Abstract: Objective To study the inhibitory effect of valproate (VPA) on rifampin-dependent tubercle bacilli. Methods BALB/c mice were infected with rifampin-dependent tubercle bacilli strains, rifampin-resistant tubercle bacilli strains and H37Rv, respectively. Six weeks after they were treated with combined rifampin and VPA, their physical signs were observed, and severity of lung and spleen lesions, lung and spleen weight indexes, tubercle bacilli strains load in lung tissues, and rifampin-dependent tubercle bacilli strains were detected. Results The lesions were severer, the lung and spleen weight indexes and the tubercle bacilli strains load in lung tissues were higher in rifampin-dependent tubercle bacilli strains infection mouse model and rifampin treatment group.Combined rifampin and VPA could effectively reduce the tubercle bacilli strains load in lung tissues of mice. Conclusion VPA can significantly inhibit rifampin-dependent tubercle bacilli in mice.