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 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
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.
