中药新药与临床药理2026，Vol.37Issue(1)：31-41,11.DOI:10.19378/j.issn.1003-9783.2026.01.004
康复新液对脂多糖诱导炎症微环境中人食管成纤维细胞损伤的作用及机制
Effects and Mechanism of Kangfuxin Liquid on Human Esophageal Fibroblast Injury in a LPS-Induced Inflammatory Microenvironment
摘要
Abstract
Objective To investigate the effects and underlying mechanism of Kangfuxin Liquid[an extract of Periplaneta americana(L.)]on human esophageal fibroblast(HEFs)injury within a lipopolysaccharide(LPS)-induced inflammatory microenvironment.Methods(1)Following intervention with varying concentrations of Kangfuxin Liquid(0%,0.5%,1%,2%,4%,8%,16%,32%)and LPS(0,2,4,6,8,10,20,40 μg·mL-1)for 24 hours,the HEFs proliferation rate was assessed via the CCK-8 assay to determine the optimal concentrations for LPS induction and Kangfuxin Liquid intervention.(2)HEFs in the logarithmic growth phase were divided into the following groups:Control,LPS,0.5%Kangfuxin Liquid,1%Kangfuxin Liquid,2%Kangfuxin Liquid,and Prednisolone Acetate(PDNN,50 μg·mL-1).An inflammatory microenvironment was induced by stimulation with LPS(10 μg·mL-1)for 1 hour.The levels of interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,and inducible nitric oxide synthase(iNOS)in the cell supernatant were measured by ELISA.The expression levels of IL-6,nuclear factor(NF)-κB p-p65,p-Smad3,and α-smooth muscle actin(α-SMA)in the cells were detected by immunofluorescence.The mRNA expression levels of IL-1β,IL-6,TNF-α,NF-κB,transforming growth factor(TGF)-β1,Smad3,and α-SMA in the cells were determined using qRT-PCR.The protein expression levels of NF-κB p65,NF-κB p-p65,inhibitor of nuclear factor kappa-B alpha(IκBα),p-IκBα,TGF-β1,Smad3,p-Smad3,and α-SMA in the cells were assessed by Western Blot.Results(1)Subsequent experiments utilized Kangfuxin Liquid at concentrations of 0.5%,1%,and 2%,and an LPS concentration of 10 μ g·mL-1 to induce the inflammatory microenvironment in HEFs.(2)Compared with the control group,the LPS group exhibited significantly increased levels of IL-1β,IL-6,TNF-α,and iNOS in the HEF supernatant(P＜0.01);significantly upregulated mRNA expression of IL-1β,IL-6,TNF-α,and NF-κB(P＜0.01);significantly increased protein expression of NF-κB p65,NF-κB p-p65,IκBα,and p-IκBα(P＜0.01);significantly enhanced cytoplasmic red fluorescence signal for IL-6 and nuclear green fluorescence signal for NF-κB p-p65(P＜0.01);significantly upregulated mRNA expression of TGF-β1,Smad3,and α-SMA(P＜0.01);significantly increased protein expression of TGF-β1,Smad3,p-Smad3,and α-SMA(P＜0.01);and significantly intensified nuclear green fluorescence signal for p-Smad3 and cytoplasmic red fluorescence signal for α-SMA(P＜0.01).Compared with the LPS group,the 0.5%,1%,and 2%Kangfuxin Liquid groups and the PDNN group showed significantly reduced levels of IL-6,TNF-α,and iNOS in the cell supernatant(P＜0.01);significantly downregulated protein expression of NF-κB p65,NF-κB p-p65,IκBα,and p-IκBα(P＜0.01);significantly attenuated cytoplasmic red fluorescence signal for IL-6 and nuclear green fluorescence signal for NF-κB p-p65(P＜0.01);significantly decreased protein expression of TGF-β1 and p-Smad3(P＜0.01);and significantly weakened cytoplasmic red fluorescence signal for α-SMA(P＜0.01).Compared with the LPS group,the 2%Kangfuxin Liquid group and the PDNN group demonstrated significantly lower IL-1β content in the cell supernatant(P＜0.01);significantly downregulated mRNA expression of IL-1β,IL-6,TNF-α,and NF-κB(P＜0.05,P＜0.01);significantly downregulated mRNA expression of TGF-β1,Smad3,and α-SMA(P＜0.01);significantly decreased protein expression of Smad3 and α-SMA(P＜0.01);and a significantly weakened nuclear green fluorescence signal for p-Smad3(P＜0.01).Conclusion Kangfuxin Liquid can alleviate HEF injury and inhibit the phenotypic transition to myofibroblasts in an LPS-induced inflammatory microenvironment.The underlying mechanism may be associated with the suppression of the excessive activation of the TGF-β1/Smad3 signaling pathway.These findings provide experimental evidence supporting the potential use of Kangfuxin Liquid in treating esophageal strictures following endoscopic submucosal dissection.关键词
康复新液/美洲大蠊提取物/食管狭窄/炎症微环境/人食管成纤维细胞/炎症反应/纤维化/TGF-β1/Smad3 信号通路Key words
Kangfuxin Liquid/Periplaneta americana extract/esophageal stricture/inflammatory microenvironment/human esophageal fibroblasts/inflammatory response/fibrosis/TGF-β1/Smad3 signaling pathway分类
医药卫生引用本文复制引用
周鑫,赵龙,王洪连,罗钰婷,李志,刘建琴..康复新液对脂多糖诱导炎症微环境中人食管成纤维细胞损伤的作用及机制[J].中药新药与临床药理,2026,37(1):31-41,11.基金项目
四川省科技厅重点研发项目(2020YFS0376) （2020YFS0376）
西南医科大学科研创新团队项目(2022-CXTD-01). （2022-CXTD-01）
