To investigate the clinical effect of CO2 fractional laser combined with tacrolimus in the treatment of vitiligo. Methods A total of 94 patients with vitiligo admitted to our hospital from January 2020 to October 2023 were selected as the research objects. They were divided into control group and observation group by simple sampling method, with 47 patients in each group. The control group was treated with tacrolimus ointment, and the observation group was treated with CO2 fractional laser on the basis of the control group. The vitiligo area severity index (VASI), vitiligo disease activity score (VIDA), skin lesion distribution score, inflammatory factor levels (TNF-α, ICAM-1, IL-17, TGF-β1) and the incidence of adverse reactions were compared between the two groups. Results After 4 weeks of intervention, the VASI, VIDA and skin lesion distribution scores of the observation group were lower than those of the control group (P <0.05). The levels of TNF-α, ICAM-1, IL-17 and TGF-β1 in the observation group were lower than those in the control group after 4 weeks of intervention (P <0.05). There was no significant difference in the incidence of adverse reactions between the two groups (P >0.05). Conclusion CO2 fractional laser combined with tacrolimus is effective in the treatment of vitiligo, which can reduce the clinical symptoms and skin lesion distribution, reduce the inflammatory response in vivo, and has high safety.