绿原酸灌胃对博莱霉素诱导小鼠肺纤维化的改善作用及其机制  

Ameliorative effect of intragastric administration of chlorogenic acid on bleomycin⁃induced pulmonary fibrosis in mice and its mechanism

作　　者：王倩 刘宇 王荣丽 WANG Qian;LIU Yu;WANG Rongli(Department of Respiratory and Critical Care Medicine,The Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学附属医院呼吸与危重症医学科,四川泸州646000

出　　处：《山东医药》2023年第18期44-49,共6页Shandong Medical Journal

摘　　要：目的观察绿原酸(CGA)对博莱霉素诱导的小鼠肺纤维化的改善作用,并探讨其可能的作用机制。方法将60只雄性C57BL/6小鼠随机分为CGA低剂量组、CGA中剂量组、CGA高剂量一组、CGA高剂量二组、模型组及对照组,每组各10只。除对照组外其余5组小鼠均气管内一次性注入博莱霉素2 mg/kg(记作第1天),第1天小鼠自然苏醒后,CGA高剂量二组小鼠继续予CGA 80 mg/kg灌胃,1次/天,连续灌胃14天;第8天时CGA低剂量组、CGA中剂量组、CGA高剂量一组分别予绿原酸20、40、80 mg/kg灌胃,1次/天,连续灌胃14天;第8天时对照组及模型组予等体积超纯水连续灌胃,1次/天,连续灌胃14天。第21天后处死各组小鼠取肺组织,观察各组小鼠肺组织肺纤维化程度(肺系数、HE染色观察肺组织形态学、Masson染色观察肺组织胶原纤维沉积及Ashcroft评分);采用免疫荧光法检测各组肺组织α⁃平滑肌肌动蛋白(α⁃SMA)表达,采用免疫组化法检测各组肺组织胶原蛋白Ⅰ(CollagenⅠ)、波形蛋白(Vimentin)、转化生长因子⁃β1(TGF⁃β1)、Smad2/3蛋白表达,采用TBA法检测各组肺组织丙二醛(MDA)含量,采用RT⁃qPCR法检测各组肺组织E⁃钙连蛋白(E⁃cadherin)及α⁃SMA mRNA。结果与对照组相比,模型组小鼠肺组织正常结构遭到严重破坏,胶原纤维大量沉积,肺系数及Ashcroft评分高(P<0.001);与模型组相比,CGA各剂量组小鼠肺组织结构破坏程度减轻,胶原纤维沉积减少,肺系数及Ashcroft评分低(P均<0.001),且呈剂量依赖性(P<0.001);与对照组比较,模型组小鼠肺组织中MDA含量高,肺组织E⁃cadherin mRNA相对表达量低、α⁃SMA mRNA相对表达量高,肺组织α⁃SMA、Col⁃lagenⅠ、Vimentin、TGF-β1、Smads2/3蛋白相对表达量高(P均<0.001);与模型组比较,CGA各剂量组随作用浓度升高,各组小鼠肺组织α⁃SMA、CollagenⅠ、Vimentin、TGF-β1、Smads2/3蛋白相对表达量降低(P均<0.01),α⁃SMA mRNA相对表达量Objective To observe the therapeutic effect of chlorogenic acid(CGA)on bleomycin⁃induced pulmo⁃nary fibrosis in mice,and to explore its possible mechanism.Methods Sixty male C57BL/6 mice were randomly divid⁃ed into the low⁃dose CGA group,medium⁃dose CGA group,high⁃dose CGA group 1,high⁃dose CGA group 2,model group,and control group,with 10 mice in each group.Except the control group,mice in the other 5 groups were injected with bleomycin 2 mg/kg into the trachea at once(labeled as day 1).After the mice recovered naturally on day 1,mice in the high⁃dose CGA group 2 were continued to receive CGA 80 mg/kg through intragastric administration,once a day,for 14 days.On day 8,mice in the low⁃dose CGA group,medium⁃dose CGA group and high⁃dose CGA group 1 were given 20,40 and 80 mg/kg chlorogenic acid by gavage,once a day,for 14 days.At the same time,on the 8th day,mice in the control group and the model group were given the same volume of ultra⁃pure water by gavage,once a day,for 14 days.Af⁃ter 21 days,the mice in each group were sacrificed,and the lung tissues were taken to observe the degree of pulmonary fi⁃brosis.The lung coefficient was calculated.HE staining was used to observe the morphological changes of the lung tissues under light microscope.Masson staining was used to observe the collagen fiber deposition,and Ashcroft score was used to evaluate the degree of pulmonary fibrosis.The immunofluorescence method was used to detect the expression ofα⁃smooth muscle actin(α⁃SMA)in the lung tissues.The protein expression levels of Collagen I,Vimentin,transform⁃ing growth factor⁃β1(TGF⁃β1),and Smad2/3 in the lung tissues were detected by immunohistochemistry.The content of malondialdehyde(MDA)in lung tissue was determined by TBA method.The mRNA expression levels of E⁃cadherin andα⁃SMA in the lung tissues were detected by real-time qPCR.Results Compared with the control group,the struc⁃ture of the lung tissues in the model group was seriously damaged,collagen fibers were dep

关 键 词：绿原酸 肺纤维化 特发性肺纤维化 博莱霉素 上皮—间充质转化 TGF⁃β1/Smads信号通路 

分 类 号：R563.1[医药卫生—呼吸系统]