微小计算机断层扫描(micro-CT)重建肺三维结构;全身体积描记系统(WBP)检测肺功能;免疫荧光染色标记Toll样受体4(TLR4);抗酒石酸酸性磷酸酶(TRAP)染色标记肺组织阳性细胞;免疫印迹法检测肺组织Ⅰ型胶原蛋白(CoL Ⅰ)、波形蛋白(Vimen-tin)、Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核因子κB(NF-κB)、白细胞介素6(IL-6)、核因子κB受体激活因子(RANK)、核因子κB受体激活因子配体(RANKL)的蛋白表达.结果 与矽肺模型组比较,养阴清肺汤可减少矽结节面积和胶原沉积(P＜0.05),改善矽肺小鼠肺功能(P＜0.05),养阴清肺汤组ColI、vimentin、TLR4、MyD88、NF-κB、IL-6、RANK 和 RANKL 蛋白水平均明显下调(P＜0.05).结论 养阴清肺汤可通过抑制TLR4和RANKL信号的活化,改善肺功能和减轻矽肺小鼠肺纤维化进展.
Abstract
Objective To explore the effects and mechanisms of Yang-Yin-Qing-Fei Decoction(YYQFD)on lung fibrosis in silicotic mice by regulating Toll-like receptors 4(TLR4)and Receptor activator of nuclear fac-tor kappa-B ligand(RANKL)signal pathways.Methods Seventy-two SPF grade C57BL/6J male mice were randomly divided into six groups(n=12),as control group,silicosis group,pirfenidone treatment group,and YYQFD high-dose,medium-dose,low-dose treatment groups.Silicotic mice was made by intratracheal instillation of SiO2(5mg/mice)for 2 weeks and treated with YYQFD(1 000 mg,500 mg,250mg/kg/d)or pirfenidone(200mg/kg/d)for another 2 weeks.The pathological morphology and collagen deposition of lung tissue were observed HE staining and Masson staining.Three-dimensional lung structure was reconstructed by micro computed tomography(Micro-CT).Whole body plethysmography(WBP)was used to detect lung functions.The expression of TLR4 in lung tissue was was observed by immunofluorescence staining.Tar-trate-resistant acid phosphatase(TRAP)staining was used to observe the expression of positive cells in lung tissue.The expression of Type Ⅰ collagen(Col Ⅰ),Vimentin,TLR4;Myeloid differentiation primary re-sponse gene 88(MyD88),Nuclear factor kappa-B(NF-κB),interleukin 6(IL-6),Receptor activator of nu-clear factor kappa-B ligand(RANK),and RANKL in lung tissue was detected by Western blot.Results Compared with the silicosis group,YYQFD treatment groups attenuated silicotic area,collagen deposition and improved impair lung functions in silicotic mice(P＜0.05).The expression of Col Ⅰ,vimentin,TLR4,MyD88,NF-κB,IL-6,RANK and RANKL were all reduced in YYQFD groups(P＜0.05).Conclusion YYQFD alleviated lung fibrosis and improved lung function in mice exposed to silica by regulating TLR4 and RANKL signal pathways.
基金项目
国家自然科学基金(82174212)
河北省自然科学基金(H2022209073)
河北省高等学校科学技术研究项目(ZD2021086)
维普
万方数据