保肺化纤方调控FGF2/FGFRs信号通路对BLM/PM_(2.5)诱导的IPF大鼠肺成纤维细胞分化的影响  

Effects of Baofei Huaxian Formula on differentiation of lung fibroblasts in IPF rat induced by BLM/PM_(2.5) via regulating FGF2/FGFRs signaling pathway

作　　者：林珊珊 马宁怡 何慧慧 贾瑞 千孟丹 王炎 黄慧敏 孔德琪 李亚[1,2,3] LIN Shanshan;MA Ningyi;HE Huihui;JIA Rui;QIAN Mengdan;WANG Yan;HUANG Huimin;KONG Deqi;LI Ya(Henan University of Chinese Medicine,Zhengzhou 450046,China;Respiratory Pharmacological Laboratory of Chinese Medicine,The First Affiliated Hospital of Henan University of CM,Zhengzhou 450000,China;Henan Key Laboratory of Chinese Medicine for Respiratory Disease,Co-construction Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan&Education Ministry of P.R.,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学,郑州450046 [2]河南中医药大学第一附属医院中药药理(呼吸)实验室,郑州450000 [3]河南省中医药防治呼吸病重点实验室,呼吸疾病中医药防治省部共建协同创新中心,郑州450046

出　　处：《中华中医药杂志》2023年第11期5220-5226,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.82074403);河南省中医药科学研究专项课题(No.20-21ZY1019);河南省特色骨干学科中医学学科建设项目(No.STG-ZYXKY-2020014);河南省重点研发与推广专项(科技攻关)(No.182102310102)。

摘　　要：目的:探讨保肺化纤方经成纤维细胞生长因子2(FGF2)/成纤维细胞生长因子受体(FGFRs)信号通路抑制博来霉素(BLM)/细颗粒物(PM_(2.5))诱导的肺成纤维细胞分化的作用机制。方法:以SD大鼠和人胚肺成纤维细胞(MRC-5)为研究对象,随机分为对照组、模型组、保肺化纤方组、尼达尼布组、保肺化纤方+尼达尼布组。采用一次性气管内雾化BLM联合大气实时浓缩PM_(2.5)暴露法诱导特发性肺纤维化(IPF)大鼠模型,采用BLM/PM_(2.5)联合诱导细胞模型,并给予相应药物干预。检测大鼠肺功能,采用HE、Masson染色观察肺组织病理和胶原沉积;采用免疫组织化学染色法检测肺组织α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(COLⅠ)、Ⅲ型胶原蛋白(COLⅢ)、FGF2、FGFR1、FGFR2、FGFR3、FGFR4蛋白表达水平;采用Western Blot法检测MRC-5细胞α-SMA、COLⅠ、COLⅢ、FGF2、FGFR1、FGFR2、FGFR3、FGFR4蛋白表达水平。结果:与对照组比较,模型组大鼠肺功能显著降低(P<0.01,P<0.05),肺组织及MRC-5细胞中α-SMA、COLⅠ、FGF2、FGFR1、FGFR2、FGFR3、FGFR4蛋白表达显著升高(P<0.01,P<0.05),COLⅢ在肺组织中表达显著升高(P<0.01);与模型组比较,保肺化纤方组大鼠肺功能显著升高(P<0.01,P<0.05),肺组织及MRC-5细胞中α-SMA、COLⅠ、COLⅢ、FGF2、FGFR1、FGFR2、FGFR3、FGFR4蛋白表达显著降低(P<0.01,P<0.05)。结论:保肺化纤方可有效改善BLM/PM_(2.5)诱导的IPF大鼠肺组织病理损伤,减少胶原沉积,其机制可能与下调FGF2/FGFRs信号通路抑制肺成纤维细胞分化有关。Objective:To explore the mechanism of Baofei Huaxian Formula(BHF)in inhibiting the differentiation of lung fibroblasts(MRC-5 cell line)induced by bleomycin(BLM)/PM_(2.5) via fibroblast growth factor 2(FGF2)/fibroblast growth factor receptors(FGFRs)signaling pathway.Methods:SD rats and MRC-5 cells were used as research objects,and they were randomly divided into control group,model group,Baofei Huaxian Formula(BHF),Nintedanib(Nint)and BHF+Nint groups.The IPF rat model was duplicated by one-time endotracheal nebulization of BLM combined with PM2.s exposure method,and the cell model was induced by BLM combined with PM2.s suspension,and administrated with corresponding drugs.The lung function of rats was detected.HE and Masson staining were used to observe the lung tissues pathology and collagen deposition.Immunohistochemistry was used to detectα-smooth muscle actin(α-SMA),type Ⅰ collagen(COL Ⅰ),type Ⅲ collagen(COLⅢ),FGF2,FGFR1,FGFR2,FGFR3 and FGFR4 protein expression levels in lung tissues.The protein expression levels of α-SMA,COL Ⅰ,COL Ⅲ,FGF2,FGFR1,FGFR2,FGFR3 and FGFR4 in MRC-5 cells were detected by Western Blot.Results:Compared with the Control group,the lung function significantly decreased(P<0.01,P<0.05),the expressions of α-SMA,COL Ⅰ,FGF2,FGFR1,FGFR2,FGFR3 and FGFR4 in the lung tissues and MRC-5 cells were significantly increased in the model group(P<0.01,P<0.05),the expression of COLⅢ in lung tssues was significantly increased(P<0.01).Compared with the model group,the lung function significantly increased(P<0.01,P<0.05),α-SMA,COL Ⅰ,COL Ⅲ,FGF2,FGFR1,FGFR2,FGFR3 and FGFR4 proteins in lung tissues and MRC-5 cells in BHF group were significantly decreased(P<0.01,P<0.05).Conclusion:Baofei Huaxian Formula can markedly improve the lung tissue pathological damage,and reduced collagen deposition,and the regulatory mechanisms of inhibiting lung fibroblast differentiation via down-regulated FGF2/FGFRs signaling pathway might be involved.

关 键 词：保肺化纤方 肺成纤维细胞分化 FGF2/FGFRs信号通路 细颗粒物 特发性肺纤维化 

分 类 号：R285.5[医药卫生—中药学]