Background:Hua-Yi-Jie-Du formula(HYJD)is a traditional Chinese medicine that has proven effective against viral pneumonia and was extensively used during the COVID-19 pandemic.This study investigates how HYJD influenc...Background:Hua-Yi-Jie-Du formula(HYJD)is a traditional Chinese medicine that has proven effective against viral pneumonia and was extensively used during the COVID-19 pandemic.This study investigates how HYJD influences group 2 innate lymphoid cell(ILC2)and nucleotide oligomerization domain(NOD)-like receptor protein 3(NLRP3)inflammasome activation in a mouse model of viral pneumonia.Methods:A mouse model of viral pneumonia was established through the administration of polyinosinic-polycytidylic acid(poly(I:C))via nasal drops.Histopathological analysis of lung tissue was conducted,alongside enzyme-linked immunosorbent assay to quantify cytokine levels in serum and bronchoalveolar lavage fluid(BALF).Flow cytometry was employed to detect ILC2 cells in lung tissue and spleen,while immunofluorescence techniques were utilized to visualize ILC2 cells in lung tissue.Transcriptomic sequencing was performed,and the results were validated using qRT-PCR and western blot analysis.Results:HYJD significantly ameliorated inflammatory infiltration in lung tissue,decreased mucus protein secretion,and reduced the serum levels of inflammatory cytokines interleukin(IL)-1β,IL-6,and tumor necrosis factor-alpha(TNF-α).Additionally,it lowered the expression of cytokines IL-4,IL-5,IL-13,IL-25,thymic stromal lymphopoietin(TSLP),and IL-33 in BALF,and reduced the differentiation of ILC2 cells in both lung tissue and spleen.Transcriptomic analysis and experimental validation revealed that HYJD downregulated the expression of NLRP3 related genes and proteins within the NOD-like receptor signaling pathway.Conclusion:The mechanism by which HYJD intervenes in acute lung injury associated with viral pneumonia may involve the reduction of ILC2 cells differentiation and the inhibition of NLRP3 activation.展开更多
The role of NF-κB and the NLRP3 inflammasome in the chronic inflammatory microenvironment of non-alcoholic steatohepatitis(NASH)has been posited as crucial.The Yanggan Jiangmei Formula(YGJMF)has shown promise in amel...The role of NF-κB and the NLRP3 inflammasome in the chronic inflammatory microenvironment of non-alcoholic steatohepatitis(NASH)has been posited as crucial.The Yanggan Jiangmei Formula(YGJMF)has shown promise in ameliorating hepatic steatosis in NASH patients,yet its pharmacological mechanisms remain largely unexplored.This study was conducted to investigate the efficacy of YGJMF in NASH and to elucidate its pharmacological underpinnings.To simulate NASH both in vivo and in vitro,high-fat-diet(HFD)rats and HepG2 cells stimulated with free fatty acids(FFAs)were utilized.The severity of liver injury and lipid deposition was assessed using serum indicators,histopathological staining,micro-magnetic resonance imaging(MRI),and the liver-tomuscle signal intensity ratio(SIRL/M).Furthermore,a combination of enzyme-linked immunosorbent assay(ELISA),immunohistochemistry(IHC),immunofluorescence,real-time quantitative polymerase chain reaction(RT-qPCR),and Western blotting analyses was employed to investigate the NF-κB/NLRP3 signaling pathway and associated cytokine levels.The results from liver pathology,MRI assessments,and biochemical tests in rat models demonstrated YGJMF’s significant effectiveness in reducing liver damage and lipid accumulation.Additionally,YGJMF markedly reduced hepatocyte inflammation by downregulating inflammatory cytokines in both liver tissue and serum.Furthermore,YGJMF was found to disrupt NF-κB activation,consequently inhibiting the assembly of the NLRP3 inflammasome in both the in vitro and in vivo models.The preliminary findings of this study suggest that YGJMF may alleviate hepatic steatosis and inhibit the NF-κB/NLRP3 signaling pathway,thereby exerting anti-inflammatory effects in NASH.展开更多
Objective:To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula(HTQS for-mula),for the health management and treatment of non-alcoholic fatty liver disease(NAFLD).Methods:A rat model of NAFLD was ...Objective:To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula(HTQS for-mula),for the health management and treatment of non-alcoholic fatty liver disease(NAFLD).Methods:A rat model of NAFLD was employed to examine the efficacy and safety of the HTQS formula.In vivo active components and potential mechanisms of the HTQS formula were identified using UPLC‒MS/MS combined with network pharmacology.The influence of the HTQS formula on the dominating proteins in PI3K/Akt pathway was validated in vivo using western blot.Finally,16S rRNA sequencing of the gut microbiome was conducted followed by targeted metabolomics detecting fecal short-chain fatty acids(SCFAs)and bile acids to determine the impact of the HTQS formula on gut microbiota.Results:The HTQS formula reduced weight gain and hepatic steatosis in NAFLD rats and decreased serum total cholesterol(TC),triglycerides,blood glucose,and insulin resistance(IR)without causing liver or kidney injury.We detected 28 components using UPLC‒MS/MS and identified 439 shared targets be-tween NAFLD and the HTQS formula.Primarily,we focused on the PI3K/Akt signaling pathway based on protein‒protein interaction network analysis.We validated that the HTQS formula inhibited liver stea-tosis and inflammation by increasing the phosphorylation levels of PI3K,AKT,P27,GSK3b in the PI3K/Akt signaling pathway.16S rRNA sequencing revealed that the HTQS formula reduced the abundance of the genus Family_XIII_AD3011_group,which was positively correlated with IR and taurodeoxycholic acid.In addition,Lachnospiraceae_UCG_010 inversely correlated with TC and five bile acids,which could be essential to the therapeutic effect of the HTQS formula against NAFLD.Conclusions:The HTQS formula proved to be an effective pharmacotherapy for NAFLD without causing liver or kidney injury.Multiple potent components of the HTQS formula could alleviate liver steatosis and lipid metabolism disorder by modulating the PI3K/Akt signaling pathway and restoring gut microbiota composition.展开更多
This paper reviews the related formulas of Sugemule,introduces the advances in research of clinical application of these formulas in treating Heyi type insomnia,cardiac diseases,and renal diseases.Besides,it summarize...This paper reviews the related formulas of Sugemule,introduces the advances in research of clinical application of these formulas in treating Heyi type insomnia,cardiac diseases,and renal diseases.Besides,it summarizes pharmacological research advances regarding these formulas,including their anti-insomnia effects,cardioprotective properties,and ovarian function preservation capabilities.This study is expected to provide a reference and insight for in-depth and systematic research on classical Mongolian medicinal formulas.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
Background:This investigation aimed to evaluate the therapeutic impact of the Yigan Xiaozheng formula on liver cirrhosis in rats,particularly induced by diethylnitrosamine(DEN).The study focused on analyzing liver str...Background:This investigation aimed to evaluate the therapeutic impact of the Yigan Xiaozheng formula on liver cirrhosis in rats,particularly induced by diethylnitrosamine(DEN).The study focused on analyzing liver structure,cell apoptosis,and the modulation of the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway,employing a combination of network pharmacology and experimental approaches.Methods:A DEN-induced rat model of liver cirrhosis was established to assess the formula’s effectiveness.Parameters such as overall health,liver morphology,and survival were monitored.Network pharmacology was employed to decipher the active compounds and key targets of the formula in addressing liver cirrhosis.Predictions made via network pharmacology were substantiated through experimental validation in the animal model.Results:Administration of the Yigan Xiaozheng formula led to noticeable improvements in clinical symptoms of liver cirrhosis in rats,marked by enhanced body weight,lessened liver pathology,and higher survival rates.Network pharmacological analysis unveiled intricate interactions between active ingredients of the formula and cirrhosis-related targets.Protein-protein interaction(PPI)networks pinpointed crucial proteins and regulatory modules.Enrichment analysis underscored a significant involvement of the JAK2/STAT3 signaling pathway.On a molecular scale,the formula was observed to reduce the expression of BCL-2 associated X protein(Bax)and cytochrome C(Cyt-C),diminish the Bax/B-cell lymphoma 2(Bcl-2)ratio,and impede JAK2/STAT3 pathway activation,thereby curtailing liver fibrosis and cellular apoptosis.Conclusion:The study demonstrates the Yigan Xiaozheng formula’s capacity to ameliorate liver cirrhosis in a DEN-induced model,primarily through its active ingredients’interactions with cirrhosis targets and modulation of the JAK2/STAT3 pathway.These findings endorse the potential of this traditional Chinese medicinal formula as a viable treatment option for liver cirrhosis.展开更多
Objective To investigate the effects of Zuogui Jiangtang Yishen Formula(左归降糖益肾方,ZGJTYSF)in regulating the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1/gasdermin D(GSDMD)sig...Objective To investigate the effects of Zuogui Jiangtang Yishen Formula(左归降糖益肾方,ZGJTYSF)in regulating the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1/gasdermin D(GSDMD)signaling axis on pyroptosis in rats with diabetic kidney disease(DKD).Methods Fifty male specific pathogen-free(SPF)grade Goto-Kakizaki(GK)rats(12 weeks old)were fed a high-fat diet for one month to establish an early DKD model.Model establishment was confirmed when fasting blood glucose(FBG)≥11.1 mmol/L and urinary albuminto-creatinine ratio(uACR)≥30 mg/g.The successfully modeled early DKD rats were randomly divided by random number table into five groups(n=10 per group):model group;dapagliflozin group(1.0 mg/kg,by gavage,served as positive control);and low-,medium-,and high-dose of ZGJTYSF groups(4.9,9.9,and 19.9 g/kg,respectively,by gavage).Age-matched male SPF Wistar rats(n=10)served as control group.Rats in control and model groups were gavaged with equivalent volumes of distilled water.Treatment lasted 12 weeks.Changes in uACR,FBG,and renal function were observed in all groups.Hematoxylin-eosin(HE),periodic acid-Schiff(PAS),and Masson staining were used to observe renal histopathological changes.Immunohistochemistry was performed to detect the localization and expression of caspase-1,GSDMD,and NLRP3 in rat renal tissues.Terminal deoxynucleotidyl transferase deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL)was utilized to detect pyroptosis in renal tissues.Quantitative real-time polymerase chain reaction(qPCR)and Western blot were applied to detect mRNA and protein expression levels of NLRP3,caspase-1,GSDMD,interleukin(IL)-1β,and IL-18.Results Compared with model group,all doses of ZGJTYSF showed reductions in FBG,with medium-and high-dose of ZGJTYSF groups demonstrating significant decreases at week 8 and 12(P<0.05).For uACR,all doses of ZGJTYSF groups exhibited a decreasing trend,with high-dose of ZGJTYSF group being significantly lower than low-and medium-dose of ZGJTYSF groups at week 12(P<0.05)and showing no significant difference from dapagliflozin group(P>0.05).No significant differences in renal function parameters(serum creatinine,blood urea nitrogen,and uric acid)were observed among groups(P>0.05).Histopathological examination revealed milder glomerular and tubular lesions in both ZGJTYSF groups and dapagliflozin group,with renal pathological changes in high-dose of ZGJTYSF group resembling those in dapagliflozin group.Immunohistochemistry demonstrated significantly reduced expression of caspase-1,GSDMD,and NLRP3 in renal tissues of dapagliflozin group and high-dose of ZGJTYSF group compared with model group(P<0.05 or P<0.01),while the differences in low-and medium-dose of ZGJTYSF groups were not statistically significant(P>0.05).TUNEL assay showed significantly fewer TUNEL-positive cells in renal tissues of dapagliflozin and high-dose of ZGJTYSF groups(P<0.01),indicating a marked reduction in pyroptotic cells.Molecular analysis revealed that compared with model group,both dapagliflozin and high-dose of ZGJTYSF groups showed significantly downregulated mRNA and protein expression levels of NLRP3,caspase-1,GSDMD,IL-1β,and IL-18 in renal tissues(P<0.01),while low-and medium-dose of ZGJTYSF groups showed downward trends without statistical significance(P>0.05).Conclusion ZGJTYSF may inhibit renal pyroptosis by regulating the NLRP3/caspase-1/GSDMD signaling axis,thereby preventing and treating early renal injury in DKD and delaying the onset and progression of DKD.展开更多
OBJECTIVE:To explore the bioactive constituents,key targets,signalling pathways,and molecular mechanisms of Lianshi Jianpi formula(莲实健脾方,LSJPF)in the treatment of impaired glucose tolerance(IGT)through network ph...OBJECTIVE:To explore the bioactive constituents,key targets,signalling pathways,and molecular mechanisms of Lianshi Jianpi formula(莲实健脾方,LSJPF)in the treatment of impaired glucose tolerance(IGT)through network pharmacology,molecular docking,and in vivo experiments.METHODS:The active ingredients and targets of LSJPF were identified using the Traditional Chinese Medicine Systems Pharmacology and HERB databases,whereas the IGT-related targets were sourced from Gene Cards,Dis Ge NET,and Pub Med.The overlap analysis identified potential targets of LSJPF.Protein-protein interaction networks and core targets were evaluated using the Search Tool for the Retrieval of Interacting Genes/Proteins and Cytoscape,and molecular docking confirmed the binding affinities.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed using Metascape.The therapeutic mechanisms were validated in an animal IGT model.RESULTS:LSJPF contained 229 compounds,with 15 active compounds and 77 potential target proteins.The phosphatidylinositol-3-kinase(PI3K)-protein kinase B(AKT)signalling pathway emerged as a key IGT pathway.The KEGG enrichment analysis revealed the pivotal genes RAC-alpha serine/threonine-protein kinase(AKT1),heat shock protein 90 k Da alpha B1,and B-cell lymphoma 2 family protein,which predominantly interact with beta-sitosterol and beta-carotene,the major constituents of Semen Euryales,Semen lablab Album,Semen sojae Atricolor in LSJPF.Molecular docking revealed strong binding affinities between LSJPF and IGT-related targets.In an animal IGT model,LSJPF treatment prevented weight loss;reduced food and water intake;decreased blood glucose levels;improved insulin resistance;decreased serum triglyceride,cholesterol,and low-density lipoprotein cholesterol levels;alleviated liver pathology;and significantly increased the levels of phosphorylated adenosine 5'-monophosphate-activated protein kinase(AMPK),PI3K,and AKT,suggesting its potential role in regulating glucose and lipid metabolism.CONCLUSIONS:These findings reveal the potential of LSJPF as an IGT intervention that targets the AMPK/PI3K/AKT cascade,validating network pharmacology predictions and highlighting the role of multipathway mechanisms in metabolic diseases.展开更多
基金supported by Yunnan Provincial Major Science and Technology Special Program(No.202402AA310035)Yunnan Key Laboratory of Dai and Yi Medicines(Yunnan University of Chinese Medicine)(No.2024JS2404)Research Foundation of Chuxiong Medical College(No.2024YYXM38).
文摘Background:Hua-Yi-Jie-Du formula(HYJD)is a traditional Chinese medicine that has proven effective against viral pneumonia and was extensively used during the COVID-19 pandemic.This study investigates how HYJD influences group 2 innate lymphoid cell(ILC2)and nucleotide oligomerization domain(NOD)-like receptor protein 3(NLRP3)inflammasome activation in a mouse model of viral pneumonia.Methods:A mouse model of viral pneumonia was established through the administration of polyinosinic-polycytidylic acid(poly(I:C))via nasal drops.Histopathological analysis of lung tissue was conducted,alongside enzyme-linked immunosorbent assay to quantify cytokine levels in serum and bronchoalveolar lavage fluid(BALF).Flow cytometry was employed to detect ILC2 cells in lung tissue and spleen,while immunofluorescence techniques were utilized to visualize ILC2 cells in lung tissue.Transcriptomic sequencing was performed,and the results were validated using qRT-PCR and western blot analysis.Results:HYJD significantly ameliorated inflammatory infiltration in lung tissue,decreased mucus protein secretion,and reduced the serum levels of inflammatory cytokines interleukin(IL)-1β,IL-6,and tumor necrosis factor-alpha(TNF-α).Additionally,it lowered the expression of cytokines IL-4,IL-5,IL-13,IL-25,thymic stromal lymphopoietin(TSLP),and IL-33 in BALF,and reduced the differentiation of ILC2 cells in both lung tissue and spleen.Transcriptomic analysis and experimental validation revealed that HYJD downregulated the expression of NLRP3 related genes and proteins within the NOD-like receptor signaling pathway.Conclusion:The mechanism by which HYJD intervenes in acute lung injury associated with viral pneumonia may involve the reduction of ILC2 cells differentiation and the inhibition of NLRP3 activation.
基金supported by the National Natural Science Foundation of China(No.8200151236)the Postgraduate Research Practice Innovation Program of Jiangsu Province(No.KYCX22_1903)the Outstanding Clinical Talents of Traditional Chinese Medicine in Jiangsu Province(No.18,Jiangsu Science and Education of Traditional Chinese Medicine[2017]).
文摘The role of NF-κB and the NLRP3 inflammasome in the chronic inflammatory microenvironment of non-alcoholic steatohepatitis(NASH)has been posited as crucial.The Yanggan Jiangmei Formula(YGJMF)has shown promise in ameliorating hepatic steatosis in NASH patients,yet its pharmacological mechanisms remain largely unexplored.This study was conducted to investigate the efficacy of YGJMF in NASH and to elucidate its pharmacological underpinnings.To simulate NASH both in vivo and in vitro,high-fat-diet(HFD)rats and HepG2 cells stimulated with free fatty acids(FFAs)were utilized.The severity of liver injury and lipid deposition was assessed using serum indicators,histopathological staining,micro-magnetic resonance imaging(MRI),and the liver-tomuscle signal intensity ratio(SIRL/M).Furthermore,a combination of enzyme-linked immunosorbent assay(ELISA),immunohistochemistry(IHC),immunofluorescence,real-time quantitative polymerase chain reaction(RT-qPCR),and Western blotting analyses was employed to investigate the NF-κB/NLRP3 signaling pathway and associated cytokine levels.The results from liver pathology,MRI assessments,and biochemical tests in rat models demonstrated YGJMF’s significant effectiveness in reducing liver damage and lipid accumulation.Additionally,YGJMF markedly reduced hepatocyte inflammation by downregulating inflammatory cytokines in both liver tissue and serum.Furthermore,YGJMF was found to disrupt NF-κB activation,consequently inhibiting the assembly of the NLRP3 inflammasome in both the in vitro and in vivo models.The preliminary findings of this study suggest that YGJMF may alleviate hepatic steatosis and inhibit the NF-κB/NLRP3 signaling pathway,thereby exerting anti-inflammatory effects in NASH.
基金supported by the General Program of National Natural Science Foundation of China(82374308)National Key Research and Development Program(NKRDP)(2022YFC2010104)Henan Province Special Projects of Traditional Chinese Medicine Science Research(2024ZY2067),and National Talent Precision Cultivation Plan of the Beijing University of Chinese Medicine.
文摘Objective:To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula(HTQS for-mula),for the health management and treatment of non-alcoholic fatty liver disease(NAFLD).Methods:A rat model of NAFLD was employed to examine the efficacy and safety of the HTQS formula.In vivo active components and potential mechanisms of the HTQS formula were identified using UPLC‒MS/MS combined with network pharmacology.The influence of the HTQS formula on the dominating proteins in PI3K/Akt pathway was validated in vivo using western blot.Finally,16S rRNA sequencing of the gut microbiome was conducted followed by targeted metabolomics detecting fecal short-chain fatty acids(SCFAs)and bile acids to determine the impact of the HTQS formula on gut microbiota.Results:The HTQS formula reduced weight gain and hepatic steatosis in NAFLD rats and decreased serum total cholesterol(TC),triglycerides,blood glucose,and insulin resistance(IR)without causing liver or kidney injury.We detected 28 components using UPLC‒MS/MS and identified 439 shared targets be-tween NAFLD and the HTQS formula.Primarily,we focused on the PI3K/Akt signaling pathway based on protein‒protein interaction network analysis.We validated that the HTQS formula inhibited liver stea-tosis and inflammation by increasing the phosphorylation levels of PI3K,AKT,P27,GSK3b in the PI3K/Akt signaling pathway.16S rRNA sequencing revealed that the HTQS formula reduced the abundance of the genus Family_XIII_AD3011_group,which was positively correlated with IR and taurodeoxycholic acid.In addition,Lachnospiraceae_UCG_010 inversely correlated with TC and five bile acids,which could be essential to the therapeutic effect of the HTQS formula against NAFLD.Conclusions:The HTQS formula proved to be an effective pharmacotherapy for NAFLD without causing liver or kidney injury.Multiple potent components of the HTQS formula could alleviate liver steatosis and lipid metabolism disorder by modulating the PI3K/Akt signaling pathway and restoring gut microbiota composition.
基金Supported by Laboratory Open Fund Project of Inner Mongolia Medical University in 2024(2024ZN20)Key Laboratory of Mongolian Medicine Open Fund Project of Inner Mongolia Autonomous Region(GX20240003)+2 种基金Inner Mongolia Medicine Collaborative Innovation Center Achievement Transformation and Cultivation Project(MYYXTPY202402)Project of Construction of Key Laboratory of Mongolian Medicine in Universities of Inner Mongolia Autonomous Region(RZ2400000451)Project of Construction of Key Laboratory of Mongolian Medicine Research Platform in Inner Mongolia Autonomous Region(DC2400000724).
文摘This paper reviews the related formulas of Sugemule,introduces the advances in research of clinical application of these formulas in treating Heyi type insomnia,cardiac diseases,and renal diseases.Besides,it summarizes pharmacological research advances regarding these formulas,including their anti-insomnia effects,cardioprotective properties,and ovarian function preservation capabilities.This study is expected to provide a reference and insight for in-depth and systematic research on classical Mongolian medicinal formulas.
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supported by National Natural Science Foundation of China Grant Program(No.81603555).
文摘Background:This investigation aimed to evaluate the therapeutic impact of the Yigan Xiaozheng formula on liver cirrhosis in rats,particularly induced by diethylnitrosamine(DEN).The study focused on analyzing liver structure,cell apoptosis,and the modulation of the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway,employing a combination of network pharmacology and experimental approaches.Methods:A DEN-induced rat model of liver cirrhosis was established to assess the formula’s effectiveness.Parameters such as overall health,liver morphology,and survival were monitored.Network pharmacology was employed to decipher the active compounds and key targets of the formula in addressing liver cirrhosis.Predictions made via network pharmacology were substantiated through experimental validation in the animal model.Results:Administration of the Yigan Xiaozheng formula led to noticeable improvements in clinical symptoms of liver cirrhosis in rats,marked by enhanced body weight,lessened liver pathology,and higher survival rates.Network pharmacological analysis unveiled intricate interactions between active ingredients of the formula and cirrhosis-related targets.Protein-protein interaction(PPI)networks pinpointed crucial proteins and regulatory modules.Enrichment analysis underscored a significant involvement of the JAK2/STAT3 signaling pathway.On a molecular scale,the formula was observed to reduce the expression of BCL-2 associated X protein(Bax)and cytochrome C(Cyt-C),diminish the Bax/B-cell lymphoma 2(Bcl-2)ratio,and impede JAK2/STAT3 pathway activation,thereby curtailing liver fibrosis and cellular apoptosis.Conclusion:The study demonstrates the Yigan Xiaozheng formula’s capacity to ameliorate liver cirrhosis in a DEN-induced model,primarily through its active ingredients’interactions with cirrhosis targets and modulation of the JAK2/STAT3 pathway.These findings endorse the potential of this traditional Chinese medicinal formula as a viable treatment option for liver cirrhosis.
基金National Natural Science Foundation of China(U21A20411)Innovative Research Group Program of Hunan Provincial Natural Science Foundation (2024JJ1007)Hunan Provincial Natural Science Foundation(2023JJ30473)。
文摘Objective To investigate the effects of Zuogui Jiangtang Yishen Formula(左归降糖益肾方,ZGJTYSF)in regulating the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1/gasdermin D(GSDMD)signaling axis on pyroptosis in rats with diabetic kidney disease(DKD).Methods Fifty male specific pathogen-free(SPF)grade Goto-Kakizaki(GK)rats(12 weeks old)were fed a high-fat diet for one month to establish an early DKD model.Model establishment was confirmed when fasting blood glucose(FBG)≥11.1 mmol/L and urinary albuminto-creatinine ratio(uACR)≥30 mg/g.The successfully modeled early DKD rats were randomly divided by random number table into five groups(n=10 per group):model group;dapagliflozin group(1.0 mg/kg,by gavage,served as positive control);and low-,medium-,and high-dose of ZGJTYSF groups(4.9,9.9,and 19.9 g/kg,respectively,by gavage).Age-matched male SPF Wistar rats(n=10)served as control group.Rats in control and model groups were gavaged with equivalent volumes of distilled water.Treatment lasted 12 weeks.Changes in uACR,FBG,and renal function were observed in all groups.Hematoxylin-eosin(HE),periodic acid-Schiff(PAS),and Masson staining were used to observe renal histopathological changes.Immunohistochemistry was performed to detect the localization and expression of caspase-1,GSDMD,and NLRP3 in rat renal tissues.Terminal deoxynucleotidyl transferase deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL)was utilized to detect pyroptosis in renal tissues.Quantitative real-time polymerase chain reaction(qPCR)and Western blot were applied to detect mRNA and protein expression levels of NLRP3,caspase-1,GSDMD,interleukin(IL)-1β,and IL-18.Results Compared with model group,all doses of ZGJTYSF showed reductions in FBG,with medium-and high-dose of ZGJTYSF groups demonstrating significant decreases at week 8 and 12(P<0.05).For uACR,all doses of ZGJTYSF groups exhibited a decreasing trend,with high-dose of ZGJTYSF group being significantly lower than low-and medium-dose of ZGJTYSF groups at week 12(P<0.05)and showing no significant difference from dapagliflozin group(P>0.05).No significant differences in renal function parameters(serum creatinine,blood urea nitrogen,and uric acid)were observed among groups(P>0.05).Histopathological examination revealed milder glomerular and tubular lesions in both ZGJTYSF groups and dapagliflozin group,with renal pathological changes in high-dose of ZGJTYSF group resembling those in dapagliflozin group.Immunohistochemistry demonstrated significantly reduced expression of caspase-1,GSDMD,and NLRP3 in renal tissues of dapagliflozin group and high-dose of ZGJTYSF group compared with model group(P<0.05 or P<0.01),while the differences in low-and medium-dose of ZGJTYSF groups were not statistically significant(P>0.05).TUNEL assay showed significantly fewer TUNEL-positive cells in renal tissues of dapagliflozin and high-dose of ZGJTYSF groups(P<0.01),indicating a marked reduction in pyroptotic cells.Molecular analysis revealed that compared with model group,both dapagliflozin and high-dose of ZGJTYSF groups showed significantly downregulated mRNA and protein expression levels of NLRP3,caspase-1,GSDMD,IL-1β,and IL-18 in renal tissues(P<0.01),while low-and medium-dose of ZGJTYSF groups showed downward trends without statistical significance(P>0.05).Conclusion ZGJTYSF may inhibit renal pyroptosis by regulating the NLRP3/caspase-1/GSDMD signaling axis,thereby preventing and treating early renal injury in DKD and delaying the onset and progression of DKD.
基金Supported by Natural Science Foundation of Fujian Province Project:To Investigate the Mechanism of Lianshi Jianpi Formula In Improving Impaired Glucose Tolerance via the Fibroblast Growth Factor 21-Mediated Adenosine 5‘-monophosphate-Activated Protein Kinases Energy Metabolism Pathway(No. 2023J01841)ZHOU Guoying Fujian Province Famous Traditional Chinese Medicine Inheritance Workstation Construction Project(Min Zhong Yi[2023] No. 56)。
文摘OBJECTIVE:To explore the bioactive constituents,key targets,signalling pathways,and molecular mechanisms of Lianshi Jianpi formula(莲实健脾方,LSJPF)in the treatment of impaired glucose tolerance(IGT)through network pharmacology,molecular docking,and in vivo experiments.METHODS:The active ingredients and targets of LSJPF were identified using the Traditional Chinese Medicine Systems Pharmacology and HERB databases,whereas the IGT-related targets were sourced from Gene Cards,Dis Ge NET,and Pub Med.The overlap analysis identified potential targets of LSJPF.Protein-protein interaction networks and core targets were evaluated using the Search Tool for the Retrieval of Interacting Genes/Proteins and Cytoscape,and molecular docking confirmed the binding affinities.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed using Metascape.The therapeutic mechanisms were validated in an animal IGT model.RESULTS:LSJPF contained 229 compounds,with 15 active compounds and 77 potential target proteins.The phosphatidylinositol-3-kinase(PI3K)-protein kinase B(AKT)signalling pathway emerged as a key IGT pathway.The KEGG enrichment analysis revealed the pivotal genes RAC-alpha serine/threonine-protein kinase(AKT1),heat shock protein 90 k Da alpha B1,and B-cell lymphoma 2 family protein,which predominantly interact with beta-sitosterol and beta-carotene,the major constituents of Semen Euryales,Semen lablab Album,Semen sojae Atricolor in LSJPF.Molecular docking revealed strong binding affinities between LSJPF and IGT-related targets.In an animal IGT model,LSJPF treatment prevented weight loss;reduced food and water intake;decreased blood glucose levels;improved insulin resistance;decreased serum triglyceride,cholesterol,and low-density lipoprotein cholesterol levels;alleviated liver pathology;and significantly increased the levels of phosphorylated adenosine 5'-monophosphate-activated protein kinase(AMPK),PI3K,and AKT,suggesting its potential role in regulating glucose and lipid metabolism.CONCLUSIONS:These findings reveal the potential of LSJPF as an IGT intervention that targets the AMPK/PI3K/AKT cascade,validating network pharmacology predictions and highlighting the role of multipathway mechanisms in metabolic diseases.
