AimWe sought to profile gut microbiota's role in combination of Bu Fei Hua Yu (BFHY) with cisplatin treatment.MethodsNon-small cell lung cancer (NSCLC) mice model were constructed followed by treatment with cisplatin alone or combined with BFHY. Mice weight and tumor volume were measured during the experiment. And mice cecum were detected by hematoxylin and eosin, cecum contents were collected for Enzyme Linked ImmuneSorbent Assay, and stool were profiled for metagenomic sequencing.ResultsCombination of BFHY with cisplatin treatment decreased the tumor growth and relieved the damage of cecum. Expressions of interleukin-6 (IL-6), interleukin-1β(IL-1β), monocyte chemotactic protein 1 (MCP), and interferon-γ(IFN-γ) were decreased compared with cisplatin treatment alone. Linear discriminant analysis effect size analysis showed thatg_Parabacteroideswas downregulated andg_Escherichiaandg_Blautiawere upregulated after cisplatin treatment. After combination with BFHY,g_Bacteroidesandg_Helicobacterwere decreased.g_Klebsiella,g_Unclssified_Proteobacteria, andg_Unclssified_Clostridiateswere increased. Moreover, heatmap results showed thatBacteroidesabundance was increased significantly after cisplatin treatment; BFHY combination treatment reversed this state. Function analysis revealed that multiple functions were slightly decreased in cisplatin treatment alone and increased significantly after combination with BFHY.ConclusionOur study provided evidence of an efficacy of combination of BFHY with cisplatin on treatment of NSCLC and revealed that gut microbiota plays a role in it. The above results provide new ideas on NSCLC treatment.