Helicobacter　pylori　(H.　pylori)　is　a　major　factor　in　peptic　ulcer　disease　and　gastric　cancer,　and　its　infection　rate　is　rising　globally.　The　efficacy　of　traditional　antibiotic　treatment　is　less　effective,　mainly　due　to　bacterial　biofilms　and　the　formation　of　antibiotic　resistance.　In　addition,　H.　pylori　colonizes　the　gastrointestinal　epithelium　covered　by　mucus　layers,　the　drug　must　penetrate　the　double　barrier　of　mucus　layer　and　biofilm　to　reach　the　infection　site　and　kill　H.　pylori.　The　ethanol　injection　method　was　used　to　synthesize　nanoliposomes　(EPI/R-AgNPs@RHL/PC)　with　a　mixed　lipid　layer　containing　rhamnolipids　(RHL)　and　phosphatidylcholine　(PC)　as　a　carrier,　loaded　with　the　urease　inhibitor　epiberberine　(EPI)　and　the　antimicrobial　agent　rubropunctatin　silver　nanoparticles　(R-AgNPs).　EPI/R-AgNPs@RHL/PC　had　the　appropriate　size,　negative　charge,　and　acid　sensitivity　to　penetrate　mucin-rich　mucus　layers　and　achieve　acid-responsive　drug　release.　In　vitro　experiments　demonstrated　that　EPI/R-AgNPs@RHL/PC　exhibited　good　antibacterial　activity,　effectively　inhibited　urease　activity,　removed　the　mature　H.　pylori　biofilm,　and　inhibited　biofilm　regeneration.　In　vivo　antibacterial　tests　showed　that　EPI/R-AgNPs@RHL/PC　exhibited　excellent　activity　in　eradicating　H.　pylori　and　protecting　the　mucosa　compared　to　the　traditional　clinical　triple　therapy,　providing　a　new　idea　for　the　treatment　of　H.　pylori　infection.　©　2023　Elsevier　B.V.