BanXiaXieXin decoction treating gastritis mice with drug-resistant Helicobacter pylori and its mechanism

BACKGROUND Helicobacter pylori(H.pylori)is the main pathogen that causes a variety of upper digestive diseases.The drug resistance rate of H.pylori is increasingly higher,and the eradication rate is increasingly lower.The antimicrobial resistance of H.pylori is an urgent global problem.It has been confirmed that Banxia Xiexin decoction(BXXXT)demonstrates the effects of treating gastrointestinal diseases,inhibiting H.pylori and protecting gastric mucosa.The purpose of the present study is to further explore the therapeutic effects of BXXXT on drug-resistant H.pylori.AIM To confirm that BXXXT demonstrates therapeutical effects in vivo and in vitro on gastritis mice with drug-resistant H.pylori and explain its mechanism to provide an experimental basis for promoting the application of BXXXT.METHODS The aqueous extract of BXXXT was gained by water decocting method.The inhibitory effect of the aqueous extract on H.pylori was detected by dilution in vitro;drug-resistant H.pylori cells were used to build an acute gastritis model in vivo.Thereafter,the model mice were treated with the aqueous extract of BXXXT.The amount of H.pylori colonization,the repair of gastric mucosal damage,changes of inflammatory factors,apoptosis,etc.,were assessed.In terms of mechanism exploration,the main medicinal compositions of BXXXT aqueous extract and the synergistic bacteriostatic effects they had demonstrated were analyzed using mass spectrometry;the immune function of peripheral blood cells such as CD3+T and CD4+T of mice with gastritis before and after treatment with BXXXT aqueous extract was detected using a flow cytometry;the H.pylori transcriptome and proteome after treatment with BXXXT aqueous extract were detected.Differently expressed genes were screened and verification was performed thereon with knockout expression.RESULTS The minimum inhibitory concentration of BXXXT aqueous extract against H.pylori was 256-512μg/mL.A dose of 28 mg/kg BXXXT aqueous extract treatment produced better therapeutical effects than the standard triple therapy did;the BXXXT aqueous extract have at least 11 ingredients inhibiting H.pylori,including berberine,quercetin,baicalin,luteolin,gallic acid,rosmarinic acid,aloe emodin,etc.,of which berberine,aloe emodin,luteolin and gallic acid have a synergistic effect;BXXXT aqueous extract was found to stimulate the expressions of CD3+T and CD4+T and increase the number of CD4+T/CD8+T in gastritis mice;the detection of transcriptome and proteome,quantitative polymerase chain reaction,Western blotting and knockout verification revealed that the main targets of BXXXT aqueous extract are CFAs related to urea enzymes,and CagA,VacA,etc.CONCLUSION BXXXT aqueous extract could demonstrate good therapeutic effects on drug-resistance H.pylori in vitro and in vivo and its mechanism comes down to the synergistic or additional antibacterial effects of berberine,emodin and luteolin,the main components of the extract;the extract could activate the immune function and enhance bactericidal effects;BXXXT aqueous extract,with main targets of BXXXT aqueous extract related to urease,virulence factors,etc.,could reduce the urease and virulence of H.pylori,weaken its colonization,and reduce its inflammatory damage to the gastric mucosa.

Linolenic acid-metronidazole inhibits the growth of Helicobacter pylori through oxidation

BACKGROUND Resistance to antibiotics is one the main factors constraining the treatment and control of Helicobacter pylori(H.pylori)infections.Therefore,there is an urgent need to develop new antimicrobial agents to replace antibiotics.Our previous study found that linolenic acid-metronidazole(Lla-Met)has a good antibacterial effect against H.pylori,both antibiotic-resistant and sensitive H.pylori.Also,H.pylori does not develop resistance to Lla-Met.Therefore,it could be used for preparing broad-spectrum antibacterial agents.However,since the antibacterial mechanism of Lla-Met is not well understood,we explored this phenomenon in the present study.AIM To understand the antimicrobial effect of Lla-Met and how this could be applied in treating corresponding infections.METHODS H.pylori cells were treated with the Lla-Met compound,and the effect of the compound on the cell morphology,cell membrane permeability,and oxidation of the bacteria cell was assessed.Meanwhile,the differently expressed genes in H.pylori in response to Lla-Met treatment were identified.RESULTS Lla-Met treatment induced several changes in H.pylori cells,including roughening and swelling.In vivo experiments revealed that Lla-Met induced oxidation,DNA fragmentation,and phosphatidylserine ectropionation in H.pylori cells.Inhibiting Lla-Met with L-cysteine abrogated the above phenomena.Transcriptome analysis revealed that Lla-Met treatment up-regulated the expression of superoxide dismutase SodB and MdaB genes,both anti-oxidation-related genes.CONCLUSION Lla-Met kills H.pylori mainly by inducing oxidative stress,DNA damage,phosphatidylserine ectropionation,and changes on cell morphology.