摘要
目的对金银花石油醚部位提取物抗肺纤维化药效作用进行探究,并对其药效成分进行分析。方法体外实验采用5 ng·mL^(-1)转化生长因子(TGF)-β1诱导A549细胞建立上皮间质转化(epithelial mesenchymal transition,EMT)细胞模型,设置空白对照组、模型组、金银花石油醚部位组,RT-PCR法检测药物干预后A549细胞EMT标志性基因(ColⅠ、Vimentin、E-cadherin、α-SMA)mRNA表达水平,ELISA法检测羟脯氨酸(HYP)含量变化。将36只雄性C57BL/6小鼠随机分为假手术组、模型组、吡非尼酮组和金银花石油醚部位组,暴露式气管内注入博来霉素5 mg·kg^(-1)进行模型复制,第7天开始给药,第28天处死小鼠。HE、Masson染色观察小鼠肺组织病理形态变化,ELISA法检测肺组织HYP含量,RT-PCR法检测肺组织EMT标志性基因表达水平。并对金银花石油醚部位进行UPLC-Q-Extractive/MS成分定性分析。结果与模型组比较,金银花石油醚部位可有效抑制TGF-β1诱导后A549细胞内ColⅠ、α-SMA、Vimentin(P<0.05,P<0.01)mRNA表达,促进E-cadherin mRNA(P<0.01,P<0.05)表达,降低细胞内羟脯氨酸(P<0.05,P<0.01)含量;金银花石油醚部位可改善小鼠肺组织炎症与纤维化程度,使肺组织ColⅠ、α-SMA、Vimentin mRNA(P<0.05,P<0.01)的表达量下降,E-cadherin mRNA表达量上升(P<0.05,P<0.01),羟脯氨酸含量下降(P<0.05,P<0.01)。LC-MS共鉴定出金银花石油醚部位中14种化学成分,以酚类和黄酮类成分为主。结论金银花石油醚部位通过抑制上皮间质转化过程产生抗肺纤维化作用。酚类和黄酮类成分是其潜在药效物质基础。
Objective To study the anti-pulmonary fibrosis effect of the petroleum ether extract of Lonicerae japonicae(PLJ)and predict its potential pharmacodynamic components.Methods An in vitro EMT cell model was established in A549 cells incubated with 5 ng·mL^(-1)of TGF-β1,blank control group,model group and petroleum ether part of Lonicera japonica groups were set up.RT-PCR was applied to detect mRNA expression level of EMT related genes(Col I,Vimentin,E-cadherin andα-SMA)in A549 after drug intervention;ELISA was used to detect the content of HYP.36 C57BL/6 male mice were randomly divided into sham operation group,bleomycin model group,pirfenidone group and petroleum ether part of Lonicera japonica groups at different doses,5 mg·mL^(-1)of bleomycin was intratracheally injected to establish the pulmonary fibrosis mice model.7 days later,pirfenidone and petroleum ether part of Lonicera japonica were daily orally administered,and all mice were sacrificed on the 28th day.The pathology of lung tissues was observed by HE and Masson staining,ELISA was used to detect HYP in lung tissues,and the expression of EMT related genes was detected by RT-PCR.The components of petroleum ether extract of Lonicerae japonica were qualitatively analyzed by the UPLC-Q-extractive/MS.Results Compared with the model group,petroleum ether part of Lonicera japonica can effectively inhibit the expression of ColⅠ,α-SMA and Vimentin(P<0.05,P<0.01)mRNA in TGF-β1-induced A549,promote the expression of E-cadherin mRNA(P<0.01,P<0.05),reduce the content of HYP(P<0.05,P<0.01).In addition,petroleum ether part of Lonicera japonica can improve the degree of pulmonary inflammation and fibrosis in mice,decrease the expressions of ColⅠ,α-SMA and Vimentin(P<0.05,P<0.01)mRNA,increase the expression of E-cadherin mRNA(P<0.05,P<0.01),and decrease the content of HYP(P<0.05,P<0.01).LC-MS results showed that a total of 14 chemical constituents were identified from the PLJ,among which phenols and flavonoids were the main constituents.Conclusion PLJ has anti-pulmonary fibrosis effect via inhibiting the process of EMT,phenols and flavonoids are potential medicinal components.
作者
李雅群
杨勇
王旭
王萍
田景振
朱庆军
容蓉
LI Yaqun;YANG Yong;WANG Xu;WANG Ping;TIAN Jingzhen;ZHU Qingjun;RONG Rong(Shandong University of Traditional Chinese Medicine,Jinan 250355 Shandong,China;Huashan Hospital of Fudan University,Shanghai 200000,China;Antiviral Synergistic Innovation Center of Traditional Chinese Medicine in Universities of Shandong Province,Jinan 250355 Shandong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2021年第9期1268-1275,共8页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金面上项目(81873220,81774167)
山东省重点研发计划项目(重大科技创新工程,2020CXGC010505)。
