Antimicrobial　resistance　has　been　considered　as　a　great　threat　to　biosecurity　and　human　health.　And　the　transmission　of　antibiotic　resistance　genes　(ARGs)　by　conjugated　plasmid　is　a　key　factor　in　the　prevalence　of　antimicrobial　resistance.　Paracetamol　(PRC),　one　of　nonopioid　analgesics,　is　an　extensively　used　antipyretic　and　mild　analgesic　worldwide　available　for　numerous　prescriptions.　It　was　unclear　whether　PRC　could　promote　the　spread　of　ARGs.　Here,　it　was　demonstrated　that　PRC　promoted　intergenera　conjugative　plasmid　transfer　in　an　established　conjugation　model.　Both　donor　and　recipient　strains　treated　by　PRC　emerged　the　variations　of　reactive　oxygen　species　(ROS),　SOS　response　and　cell　membrane　permeability.　Correspondingly,　transcriptome　analysis　revealed　that　the　gene　expression　involved　in　cell　membrane　permeability　and　SOS　response　was　upregulated　significantly　after　PRC　exposure.　More　directly,　PRC　also　increased　the　expressions　of　conjugation　related　genes　of　trbG　and　trbP　in　donor.　This　study　proved　for　the　first　time　that　PRC　could　enhance　the　intergenera　conjugative　plasmid　transfer.　Collectively,　these　findings　manifested　the　potential　threat　associated　with　the　existence　of　non-antibiotic　substance　PRC,　which　could　provide　an　important　insight　into　antimicrobial　resistance　spread.