In this study, a microcosm experiment for 120 days exposure of microplastics (MPs) on cold seep sediment was conducted. The results showed that different types and doses of MPs addition negatively affected the anaerobic oxidation of CH4. The CH4 oxidation rates with 0.05 % PA and PET addition were only account for approximately 33% of that for the biotic control group. MPs addition did not significantly influence the archaeal community structures but caused a redistribution of bacterial community compositions. The addition of PA, PE, and PET could significantly increase the relative abundance of Desulfobacterota and made Caldatribacteriota almost undetectable. By the present study, we can have a preliminary understanding of the effects of microplastics on anaerobic oxidation of CH4 and prokaryotic communities in the cold seep sediment.