BACKGROUND Helicobacter pylori(HP),the most common pathogenic microorganism in stomach,can induce inflammatory reactions in the gastric mucosa,causing chronic gastritis and even gastric cancer.HP infection affects ove...BACKGROUND Helicobacter pylori(HP),the most common pathogenic microorganism in stomach,can induce inflammatory reactions in the gastric mucosa,causing chronic gastritis and even gastric cancer.HP infection affects over 4.4 billion people globally,with a worldwide infection rate of up to 50%.The multidrug resistance of HP poses a serious challenge to eradication.It has been monstrated that compared to bismuth quadruple therapy,Qingre Huashi decoction(QHD)combined with triple therapy exhibits comparable eradication rates but with a lower incidence of adverse reactions;in addition,QHD directly inhibit and kill HP in vitro.METHODS In this study,12 HP strains were isolated in vitro after biopsy during gastroscopy of HP-infected patients.In vitro,the minimum inhibitory concentration(MIC)values for clinical HP strains and biofilm quantification were determined through the E-test method and crystal violet staining,respectively.The most robust biofilm-forming strain of HP was selected,and QHD was evaluated for its inhibitory and bactericidal effects on the strain with strong biofilm formation.This assessment was performed using agar dilution,E-test,killing dynamics,and transmission electron microscopy(TEM).The study also explored the impact of QHD on antibiotic resistance in these HP strains with strong biofilm formation.Crystalline violet method,scanning electron microscopy,laser confocal scanning microscopy,and(p)ppGpp chromatographic identification were employed to evaluate the effect of QHD on biofilm in strong biofilm-forming HP strains.The effect of QHD on biofilm and efflux pump-related gene expression was evaluated by quantitative polymerase chain reaction.Non-targeted metabolomics with UHPLC-MS/MS was used to identify potential metabolic pathways and biomarkers which were different between the NC and QHD groups.RESULTS HP could form biofilms of different degrees in vitro,and the intensity of formation was associated with the drug resistance of the strain.QHD had strong bacteriostatic and bactericidal effects on HP,with MICs of 32-64 mg/mL.QHD could inhibit the biofilm formation of the strong biofilm-forming HP strains,disrupt the biofilm structure,lower the accumulation of(p)ppGpp,decrease the expression of biofilm-related genes including LuxS,Spot,glup(HP1174),NapA,and CagE,and reduce the expression of efflux pump-related genes such as HP0605,HP0971,HP1327,and HP1489.Based on metabolomic analysis,QHD induced oxidative stress in HP,enhanced metabolism,and potentially inhibited relevant signaling pathways by upregulating adenosine monophosphate(AMP),thereby affecting HP growth,metabolism,and protein synthesis.CONCLUSION QHD exerts bacteriostatic and bactericidal effects on HP,and reduces HP drug resistance by inhibiting HP biofilm formation,destroying its biofilm structure,inhibiting the expression of biofilm-related genes and efflux pump-related genes,enhancing HP metabolism,and activating AMP in HP.展开更多
Objective:To explore the target and signal pathway of Qingchang Huashi Decoction(QCHSD)in the treatment of ulcerative colitis(UC)by using network pharmacology,so as to explain its molecular mechanism of action in the ...Objective:To explore the target and signal pathway of Qingchang Huashi Decoction(QCHSD)in the treatment of ulcerative colitis(UC)by using network pharmacology,so as to explain its molecular mechanism of action in the treatment of UC from damp heat.Methods:TCMSP was used to screen the potential active components(OB≥30%,DL≥0.18)and target of QCHSD.The network of"potential active ingredients-target-disease"was constructed by using the database of TCMSP and GeneCards.Using the string platform,the protein protein interaction(PPI)network model was constructed to find the core target.Go and KEGG enrichment of potential targets were analyzed by R software.Results:The results of network analysis showed that quercetin,kaempferol,scutellarin and baicalein were the top four active ingredients in QCHSD.210 gene targets were found in QCHSD,4213 in UC.The key targets of QCHSD in treating UC included AKT1,IL-6,VEGFA,CASP3,etc.GO enrichment analysis showed that these gene targets mainly affected nuclear receptor,steroid receptor,cytokine receptor binding,cytokine activity,etc.KEGG enrichment analysis showed that AGE-RAGE signaling pathway,IL-17 signaling pathway and TNF were more abundant.Conclusion:This study describes the material basis and mechanism of QCHSD in the treatment of UC,which provides theoretical basis and research direction for future research.展开更多
基金Supported by the National Natural Science Foundation of China,No.81973615 and No.82304930Natural Science Foundation of Beijing,No.7332323Capital’s Funds for Health Improvement and Research,No.CF2022-2-40711.
文摘BACKGROUND Helicobacter pylori(HP),the most common pathogenic microorganism in stomach,can induce inflammatory reactions in the gastric mucosa,causing chronic gastritis and even gastric cancer.HP infection affects over 4.4 billion people globally,with a worldwide infection rate of up to 50%.The multidrug resistance of HP poses a serious challenge to eradication.It has been monstrated that compared to bismuth quadruple therapy,Qingre Huashi decoction(QHD)combined with triple therapy exhibits comparable eradication rates but with a lower incidence of adverse reactions;in addition,QHD directly inhibit and kill HP in vitro.METHODS In this study,12 HP strains were isolated in vitro after biopsy during gastroscopy of HP-infected patients.In vitro,the minimum inhibitory concentration(MIC)values for clinical HP strains and biofilm quantification were determined through the E-test method and crystal violet staining,respectively.The most robust biofilm-forming strain of HP was selected,and QHD was evaluated for its inhibitory and bactericidal effects on the strain with strong biofilm formation.This assessment was performed using agar dilution,E-test,killing dynamics,and transmission electron microscopy(TEM).The study also explored the impact of QHD on antibiotic resistance in these HP strains with strong biofilm formation.Crystalline violet method,scanning electron microscopy,laser confocal scanning microscopy,and(p)ppGpp chromatographic identification were employed to evaluate the effect of QHD on biofilm in strong biofilm-forming HP strains.The effect of QHD on biofilm and efflux pump-related gene expression was evaluated by quantitative polymerase chain reaction.Non-targeted metabolomics with UHPLC-MS/MS was used to identify potential metabolic pathways and biomarkers which were different between the NC and QHD groups.RESULTS HP could form biofilms of different degrees in vitro,and the intensity of formation was associated with the drug resistance of the strain.QHD had strong bacteriostatic and bactericidal effects on HP,with MICs of 32-64 mg/mL.QHD could inhibit the biofilm formation of the strong biofilm-forming HP strains,disrupt the biofilm structure,lower the accumulation of(p)ppGpp,decrease the expression of biofilm-related genes including LuxS,Spot,glup(HP1174),NapA,and CagE,and reduce the expression of efflux pump-related genes such as HP0605,HP0971,HP1327,and HP1489.Based on metabolomic analysis,QHD induced oxidative stress in HP,enhanced metabolism,and potentially inhibited relevant signaling pathways by upregulating adenosine monophosphate(AMP),thereby affecting HP growth,metabolism,and protein synthesis.CONCLUSION QHD exerts bacteriostatic and bactericidal effects on HP,and reduces HP drug resistance by inhibiting HP biofilm formation,destroying its biofilm structure,inhibiting the expression of biofilm-related genes and efflux pump-related genes,enhancing HP metabolism,and activating AMP in HP.
基金National natural science foundation of China(No.81673905,81873260)National key research and development planning(No.2017YFC1700104)+2 种基金Project of science and technology department of Jiangsu province(No.BE2019769)Project for the construction of superior disciplines of universities and colleges in Jiangsu province[2018(87)]Scientific research innovative program for postgraduate students Jiangsu province(No.KYCX20_1530)。
文摘Objective:To explore the target and signal pathway of Qingchang Huashi Decoction(QCHSD)in the treatment of ulcerative colitis(UC)by using network pharmacology,so as to explain its molecular mechanism of action in the treatment of UC from damp heat.Methods:TCMSP was used to screen the potential active components(OB≥30%,DL≥0.18)and target of QCHSD.The network of"potential active ingredients-target-disease"was constructed by using the database of TCMSP and GeneCards.Using the string platform,the protein protein interaction(PPI)network model was constructed to find the core target.Go and KEGG enrichment of potential targets were analyzed by R software.Results:The results of network analysis showed that quercetin,kaempferol,scutellarin and baicalein were the top four active ingredients in QCHSD.210 gene targets were found in QCHSD,4213 in UC.The key targets of QCHSD in treating UC included AKT1,IL-6,VEGFA,CASP3,etc.GO enrichment analysis showed that these gene targets mainly affected nuclear receptor,steroid receptor,cytokine receptor binding,cytokine activity,etc.KEGG enrichment analysis showed that AGE-RAGE signaling pathway,IL-17 signaling pathway and TNF were more abundant.Conclusion:This study describes the material basis and mechanism of QCHSD in the treatment of UC,which provides theoretical basis and research direction for future research.
