柴胡皂甙-d调控ECM微环境抑制肺纤维化的机制研究  

Studies Related to the Regulation of the ECM Microenvironment of Pulmonary Fibrosis by Saikosaponin d

作　　者：徐云聪 郑金旭[1] XU Yuncong;ZHENG Jinxu(Department of Respiratory Medicine,Jiangsu University Affiliated Hospital,Zhenjiang 212000,China)

机构地区：[1]江苏大学附属医院呼吸内科,江苏镇江212000

出　　处：《药物生物技术》2025年第1期50-57,共8页Pharmaceutical Biotechnology

基　　金：镇江市重点研发计划(No.SH2018048);苏州市科技发展计划(No.SYSD2020010);江苏大学2023年度医教协同创新基金一般项目(No.JDYY2023034)。

摘　　要：探究柴胡皂甙-d(saikosaponin-d,SSd),对肺纤维化细胞外基质(extracellular matrix,ECM)的调控及作用机制。随机将C57BL/6小鼠分为四组(每组n=20):空白对照(control,Cont)组,博来霉素(bleomycin,BLM)组、柴胡皂甙-d(saikosaponin,SSd)组,地塞米松(dexamethasone,DXM)组。通过气管内分别注射BLM至BLM组、SSd组、DXM组小鼠肺组织中完成IPF模型的建立,并在SSd组和DXM组每日腹腔注射药物干预。在建模后第3、7、14、28 d,通过染色进行组织病理学检查。用酶联免疫吸附技术(enzyme-linked immunosorbent assay,ELISA)检测肺匀浆液的羟脯氨酸(hydroxyproline,HYP)、IL-4、IL-10、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量,Western Blot技术检测建模后肺组织促纤维化因子TGF-β、IGF-1的表达。免疫组化技术、Western Blot技术检测肺组织中E-钙黏蛋白(E-cadherin,E-cad)、AKT1、纤维连接蛋白-蛋白结构域B(fibronectin-extra domain B,FN-EDB)的表达。与Cont组相比,BLM组、SSd组、DXM组TGF-β、IGF-1、HYP、AKT1、FN-EDB、TNF-α、IL-4表达升高,E-cad表达降低;与BLM组相比,SSd组、DXM组TGF-β、IGF-1、HYP、AKT1、FN-EDB、TNF-α、IL-4表达降低,E-cad表达有所回升,IL-10表达差异不明显。肺纤维化进展中TGF-β、IGF-1升高,并受到SSd的抑制,SSd组通过调控PI3K/AKT通路缓解ECM的异常沉积和纤维化区域的炎症因子释放,改变ECM的微环境,延缓肺纤维化的进展。To investigate the regulation and mechanism of saikosaponin-d on the extracellular matrix(ECM)of pulmonary fibrosis,C57BL/6 mice were randomly allocated into four groups(n=20 in each group):blank control(Cont)group,bleomycin(bleomycin,BLM)group,saikosaponin-d(SSd)group,and dexamethasone(DXM)group.The IPF model was established by intratracheal injection of BLM into the lung tissues of mice in the BLM group,SSd group,and DXM group,and daily drug intervention was admi-nistered via intraperitoneal injection in the SSd group and DXM group.On the 3rd,7th,14th,and 28th days after modeling,histopathological examination was conducted through staining.The contents of hydroxyproline(HYP),IL-4,IL-10,and tumor necrosis factor-α(TNF-α)in lung homogenate were detected by enzyme-linked immunosorbent assay(ELISA),and the expression of profibrotic factors TGF-βand IGF-1 in lung tissue after modeling was determined by Western Blot technology.The expressions of E-cadherin(E-cad),AKT1,and fibronectin-extra domain B(FN-EDB)in lung tissue were examined by immunohistochemistry and Western blot technology.Compared with the Cont group,the expressions of TGF-β,IGF-1,HYP,AKT1,FN-EDB,TNF-α,and IL-4 in the BLM group,SSd group,and DXM group escalated,and the expression of E-cad declined.Compared with the BLM group,the expressions of TGF-β,IGF-1,HYP,AKT1,FN-EDB,TNF-α,and IL-4 in the SSd group and DXM group decreased,and the expression of E-cad recovered;the expression of IL-10 showed no significant difference.TGF-βand IGF-1 increased during the progression of pulmonary fibrosis and were inhibited by SSd.The SSd group mitigates the abnormal deposition of ECM and the release of inflammatory factors in the fibrotic area by regulating the PI3K/AKT pathway,alters the microenvironment of ECM,and delays the progression of pulmonary fibrosis.

关 键 词：特发性肺纤维化 柴胡皂甙-D 细胞外基质 胰岛素样生长因子-1 

分 类 号：R563.9[医药卫生—呼吸系统]