To investigate the effect of berberine regulation of the nuclear factor κB(NF-κB) signaling pathway on human gastric cancer cells infected with Helicobacter pylori(Hp).Methods:The Hp-infected human gastric cancer cell model was established and the cells were divided into a control group(normal saline)，a berberine group(100 μg/mL)，a 5-fluorouracil(5-Fu) group(5 μg/mL)，and a combination group(100 μg/mL berberine+5 μg/mL 5-Fu).The proliferation，cell cycles，protein expression related to the NF-κB signaling pathway，and inflammatory response index levels of Hp-infected human gastric cancer cells in the four groups were compared.Results:The proliferation rates of gastric cancer cells in all groups showed a decreasing trend on the 2nd day and 3rd day of the intervention.The Hp-infected human gastric cancer cells in the G0/G1 and S phases in all groups showed a gradually decreasing trend，while those in the G2/M phase showed a gradually increasing trend.The expression level of IκBα protein in all groups showed a gradually increasing trend，while the expression level of P65 protein showed a gradually decreasing trend.The level of IL-4 in all groups showed a gradually increasing trend，while the level of IL-6 showed a gradually decreasing trend.There were significant differences among all groups(all P<0.05).Conclusion:Berberine inhibits the proliferation and inflammatory response of Hp-infected human gastric cancer cells by regulating the expression of proteins related to the NF-κB signaling pathway，which is worthy of clinical attention.