基于主成分分析研究青翘、老翘对博来霉素诱导肺纤维化小鼠的作用差异  

Differential effects of Qingqiao and Laoqiao on bleomycin-induced pulmonary fibrosis in mice by principal component analysis

作　　者：张钦钦 张贝贝 杨帆[1] 王茜 薛淑娟 布晨希 吴给给 陈随清[1,2,3] ZHANG Qin-qin;ZHANG Bei-bei;YANG Fan;WANG Xi;XUE Shu-juan;BU Chen-xi;WU Gei-gei;CHEN Sui-qing(Henan University of Chinese Medicine,Zhengzhou 450046,China;Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province and Education Ministry of China,Henan University of Chinese Medicine,Zhengzhou 450046,China;Collaborative Innovation Center for Whole-Chain Research and Development of Yu Medicine,Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学,河南郑州450046 [2]河南中医药大学呼吸疾病中医药防治省部共建协同创新中心,河南郑州450046 [3]河南中医药大学豫药全产业链研发河南省协同创新中心,河南郑州450046

出　　处：《中国中药杂志》2024年第15期4128-4138,共11页China Journal of Chinese Materia Medica

基　　金：现代农业产业技术体系建设——中药材首席专家项目(14104723-2022);河南省重大科技专项(221100310400)。

摘　　要：从药效相关性和成分差异阐释青翘与老翘改善博来霉素(BLM)诱导肺纤维化小鼠的作用差异。小鼠随机分为正常组,模型组,吡非尼酮组(50 mg·kg^(-1)),青翘低、高剂量组(1.3、2.6 g·kg^(-1)),老翘低、高剂量组(1.3、2.6 g·kg^(-1))。采用气管滴注BLM制备肺纤维化模型,统计小鼠存活率及体质量变化;给药完成后计算肺指数,HE染色、Masson染色和天狼星红染色评价肺组织病理变化;透射电子显微镜观察肺组织超微结构;生化法测定肺组织转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)、E钙黏着蛋白(E-cadherin)、羟脯氨酸(HYP)及肺泡灌洗液中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;RT-qPCR、Western blot法检测肺组织中基质金属蛋白酶7(MMP7)、胶原蛋白Ⅰ型(collagenⅠ)、E-cadherin、TNF-α、波形蛋白(vimentin)、TGF-β1、α-SMA表达;免疫荧光检测肺组织中α-SMA的表达;主成分分析法评价老翘与青翘改善肺纤维化的作用差异;分子对接技术分析老翘中含量高的化合物与TGF-β1受体之间的相互作用,CCK-8法检测其对TGF-β1诱导BEAS-2B和HFL1细胞模型的影响。结果显示,青翘、老翘可提高肺纤维化小鼠存活率,降低肺指数,改善肺组织病理损伤与胶原纤维沉积水平,改善小鼠Ⅱ型肺泡上皮细胞板层小体受损情况,降低肺泡灌洗液中IL-6、TNF-α水平,下调肺组织中HYP、MMP7、vi-mentin、collagenⅠ、TGF-β1、α-SMA的表达,上调E-cadherin的表达。通过主成分分析综合评价胶原纤维沉积水平,其改善程度为老翘高剂量>老翘低剂量>青翘高剂量>青翘低剂量。分子对接显示羟基酪醇、咖啡酸、连翘脂素、落叶松脂素与TGF-β1受体有较好的结合活性,且可显著减轻TGF-β1诱导BEAS-2B细胞损伤,抑制TGF-β1诱导HFL1细胞增殖。综上,青翘、老翘均可有效改善BLM诱导的肺纤维化小鼠损伤,老翘在减少胶原沉积方面优于青翘,可能与老翘中�The mechanism of alleviating bleomycin-induced pulmonary fibrosis in mice was compared between Qingqiao(Forsythiae Fructus produced with immature fruits)and Laoqiao(Forsythiae Fructus produced with mature fruits)from the pharmacodynamic correlation and composition differences.Mice were randomized into normal,model,pirfenidone(50 mg·kg^(-1)),low-and high-dose(1.3,2.6 g·kg^(-1),respectively)Qingqiao,and low-and high-dose(1.3,2.6 g·kg^(-1),respectively)Laoqiao groups.The mouse model of pulmonary fibrosis was established by intratracheal instillation of bleomycin,during which the survival rate and body weight changes of the mice were measured.After modeling,the lung index was calculated,and the pathological changes in the lung tissue were evaluated by hematoxylin-eosin(HE),Masson,and Sirius red staining.Transmission electron microscopy was employed to observe the ultrastructure of the lung tissue.The biochemical assay was employed to measure the levels of transforming growth factor-β1(TGF-β1),α-smooth muscle actin(α-SMA),E-cadherin,and hydroxyproline(HYP)in the lung tissue and interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the bronchoalveolar lavage fluid(BALF).The mRNA and protein levels of matrix metalloproteinase 7(MMP7),collagenⅠ,E-cadherin,TNF-α,vimentin,TGF-β1,andα-SMA in the lung tissue were determined by RT-qPCR and Western blot,respectively.The expression ofα-SMA in the lung tissue was detected by the immunofluorescence assay.Principal component analysis was performed to compare the effects of Qingqiao and Laoqiao in ameliorating pulmonary fibrosis.Molecular docking was employed to analyze the binding between the compounds with high content in Laoqiao and TGF-β1.The cell-counting kit(CCK-8)assay was used to examine the effects of the active compounds on TGF-β1-induced BEAS-2B and HFL1 cell models.The results showed that Qingqiao and Laoqiao increased the survival rate,reduced the lung index,alleviated the pathological damage and collagen deposition in the lung tissue,ameliorated the

关 键 词：老翘 青翘 肺纤维化 主成分分析 

分 类 号：R285.5[医药卫生—中药学]