OBJECTIVE:To determine the effect of Traditional Chinese Medicine(TCM)Fuzheng Xuanfei Huashi prescription(扶正宣肺化湿方,FZXF)on lipopolysaccharide(LPS)-induced pneumonia in mice and identify the mechanism of FZXF in ...OBJECTIVE:To determine the effect of Traditional Chinese Medicine(TCM)Fuzheng Xuanfei Huashi prescription(扶正宣肺化湿方,FZXF)on lipopolysaccharide(LPS)-induced pneumonia in mice and identify the mechanism of FZXF in the treatment of LPS-induced lung inflammation.METHODS:The pneumonia model was established by intraperitoneal injection of 5 mg/kg LPS in mice.Cytokines were detected by enzyme-linked immuneosorbent assay(ELISA),macrophages in lung tissue were determined by immunofluorescence,and pathwayrelated data were determined by quantitative real-time polymerase chain reaction(qPCR)and Western blot.RESULTS:The liver,thymus,and spleen index values and the levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)obviously increased in LPS-treated mice.FZXF decreased the white blood cell count and reduced the increase in the lung wet weight/dry weight ratio caused by LPS.The hematoxylin-eosin staining result showed that FZXF could maintain the integrity of lung tissue structure,alleviate interstitial oedema and alveolar wall thickening,and reduce inflammatory cell infiltration.Moreover,FZXF markedly reduced the expression of proinflammatory cytokines.FZXF also significantly reduced LPS-induced malondialdehyde production and increased superoxide dismutase level in the lung.By immunofluorescence,we found that FZXF could reduce macrophage infiltration.The mRNA expression levels of cyclooxygenase-2(COX-2),prostaglandin E2(PGE2),toll-like receptor 4(TLR4)and nuclear transcription factorκB(NF-κB)in the lung tissue of mice were decreased by treatment with FZXF.In addition,FZXF inhibited the protein expression of TLR4,p-p65 and COX-2.These results indicated that FZXF could inhibit the inflammatory response of LPS induced cytokine storm in mice through TLR4/NF-κB and COX-2/PGE2 signaling pathway.CONCLUSION:These findings were suggested that FZXF prescription suppresses inflammation in LPSinduced pneumonia in mice via TLR4/NF-κB and COX-2/PGE2 pathway.展开更多
The NOD-like receptor family pyrin domain-containing protein 3(NLRP3)inflammasome is an intracellular protein complex containing a nucleotide-binding oligomerization domain,leucine-rich repeats,and a pyrin domain.It i...The NOD-like receptor family pyrin domain-containing protein 3(NLRP3)inflammasome is an intracellular protein complex containing a nucleotide-binding oligomerization domain,leucine-rich repeats,and a pyrin domain.It is a key regulator of inflammation in viral pneumonia(VP).Small-molecule inhibitors targeting various NLRP3 binding sites are advancing into early clinical trials,but their therapeutic utility is incompletely established.Xuanfei Baidu Formula(XF),clinically used for VP treatment,attenuates NLRP3 activation by hampering caspase-11 to impede polarization of pro-inflammatory macrophages in a model of lipopolysaccharide(LPS)-induced lung injury inmice.Herein,we demonstrate that XF attenuated influenza A virus(IAV)-induced lung inflammation as well as lung injury in immunocompetent(but not in macrophage-depleted)mice.RNA sequencing of sorted lung macrophages from IAV-infected mice revealed that XF inhibited activation of the NLRP3 inflammation and interleukin(IL)-1βproduction.Quantitative nuclear magnetic resonance of XF enabled us to develop XF-Comb1,a fixed-ratio combination of five bioactive compounds that recapitulated the bioactivity of XF in suppressing NLRP3 activation in macrophages in vitro and in vivo.Interestingly,XF-Comb1 inhibited assembly of the NLRP3 inflammasome through multi-site interactions with functional residues of NLRP3,apoptosis-associated speck-like protein containing caspase recruitment domain(ASC),and caspase-1.Taken together,this work advances the development of NLRP3 inhibitors by translating a complex herbal formula into defined bioactive compounds.展开更多
Objective:To evaluate the efficacy and safety of XuanFei TongFu method in the treatment of sepsis.Methods:The relevant literatures on the treatment of sepsis by Xuanfei Tongfu method,published by PubMed,Medline and th...Objective:To evaluate the efficacy and safety of XuanFei TongFu method in the treatment of sepsis.Methods:The relevant literatures on the treatment of sepsis by Xuanfei Tongfu method,published by PubMed,Medline and the Cochrane library,CNKI,WEIPU Database,WANFANG Database until December 2019 were searched by computer.Revman 5.3 was used to analyze the relevant literatures that met the inclusion and exclusion criteria,and to evaluate the influence of Xuanfei Tongfu method on gastrointestinal function,adverse reactions of gastrointestinal,28 day mortality.Results:a total of 17 randomized controlled clinical trials(RCTS)involving a total of 984 patients were included.Xuanfei Tongfu decoction combined with the control group can improve intestinal function.Xuanfei Tongfu decoction can reduce gastrointestinal adverse reactions,but there is no statistical significance,and reducing mortality of 28 days(RR=0.64,95%CI(0.42,0.99),P=0.04),reducing the APACHEⅡ[SMD=-0.90,95%CI(-1.49,-0.31),P=0.0003],reducing of Il-6[SMD=-0.36,95%CI(-0.62,-0.1),P=0.007],and improving IL-10 were all better than the control group,the difference was statistically significant.Conclusion:The method of Xuanfei Tongfu can enhance the gastrointestinal function,reduce the gastrointestinal adverse reactions,and improve the prognosis of sepsis patients.It may be achieved by regulating the body's immune inflammatory response.展开更多
Objective:To investigate the molecular mechanism of antiviral effect of Xuanfei Baidu Formula in the treatment of coronavirus disease 2019(COVID-19),and to provide reference for the twreatment of COVID-19 with traditi...Objective:To investigate the molecular mechanism of antiviral effect of Xuanfei Baidu Formula in the treatment of coronavirus disease 2019(COVID-19),and to provide reference for the twreatment of COVID-19 with traditional Chinese medicine.Methods:Traditional Chinese Medicine Systems Pharmacology(TCMSP)was employed to search the effective active component and targets of Xuanfei Baidu Formula.Databases,GeneCards etc.were employed to obtain the relevant target genes of COVID-19 and intersect with the targets of Xuanfei Baidu Formula to obtain the therapeutic targets.The therapeutic targets were uploaded to the STRING database to obtain protein interaction(PPI)information,and Cytoscape was utilized to construct drug-target-disease networks,high-confidence PPI networks for target proteins,and active-target-pathway networks.GO,gene ontology,functional analysis and KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis of potential targets of action were performed using the R package in Bioconductor.The main active components were molecular docking technology with 3C-like protease(3CLPro)and angiotensin-cinverting enzyme2(ACE2)and key targets of SARS-CoV-2,respectively.Results:A total of 99 effective active ingredients of Xuanfei Baidu Formula were selected,of which 22 active ingredients acting on COVID-19 could act on COVID-19 through 52 potential targets.Enrichment yielded 1,364 biological process entries and 22 KEGG pathways in GO,involving paths such as IL-17,JAK-STAT,MAPK,NF-κB,PI3K-Akt,Th1 and Th2 cell differentiation,Th17 cell differentiation,T cell receptor,vascular endothelial growth factor(VEGF)and Salmonella infection.Molecular docking technology results revealed that active components such as luteolin,beta-sitostero,formononetin,and shinpterocarpin in Xuanfei Baidu Formula embraced satisfactory binding to 3CLPro and ACE2,and also had good binding to core targets.Conclusion:Xuanfei Baidu Formula inhibits viral invasion and viral replication mainly by binding to ACE2 and 3CLPro receptors of SARS-CoV-2 through flavonoids and phytosterols,and may play a role in the treatment of COVID-19 by regulating key targets such as IL6,MAPK3,MAPK1,IL1β,CCL2,EGFR,and NOS2 after virus infection of cells,exerting anti-cytokine storm,anti-oxidation,and regulating the body's immunity.展开更多
Objective:This study aims to investigate the therapeutic effects and underlying mechanisms of Xuanfei Baidu Decoction(XFBD)in a mouse model of dampness-heat toxin pneumonia.By exploring how XFBD exerts its effects,we ...Objective:This study aims to investigate the therapeutic effects and underlying mechanisms of Xuanfei Baidu Decoction(XFBD)in a mouse model of dampness-heat toxin pneumonia.By exploring how XFBD exerts its effects,we seek to deepen our understanding of its role in treating pulmonary diseases and to address the current knowledge gap regarding its mechanisms of action,thereby supporting its clinical application.Methods:Ultra-high-performance liquid chromatography and high-resolution mass spectrometry(HRMS)were employed to analyze the chemical constituents of XFBD.The protective effects of XFBD were evaluated using a dampness-heat toxin-induced mouse model,established through dampness-heat exposure and HCoV-229E infection.XFBD was administered orally,followed by assessments including lung index measurement,micro-CT imaging,viral load quantification,cytokine analysis,and histological evaluation via hematoxylin-eosin staining.Proteomics and single-cell transcriptomic analyses were conducted to explore the potential mechanisms underlying XFBD’s pharmacological effects.A cellular model of HCoV-229E infection was developed to investigate changes in the cAMP/PKA signaling pathway.Molecular docking and surface plasmon resonance(SPR)experiments confirmed the strong binding affinity between key XFBD components and PKA.Finally,PKA activators and inhibitors were applied in vitro to validate these mechanistic findings.Results:In vivo studies demonstrated that XFBD significantly reduced the lung index,improved the structural integrity of lung and tongue tissues,and decreased levels of proinflammatory mediators,including IL-6,IL-8,and TNF-α.Proteomic and single-cell transcriptomic analyses showed that the differentially expressed proteins after XFBD treatment were primarily associated with inflammatory responses and immune regulation.The cAMP/PKA signaling pathway was identified as a key mechanism underlying these therapeutic effects.Notably,Western blot,ELISA,molecular docking,and SPR analyses confirmed that XFBD elevated cAMP levels and p-PKA expression,thereby activating the cAMP/PKA signaling pathway in vitro.Conclusion:This study demonstrated that XFBD significantly alleviates symptoms in mice with dampness-heat toxin pneumonia.Its therapeutic effects are mediated,at least in part,through activation of the cAMP/PKA signaling pathway.These findings provide com-pelling evidence that XFBD is an effective herbal remedy against HCoV-229E infection.展开更多
基金Emergency Corona Virus Disease 2019(COVID-19)Response Project of Dongguan:Clinical Efficacy Observation and Mechanism Study of Fuzheng Xuanfei Huashi Formula in the Treatment of COVID-19 Based on the Lingnan Theory of Epidemic Diseases(No.202071715002124)National Natural Science Foundation of China:Study on the Mechanism of Lung Inflammatory Injury Induced by Gut-derived Lipopolysaccharide and Skatole in Spleen Deficiency Animals based on Pulmonary Alveolus Macrophage Heterogeneity(No.82274381)Guangdong Basic and Applied Basic Research Foundation:Development and Industrialization of Traditional Chinese Medicine Classic and Famous Prescription Compound Formulations(No.2021ZD006)。
文摘OBJECTIVE:To determine the effect of Traditional Chinese Medicine(TCM)Fuzheng Xuanfei Huashi prescription(扶正宣肺化湿方,FZXF)on lipopolysaccharide(LPS)-induced pneumonia in mice and identify the mechanism of FZXF in the treatment of LPS-induced lung inflammation.METHODS:The pneumonia model was established by intraperitoneal injection of 5 mg/kg LPS in mice.Cytokines were detected by enzyme-linked immuneosorbent assay(ELISA),macrophages in lung tissue were determined by immunofluorescence,and pathwayrelated data were determined by quantitative real-time polymerase chain reaction(qPCR)and Western blot.RESULTS:The liver,thymus,and spleen index values and the levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)obviously increased in LPS-treated mice.FZXF decreased the white blood cell count and reduced the increase in the lung wet weight/dry weight ratio caused by LPS.The hematoxylin-eosin staining result showed that FZXF could maintain the integrity of lung tissue structure,alleviate interstitial oedema and alveolar wall thickening,and reduce inflammatory cell infiltration.Moreover,FZXF markedly reduced the expression of proinflammatory cytokines.FZXF also significantly reduced LPS-induced malondialdehyde production and increased superoxide dismutase level in the lung.By immunofluorescence,we found that FZXF could reduce macrophage infiltration.The mRNA expression levels of cyclooxygenase-2(COX-2),prostaglandin E2(PGE2),toll-like receptor 4(TLR4)and nuclear transcription factorκB(NF-κB)in the lung tissue of mice were decreased by treatment with FZXF.In addition,FZXF inhibited the protein expression of TLR4,p-p65 and COX-2.These results indicated that FZXF could inhibit the inflammatory response of LPS induced cytokine storm in mice through TLR4/NF-κB and COX-2/PGE2 signaling pathway.CONCLUSION:These findings were suggested that FZXF prescription suppresses inflammation in LPSinduced pneumonia in mice via TLR4/NF-κB and COX-2/PGE2 pathway.
基金supported by the National Natural Science Foundation of China(82141201,82405164,82204878,and 32170872)the Haihe Laboratory of Modern Chinese Medicine(Research and development of a universal treatment formula for respiratory viral infections)+3 种基金the National Key Research and Development Program of China(2021YFC1712905,2021YFC1712904,2020YFA0708004,2021YFE0200300,and 2023YFC2306202)the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202002,ZYYCXTD-D-202001)the China Postdoctoral Science Foundation(2023M742626)the Postdoctoral Fellowship Program of China Postdoctoral Science Foundation(CPSF)(GZC20231927).
文摘The NOD-like receptor family pyrin domain-containing protein 3(NLRP3)inflammasome is an intracellular protein complex containing a nucleotide-binding oligomerization domain,leucine-rich repeats,and a pyrin domain.It is a key regulator of inflammation in viral pneumonia(VP).Small-molecule inhibitors targeting various NLRP3 binding sites are advancing into early clinical trials,but their therapeutic utility is incompletely established.Xuanfei Baidu Formula(XF),clinically used for VP treatment,attenuates NLRP3 activation by hampering caspase-11 to impede polarization of pro-inflammatory macrophages in a model of lipopolysaccharide(LPS)-induced lung injury inmice.Herein,we demonstrate that XF attenuated influenza A virus(IAV)-induced lung inflammation as well as lung injury in immunocompetent(but not in macrophage-depleted)mice.RNA sequencing of sorted lung macrophages from IAV-infected mice revealed that XF inhibited activation of the NLRP3 inflammation and interleukin(IL)-1βproduction.Quantitative nuclear magnetic resonance of XF enabled us to develop XF-Comb1,a fixed-ratio combination of five bioactive compounds that recapitulated the bioactivity of XF in suppressing NLRP3 activation in macrophages in vitro and in vivo.Interestingly,XF-Comb1 inhibited assembly of the NLRP3 inflammasome through multi-site interactions with functional residues of NLRP3,apoptosis-associated speck-like protein containing caspase recruitment domain(ASC),and caspase-1.Taken together,this work advances the development of NLRP3 inhibitors by translating a complex herbal formula into defined bioactive compounds.
基金Key R&D Plan of Shaanxi Provincial Department of Science and Technology(No.2017ZDXM-SF-109)。
文摘Objective:To evaluate the efficacy and safety of XuanFei TongFu method in the treatment of sepsis.Methods:The relevant literatures on the treatment of sepsis by Xuanfei Tongfu method,published by PubMed,Medline and the Cochrane library,CNKI,WEIPU Database,WANFANG Database until December 2019 were searched by computer.Revman 5.3 was used to analyze the relevant literatures that met the inclusion and exclusion criteria,and to evaluate the influence of Xuanfei Tongfu method on gastrointestinal function,adverse reactions of gastrointestinal,28 day mortality.Results:a total of 17 randomized controlled clinical trials(RCTS)involving a total of 984 patients were included.Xuanfei Tongfu decoction combined with the control group can improve intestinal function.Xuanfei Tongfu decoction can reduce gastrointestinal adverse reactions,but there is no statistical significance,and reducing mortality of 28 days(RR=0.64,95%CI(0.42,0.99),P=0.04),reducing the APACHEⅡ[SMD=-0.90,95%CI(-1.49,-0.31),P=0.0003],reducing of Il-6[SMD=-0.36,95%CI(-0.62,-0.1),P=0.007],and improving IL-10 were all better than the control group,the difference was statistically significant.Conclusion:The method of Xuanfei Tongfu can enhance the gastrointestinal function,reduce the gastrointestinal adverse reactions,and improve the prognosis of sepsis patients.It may be achieved by regulating the body's immune inflammatory response.
基金National Natural Science Foundation of China(No.81473592)Changchun University of Traditional Chinese Medicine graduate student“JuJing Cup”academic research and innovation project(No.FK201922)
文摘Objective:To investigate the molecular mechanism of antiviral effect of Xuanfei Baidu Formula in the treatment of coronavirus disease 2019(COVID-19),and to provide reference for the twreatment of COVID-19 with traditional Chinese medicine.Methods:Traditional Chinese Medicine Systems Pharmacology(TCMSP)was employed to search the effective active component and targets of Xuanfei Baidu Formula.Databases,GeneCards etc.were employed to obtain the relevant target genes of COVID-19 and intersect with the targets of Xuanfei Baidu Formula to obtain the therapeutic targets.The therapeutic targets were uploaded to the STRING database to obtain protein interaction(PPI)information,and Cytoscape was utilized to construct drug-target-disease networks,high-confidence PPI networks for target proteins,and active-target-pathway networks.GO,gene ontology,functional analysis and KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis of potential targets of action were performed using the R package in Bioconductor.The main active components were molecular docking technology with 3C-like protease(3CLPro)and angiotensin-cinverting enzyme2(ACE2)and key targets of SARS-CoV-2,respectively.Results:A total of 99 effective active ingredients of Xuanfei Baidu Formula were selected,of which 22 active ingredients acting on COVID-19 could act on COVID-19 through 52 potential targets.Enrichment yielded 1,364 biological process entries and 22 KEGG pathways in GO,involving paths such as IL-17,JAK-STAT,MAPK,NF-κB,PI3K-Akt,Th1 and Th2 cell differentiation,Th17 cell differentiation,T cell receptor,vascular endothelial growth factor(VEGF)and Salmonella infection.Molecular docking technology results revealed that active components such as luteolin,beta-sitostero,formononetin,and shinpterocarpin in Xuanfei Baidu Formula embraced satisfactory binding to 3CLPro and ACE2,and also had good binding to core targets.Conclusion:Xuanfei Baidu Formula inhibits viral invasion and viral replication mainly by binding to ACE2 and 3CLPro receptors of SARS-CoV-2 through flavonoids and phytosterols,and may play a role in the treatment of COVID-19 by regulating key targets such as IL6,MAPK3,MAPK1,IL1β,CCL2,EGFR,and NOS2 after virus infection of cells,exerting anti-cytokine storm,anti-oxidation,and regulating the body's immunity.
基金funded by the National Key R&D Program of China(No.2021YFC1712903)the Science and Technology Innovation Project of the China Academy of ChineseMedical Sciences(No.C12021A04606)the National Natural Science Foundation of China(No.82141206,82151210).
文摘Objective:This study aims to investigate the therapeutic effects and underlying mechanisms of Xuanfei Baidu Decoction(XFBD)in a mouse model of dampness-heat toxin pneumonia.By exploring how XFBD exerts its effects,we seek to deepen our understanding of its role in treating pulmonary diseases and to address the current knowledge gap regarding its mechanisms of action,thereby supporting its clinical application.Methods:Ultra-high-performance liquid chromatography and high-resolution mass spectrometry(HRMS)were employed to analyze the chemical constituents of XFBD.The protective effects of XFBD were evaluated using a dampness-heat toxin-induced mouse model,established through dampness-heat exposure and HCoV-229E infection.XFBD was administered orally,followed by assessments including lung index measurement,micro-CT imaging,viral load quantification,cytokine analysis,and histological evaluation via hematoxylin-eosin staining.Proteomics and single-cell transcriptomic analyses were conducted to explore the potential mechanisms underlying XFBD’s pharmacological effects.A cellular model of HCoV-229E infection was developed to investigate changes in the cAMP/PKA signaling pathway.Molecular docking and surface plasmon resonance(SPR)experiments confirmed the strong binding affinity between key XFBD components and PKA.Finally,PKA activators and inhibitors were applied in vitro to validate these mechanistic findings.Results:In vivo studies demonstrated that XFBD significantly reduced the lung index,improved the structural integrity of lung and tongue tissues,and decreased levels of proinflammatory mediators,including IL-6,IL-8,and TNF-α.Proteomic and single-cell transcriptomic analyses showed that the differentially expressed proteins after XFBD treatment were primarily associated with inflammatory responses and immune regulation.The cAMP/PKA signaling pathway was identified as a key mechanism underlying these therapeutic effects.Notably,Western blot,ELISA,molecular docking,and SPR analyses confirmed that XFBD elevated cAMP levels and p-PKA expression,thereby activating the cAMP/PKA signaling pathway in vitro.Conclusion:This study demonstrated that XFBD significantly alleviates symptoms in mice with dampness-heat toxin pneumonia.Its therapeutic effects are mediated,at least in part,through activation of the cAMP/PKA signaling pathway.These findings provide com-pelling evidence that XFBD is an effective herbal remedy against HCoV-229E infection.
