OBJECTIVE:To explore the underlying mechanisms of the effects of Yangqing Chenfei formula(养清尘肺方,YCF) on inflammation and fibrosis in silicosis via inhibition of macrophage polarization.METHODS:A silicotic rat mod...OBJECTIVE:To explore the underlying mechanisms of the effects of Yangqing Chenfei formula(养清尘肺方,YCF) on inflammation and fibrosis in silicosis via inhibition of macrophage polarization.METHODS:A silicotic rat model was established via a single intratracheal instillation of silica particles on the first day of week 0.Subsequently,YCF was administered intragastrically to silicotic rats during weeks 0-2 and 5-8 twice daily.The mouse-derived alveolar macrophage cell line was used to investigate the mechanisms of YCF in M1/M2 polarization.RESULTS:YCF treatment effectively inhibited lung pathological changes,including inflammatory cell infiltration and tissue damage,and increased the forced expiratory volume in the first 0.3 s,functional residual capacity,and maximal mid-expiratory flow in weeks 2 and 8.Furthermore,the treatment improved lung functions by upregulating tidal volume,pause increase,and expiratory flow at 50% tidal volume from weeks 5 to 8.Moreover,YCF could significantly suppressed the progression of inflammation and fibrosis,by reducing the levels of inflammatory cytokines,as well as collagen-Ⅰ and Ⅲ.YCF treatment also decreased the numbers of macrophages and M1/M2 macrophages and the level of transforming growth factor-β(TGF-β).Additionally,YCF5,the effective substance in YCF,decreased lipopolysaccharide and interferon-γ-induced M1 macrophage polarization in a concentration-dependent manner.The mechanism of anti-M1 polarization might be related to a decrease in extracellular signal-regulated kinase,c-JUN N-terminal kinase,P38,and P65 phosphorylation.Furthermore,YCF5 inhibited interleukin-4-induced M2 macrophages by decreasing the protein and m RNA expressions of arginase-1 and CD206 as well as the levels of profibrotic factors,such as TGF-β and connective tissue growth factor.The mechanisms underlying the anti-M2 polarization of YCF5 were primarily associated with the inhibition of the nuclear translocation of phosphorylated signal transducer and activator of transcription 6(pSTAT6).CONCLUSION:YCF significantly inhibits inflammation and fibrosis in silicotic rats probably via the suppression of M1/M2 macrophage polarization mediated by the inhibition of mitogen-activated protein kinase and nuclear factor kappa B signaling pathways and Janus kinase/STAT6 pathways.展开更多
OBJECTIVE:To explore the mechanisms of Yangqing Chenfei formula(养清尘肺方,YCF)in the treatment of silicosis through a comprehensive strategy consisting of serum pharmacochemistry,network pharmacology analysis,and in ...OBJECTIVE:To explore the mechanisms of Yangqing Chenfei formula(养清尘肺方,YCF)in the treatment of silicosis through a comprehensive strategy consisting of serum pharmacochemistry,network pharmacology analysis,and in vitro validation.METHODS:An ultrahigh-performance liquid chromatography-tandem mass spectrometry method was used to confirm the active components in YCF-medicated serum.Then,we obtained targets for active components and genes for silicosis from multiple databases.Furthermore,a protein-protein interaction network was constructed,and Kyoto Encyclopedia of Genes and Genomes pathway and biological process analyses were conducted to elucidate the mechanisms of YCF for the treatment of silicosis.Finally,we validated the important components and mechanisms in vitro.RESULTS:Altogether,19 active components were identified from rat serum after YCF administration.We identified 724 targets for 19 components,which were mainly related to inflammation[phosphatidy linositol 3 kinase/protein kinase B,forkhead box O,hypoxia inducible factor,and T-cell receptor signaling pathway,nitric oxide biosynthetic process],fibrotic processes[vascular endothelial growth factor signaling pathway,extracellular signal regulated kinase(ERK)1 and ERK2 cascade,smooth muscle cell proliferation],and apoptosis(negative regulation of apoptotic process).In addition,218 genes for silicosis were identified and were mainly associated with the inflammatory response and immune process[cytokine-cytokine receptor interaction,tumor necrosis factor alpha(TNF-α),toll-like receptor,and nucleotide binding oligomerization domain-like receptor signaling pathway].Taking an intersection of active component targets and silicosis genes,we obtained 61 common genes that were mainly related to the inflammatory response and apoptosis,such as the phosphatidylinositol-3-kinase/protein kinase B signaling pathway,mitogen activated protein kinases signaling pathway,TNF signaling pathway,toll-like receptor signaling pathway,biosynthesis of nitric oxide,and apoptotic process.In the herb-component-gene-pathway network,paeoniflorin,rutin and nobiletin targeted the most genes.In vitro,paeoniflorin,rutin and nobiletin decreased the mRNA levels of inflammatory factors[interleukin(IL)-6,TNF-α,and IL-1β],suppressed p-AKT and cleaved caspase-3,and increased B cell lymphoma(Bcl)-2 protein expression in silica-induced macrophages in a concentration-dependent manner.CONCLUSION:YCF could significantly relieve the inflammatory response of silicosis via suppression of the AKT/Bcl-2/Caspase-3 pathway.展开更多
基金Special Project of Traditional Chinese Medicine Research of Henan Province(2021ZYZD01)Evaluation of the Therapeutic Effect and Characteristics of Traditional Chinese Medicine Dialectical Treatment for Coal Worker's Pneumoconiosis Based on A Multicenter,Randomized,Double-Blind,Parallel Controlled Trial+1 种基金National Natural Science Fund of China(81973822)Exploring the Mechanism of Bufei Yishen Formula Inhibiting Inflammatory Response in the Treatment of COPD Based on Group Allocation Theory。
文摘OBJECTIVE:To explore the underlying mechanisms of the effects of Yangqing Chenfei formula(养清尘肺方,YCF) on inflammation and fibrosis in silicosis via inhibition of macrophage polarization.METHODS:A silicotic rat model was established via a single intratracheal instillation of silica particles on the first day of week 0.Subsequently,YCF was administered intragastrically to silicotic rats during weeks 0-2 and 5-8 twice daily.The mouse-derived alveolar macrophage cell line was used to investigate the mechanisms of YCF in M1/M2 polarization.RESULTS:YCF treatment effectively inhibited lung pathological changes,including inflammatory cell infiltration and tissue damage,and increased the forced expiratory volume in the first 0.3 s,functional residual capacity,and maximal mid-expiratory flow in weeks 2 and 8.Furthermore,the treatment improved lung functions by upregulating tidal volume,pause increase,and expiratory flow at 50% tidal volume from weeks 5 to 8.Moreover,YCF could significantly suppressed the progression of inflammation and fibrosis,by reducing the levels of inflammatory cytokines,as well as collagen-Ⅰ and Ⅲ.YCF treatment also decreased the numbers of macrophages and M1/M2 macrophages and the level of transforming growth factor-β(TGF-β).Additionally,YCF5,the effective substance in YCF,decreased lipopolysaccharide and interferon-γ-induced M1 macrophage polarization in a concentration-dependent manner.The mechanism of anti-M1 polarization might be related to a decrease in extracellular signal-regulated kinase,c-JUN N-terminal kinase,P38,and P65 phosphorylation.Furthermore,YCF5 inhibited interleukin-4-induced M2 macrophages by decreasing the protein and m RNA expressions of arginase-1 and CD206 as well as the levels of profibrotic factors,such as TGF-β and connective tissue growth factor.The mechanisms underlying the anti-M2 polarization of YCF5 were primarily associated with the inhibition of the nuclear translocation of phosphorylated signal transducer and activator of transcription 6(pSTAT6).CONCLUSION:YCF significantly inhibits inflammation and fibrosis in silicotic rats probably via the suppression of M1/M2 macrophage polarization mediated by the inhibition of mitogen-activated protein kinase and nuclear factor kappa B signaling pathways and Janus kinase/STAT6 pathways.
基金Special Project of Traditional Chinese Medicine Research in Henan Province:Evaluation and Characteristics of the Therapeutic Effect of Dialectical Treatment for Coal Worker Pneumoconiosis Based on Multicenter,Randomized,Double-Blind,and Parallel Controlled Trials(No.20-21ZYZD01)。
文摘OBJECTIVE:To explore the mechanisms of Yangqing Chenfei formula(养清尘肺方,YCF)in the treatment of silicosis through a comprehensive strategy consisting of serum pharmacochemistry,network pharmacology analysis,and in vitro validation.METHODS:An ultrahigh-performance liquid chromatography-tandem mass spectrometry method was used to confirm the active components in YCF-medicated serum.Then,we obtained targets for active components and genes for silicosis from multiple databases.Furthermore,a protein-protein interaction network was constructed,and Kyoto Encyclopedia of Genes and Genomes pathway and biological process analyses were conducted to elucidate the mechanisms of YCF for the treatment of silicosis.Finally,we validated the important components and mechanisms in vitro.RESULTS:Altogether,19 active components were identified from rat serum after YCF administration.We identified 724 targets for 19 components,which were mainly related to inflammation[phosphatidy linositol 3 kinase/protein kinase B,forkhead box O,hypoxia inducible factor,and T-cell receptor signaling pathway,nitric oxide biosynthetic process],fibrotic processes[vascular endothelial growth factor signaling pathway,extracellular signal regulated kinase(ERK)1 and ERK2 cascade,smooth muscle cell proliferation],and apoptosis(negative regulation of apoptotic process).In addition,218 genes for silicosis were identified and were mainly associated with the inflammatory response and immune process[cytokine-cytokine receptor interaction,tumor necrosis factor alpha(TNF-α),toll-like receptor,and nucleotide binding oligomerization domain-like receptor signaling pathway].Taking an intersection of active component targets and silicosis genes,we obtained 61 common genes that were mainly related to the inflammatory response and apoptosis,such as the phosphatidylinositol-3-kinase/protein kinase B signaling pathway,mitogen activated protein kinases signaling pathway,TNF signaling pathway,toll-like receptor signaling pathway,biosynthesis of nitric oxide,and apoptotic process.In the herb-component-gene-pathway network,paeoniflorin,rutin and nobiletin targeted the most genes.In vitro,paeoniflorin,rutin and nobiletin decreased the mRNA levels of inflammatory factors[interleukin(IL)-6,TNF-α,and IL-1β],suppressed p-AKT and cleaved caspase-3,and increased B cell lymphoma(Bcl)-2 protein expression in silica-induced macrophages in a concentration-dependent manner.CONCLUSION:YCF could significantly relieve the inflammatory response of silicosis via suppression of the AKT/Bcl-2/Caspase-3 pathway.
