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BACKGROUND The aim of this study was to investigate the potential anti-obesity effects of Camellia nitidissima Chi flower extract (Cnfe) by examining its effects in terms of the regulation of lipid levels and modulation of gut microbiota in rats with high-fat-diet-induced obesity. RESULTS Our results demonstrated that Cnfe significantly decreased weight gain by reducing appetite and decreasing high-fat food intake. Further, Cnfe restored normal lipid metabolism, and improved insulin sensitivity and glucose tolerance in rats fed a high-fat diet. Real-time RT-PCR and western blot results showed that Cnfe significantly decreased the expression of genes and proteins involved in adipogenesis and lipogenesis, and up-regulated the expression of lipolysis genes. 16S rRNA sequencing of feces showed that Cnfe dramatically reversed dysbacteriosis in rats with high-fat-diet-induced obesity by decreasing the abundance of Firmicutes and increasing that of Bacteroidetes and Verrucomicrobia at the phylum level. CONCLUSION The results demonstrate that Cnfe is a potential anti-obesity prebiotic nutrient that can prevent weight gain, ameliorate obesity-related dyslipidemia and hyperglycemia, inhibit liver fat accumulation, and modulate gut microbiota. This article is protected by copyright. All rights reserved.