Objective To explore method for improving the colonization efficiency of Helicobacter pylori (Hp) in the mouse stomach and to determine if the proton pump inhibitor(PPI) pantoprazole can act as a colonization adjuvant to enhance Hp colonization, with the aim of providing an effective tool for establishing an Hp infection mouse model. Methods The Hp Sydney strain 1(SS1) was introduced and solid plate and liquid culture systems were established.The effects of different doses of pantoprazole on gastric acid secretion in mice were compared. The impact of Hp inoculation, alone or combined with pantoprazole pretreatment, on Hp colonization efficiency was analyzed using rapid urease tests, bacterial plate cultures, and TaqMan quantitative polymerase chain reaction. Results PPI pretreatment inhibited gastric acid secretion and promoted Hp colonization in the mouse stomach, to some extent. Conclusions PPI can serve as colonization adjuvants to enhance the efficiency of constructing Hp infection mouse models.