{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T10:37:43Z","timestamp":1791110263886,"version":"4.1.0"},"reference-count":47,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T00:00:00Z","timestamp":1708300800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"Background<\/jats:title>Helicobacter pylori<\/jats:italic> (H. pylori<\/jats:italic>) is thought to primarily colonize the human stomach and lead to various gastrointestinal disorders, such as gastritis and gastric cancer. Currently, main eradication treatment is triple or quadruple therapy centered on antibiotics. Due to antibiotic resistance, the eradication rate of H. pylori<\/jats:italic> is decreasing gradually. Therefore, searching for anti-H. pylori<\/jats:italic> drugs from herbal sources has become a strategy for the treatment. Our team proposed a Hezi Qingyou Formula (HZQYF), composed of Chebulae Fructus, Ficus hirta Vahl and Cloves, and studied its anti-H. pylori<\/jats:italic> activity and mechanism.<\/jats:p><\/jats:sec>Methods<\/jats:title>Chemical components of HZQYF were studied using UHPLC\u2013MS\/MS and HPLC. Broth microdilution method and agar dilution method were used to evaluate HZQYF\u2019s antibacterial activity. The effects of HZQYF on expression of adhesion genes (alpA<\/jats:italic>, alpB<\/jats:italic>, babA<\/jats:italic>), urease genes (ureE<\/jats:italic>, ureF<\/jats:italic>), and flagellar genes (flaA, flaB<\/jats:italic>) were explored using Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) technology. Effects on morphology and permeability of the extracellular membrane were studied using scanning electron microscopy (SEM) and N-phenylnaphthalen-1-amine (NPN) uptake. Effect on urease activity was studied using a urease kinetics analysis in vitro<\/jats:italic>. Immunofluorescence staining method was used to examine the effect on adhesion. Western blot was used to examine the effect on cagA protein.<\/jats:p><\/jats:sec>Results<\/jats:title>Minimum inhibitory concentration (MIC) values of the formula against H. pylori<\/jats:italic> clinical strains and standard strains were 80\u2013160\u2009\u03bcg\/mL, and minimum bactericidal concentration (MBC) values were 160\u2013320\u2009\u03bcg\/mL. The formula could down-regulate the expression of adhesion genes (alp<\/jats:italic>A, alpB<\/jats:italic>, babA<\/jats:italic>), urease genes (ureE<\/jats:italic>, ureF<\/jats:italic>) and flagellar genes (flaA<\/jats:italic>, flaB<\/jats:italic>), change the morphology of H. pylori<\/jats:italic>, increase its extracellular membrane permeability, and decrease its urease activity.<\/jats:p><\/jats:sec>Conclusion<\/jats:title>Present studies confirmed that HZQYF had promising in vitro<\/jats:italic> anti-H. pylori<\/jats:italic> activities and demonstrated its possible mechanism of action by down-regulating the bacterial adhesion, urease, and flagellar gene expression, which provided scientific bases for further clinical investigations.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2024.1355460","type":"journal-article","created":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T04:39:26Z","timestamp":1708317566000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":22,"title":["In vitro anti-Helicobacter pylori activity and the underlining mechanism of an empirical herbal formula \u2013 Hezi Qingyou"],"prefix":"10.3389","volume":"15","author":[{"given":"Zhong","family":"Feng","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yajie","family":"Hao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang","family":"Peng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ling","family":"Ou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junwei","family":"Jia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingjin","family":"Xun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanjing","family":"Zou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meiyun","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guimin","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meicun","family":"Yao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,2,19]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1126\/science.1098801","article-title":"Functional adaptation of baba, the h-pylori abo blood group antigen binding adhesin","volume":"305","author":"Aspholm-Hurtig","year":"2004","journal-title":"Science"},{"key":"ref2","volume-title":"Pharmacognostical studies on hakka herbal medicine wuzhimaotao","author":"Au","year":"2009"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"27","DOI":"10.3390\/cells10010027","article-title":"Helicobacter pylori virulence factors-mechanisms of bacterial pathogenicity in the gastric microenvironment","volume":"10","author":"Baj","year":"2020","journal-title":"Cell"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1007\/s00775-013-1002-8","article-title":"Structure of the ured-uref-ureg-uree complex in helicobacter pylori: a model study","volume":"18","author":"Biagi","year":"2013","journal-title":"J. Biol. Inorg. Chem."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"117977","DOI":"10.1016\/j.lfs.2020.117977","article-title":"The gastroprotective potential of silibinin against helicobacter pylori infection and gastric tumor cells","volume":"256","author":"Bittencourt","year":"2020","journal-title":"Life Sci."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"1892","DOI":"10.1126\/science.8018146","article-title":"Attachment of helicobacter pylori to human gastric epithelium mediated by blood group antigens","volume":"262","author":"Boren","year":"1993","journal-title":"Science"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"3071","DOI":"10.3748\/wjg.v24.i28.3071","article-title":"Helicobacter pylori in human health and disease: mechanisms for local gastric and systemic effects","volume":"24","author":"Bravo","year":"2018","journal-title":"World J. Gastroenterol."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1099\/jmm.0.45551-0","article-title":"Role of the helicobacter pylori outer-membrane proteins alpa and alpb in colonization of the guinea pig stomach","volume":"53","author":"de Jonge","year":"","journal-title":"J. Med. Microbiol."},{"key":"ref9","first-page":"A125","article-title":"The role of the helicobacter pylori outer membrane proteins alpa and alpb in the colonisation of the guinea pig stomach","volume":"126","author":"de Jonge","year":"","journal-title":"Gastroenterology"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"e1006464","DOI":"10.1371\/journal.ppat.1006464","article-title":"Helicobacter pylori gene silencing in vivo demonstrates urease is essential for chronic infection","volume":"13","author":"Debowski","year":"2017","journal-title":"PLoS Pathog."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1016\/j.jtcme.2019.05.002","article-title":"Syzygium aromaticum l.: traditional herbal medicine against caga and vaca toxin genes-producing drug resistant helicobacter pylori","volume":"10","author":"El-Shouny","year":"2020","journal-title":"J. Tradit. Complement. Med."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1053\/j.gastro.2019.04.011","article-title":"Reconciliation of recent helicobacter pylori treatment guidelines in a time of increasing resistance to antibiotics","volume":"157","author":"Fallone","year":"2019","journal-title":"Gastroenterology"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"e1001678","DOI":"10.1371\/journal.pbio.1001678","article-title":"Structure of ureg\/uref\/ureh complex reveals how urease accessory proteins facilitate maturation of helicobacter pylori urease","volume":"11","author":"Fong","year":"2013","journal-title":"PLoS Biol."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/j.cgh.2006.09.015","article-title":"Helicobacter pylori baba expression, gastric mucosal injury, and clinical outcome","volume":"5","author":"Fujimoto","year":"2007","journal-title":"Clin. Gastroenterol. Hepatol."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"16847","DOI":"10.1002\/jcp.28363","article-title":"Herbal medicine as an auspicious therapeutic approach for the eradication of helicobacter pylori infection: a concise review","volume":"234","author":"Ghasemian","year":"2019","journal-title":"J. Cell. Physiol."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"773","DOI":"10.1111\/apt.14319","article-title":"Systematic review with meta-analysis: the global recurrence rate of helicobacter pylori","volume":"46","author":"Hu","year":"2017","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"962354","DOI":"10.3389\/fmicb.2022.962354","article-title":"Revealing the novel effect of jinghua weikang capsule against the antibiotic resistance of helicobacter pylori","volume":"13","author":"Jia","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"h4052","DOI":"10.1136\/bmj.h4052","article-title":"Comparative effectiveness and tolerance of treatments for helicobacter pylori: systematic review and network meta-analysis","volume":"351","author":"Li","year":"2015","journal-title":"BMJ"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"e25949","DOI":"10.1097\/MD.0000000000025949","article-title":"Helicobacter pylori reinfection and its risk factors after initial eradication a protocol for systematic review and meta-analysis","volume":"100","author":"Li","year":"2021","journal-title":"Medicine"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"204","DOI":"10.13345\/j.cjb.180239","article-title":"Progress in bacterial urease complexes and their activation mechanisms","volume":"35","author":"Li","year":"2019","journal-title":"Sheng Wu Gong Cheng Xue Bao"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"532","DOI":"10.3760\/cma.j.issn.0578-1426.2017.07.014","article-title":"Fifth chinese national consensus report on the management of helicobacter pylori infection","volume":"56","author":"Liu","year":"2017","journal-title":"Zhonghua Nei Ke Za Zhi"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"6242","DOI":"10.1074\/jbc.M611178200","article-title":"Functional and intracellular signaling differences associated with the helicobacter pylori alpab adhesin from western and east asian strains","volume":"282","author":"Lu","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"5629","DOI":"10.3748\/wjg.v16.i44.5629","article-title":"Screening test for anti-helicobacter pylori activity of traditional chinese herbal medicines","volume":"16","author":"Ma","year":"2010","journal-title":"World J. Gastroenterol."},{"key":"ref24","first-page":"782","article-title":"Study on quality standard of fici hirtae radix","volume":"48","author":"Ma","year":"2017","journal-title":"Chin. Tradit. Herb. Drug"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1136\/gutjnl-2016-312288","article-title":"Management ofhelicobacter pylori infection\u2014the Maastricht v\/Florence consensus report","volume":"66","author":"Malfertheiner","year":"2016","journal-title":"Gut"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"111554","DOI":"10.1016\/j.jinorgbio.2021.111554","article-title":"Probing the transport of ni(ii) ions through the internal tunnels of the helicobacter pylori uredfg multimeric protein complex","volume":"223","author":"Masetti","year":"2021","journal-title":"J. Inorg. Biochem."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"151594","DOI":"10.1016\/j.acthis.2020.151594","article-title":"The validation of the helicobacter pylori caga typing by immunohistochemistry: nationwide application in Indonesia","volume":"122","author":"Miftahussurur","year":"2020","journal-title":"Acta Histochem."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"4081","DOI":"10.1128\/jb.179.13.4081-4086.1997","article-title":"Characterization of ureg, identification of a ured-uref-ureg complex, and evidence suggesting that a nucleotide-binding site in ureg is required for in vivo metallocenter assembly of klebsiella aerogenes urease","volume":"179","author":"Moncrief","year":"1997","journal-title":"J. Bacteriol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1186\/s13099-016-0091-7","article-title":"Systematic review of time trends in the prevalence of helicobacter pylori infection in China and the USA","volume":"8","author":"Nagy","year":"2016","journal-title":"Gut Pathogens"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1136\/gut.41.1.37","article-title":"Blood groups Lewis(b) and abh expression in gastric mucosa: lack of inter-relation with helicobacter pylori colonisation and occurrence of gastric malt lymphoma","volume":"41","author":"Oberhuber","year":"1997","journal-title":"Gut"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"1537","DOI":"10.1046\/j.1365-2958.1999.01300.x","article-title":"Genetic and functional characterization of the alpab gene locus essential for the adhesion of helicobacter pylori to human gastric tissue","volume":"31","author":"Odenbreit","year":"1999","journal-title":"Mol. Microbiol."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"117428","DOI":"10.1016\/j.jep.2023.117428","article-title":"Terminalia chebula retz. Aqueous extract inhibits the helicobacter pylori-induced inflammatory response by regulating the inflammasome signaling and er-stress pathway","volume":"320","author":"Ou","year":"2023","journal-title":"J. Ethnopharmacol."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"105626","DOI":"10.1016\/j.jff.2023.105626","article-title":"Syzygium aromaticum enhances innate immunity by triggering macrophage m1 polarization and alleviates helicobacter pylori-induced inflammation","volume":"107","author":"Peng","year":"2023","journal-title":"J. Funct. Food."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"114995","DOI":"10.1016\/j.jep.2022.114995","article-title":"In vitro anti-helicobacter pylori activity of syzygium aromaticum and the preliminary mechanism of action","volume":"288","author":"Peng","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1186\/s13020-021-00442-1","article-title":"In vitro anti-bacterial activity and network pharmacology analysis of sanguisorba officinalis l. against helicobacter pylori infection","volume":"16","author":"Shen","year":"2021","journal-title":"Chin. Med."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1089\/dna.2018.4487","article-title":"Helicobacter pylori-induced dna damage is a potential driver for human gastric cancer ags cells","volume":"38","author":"Shi","year":"2019","journal-title":"DNA Cell Biol."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"310","DOI":"10.3949\/ccjm.84a.14110","article-title":"Treating helicobacter pylori effectively while minimizing misuse of antibiotics","volume":"84","author":"Shiotani","year":"2017","journal-title":"Clevel. Clin. J. Med."},{"key":"ref38","first-page":"734","article-title":"Influence of polygonum capitatum on the adherency and colonization of helicobacter pylori","volume":"8","author":"Shu","year":"2016","journal-title":"Chinese J. Zoonoses"},{"key":"ref39","doi-asserted-by":"publisher","first-page":"49359","DOI":"10.1074\/jbc.M108619200","article-title":"Crystal structure of klebsiella aerogenes uree, a nickel-binding metallochaperone for urease activation","volume":"276","author":"Song","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"514","DOI":"10.1111\/apt.13497","article-title":"Review article: the global emergence of helicobacter pylori antibiotic resistance","volume":"43","author":"Thung","year":"2016","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"1119037","DOI":"10.3389\/fcimb.2023.1119037","article-title":"A new approach to overcoming antibiotic-resistant bacteria: traditional chinese medicine therapy based on the gut microbiota","volume":"13","author":"Xue","year":"2023","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref43","doi-asserted-by":"publisher","first-page":"114578","DOI":"10.1016\/j.jep.2021.114578","article-title":"In-vitro anti-helicobacter pylori activity and preliminary mechanism of action of canarium album raeusch. Fruit extracts","volume":"283","author":"Yan","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"1015","DOI":"10.1007\/s00011-021-01501-x","article-title":"Chronic inflammation and long-lasting changes in the gastric mucosa after helicobacter pylori infection involved in gastric cancer","volume":"70","author":"Yang","year":"2021","journal-title":"Inflamm. Res."},{"key":"ref45","first-page":"118","article-title":"Critical role of alpa and alpb for biofilm formation and cell adhesion of helicobacter pylori","volume":"21","author":"Yonezawa","year":"2016","journal-title":"Helicobacter"},{"key":"ref46","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1016\/j.jep.2012.03.001","article-title":"Anti-inflammatory and cytoprotective effects of selected pakistani medicinal plants in helicobacter pylori-infected gastric epithelial cells","volume":"141","author":"Zaidi","year":"2012","journal-title":"J. Ethnopharmacol."},{"key":"ref47","doi-asserted-by":"publisher","first-page":"743","DOI":"10.1016\/j.etp.2011.01.009","article-title":"Effects of ficus hirta vahl. (Wuzhimaotao) extracts on growth inhibition of hela cells","volume":"64","author":"Zeng","year":"2012","journal-title":"Exp. Toxicol. Pathol."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"e36844","DOI":"10.1371\/journal.pone.0036844","article-title":"Risk assessment of gastric cancer caused by helicobacter pylori using caga sequence markers","volume":"7","author":"Zhang","year":"2012","journal-title":"PLoS One"}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2024.1355460\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T04:39:47Z","timestamp":1708317587000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2024.1355460\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,19]]},"references-count":47,"alternative-id":["10.3389\/fmicb.2024.1355460"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2024.1355460","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,19]]},"article-number":"1355460"}}