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This study aimed to elucidate the underlying mechanisms for the preventive effect of procyanidin B2 (PB2) on a high-fat diet (HFD)-induced dyslipidemia in C57BL/6 mice through a comprehensive analysis of fecal gut microbiome and plasma metabolomics. PB2 significantly reduced the levels of lipid metabolism biomarkers and increased the antioxidant abilities and lipoprotein lipase activity in mice fed with HFD (P < 0.05). Furthermore, the abundance of Blautia genus was increased, and that of Lachnoclostridium, Clostridium, Bilophila, Proteus genera were decreased in the PB2 group of mice compared to the HFD group of mice. Additionally, purine, unsaturated fatty acids, glycerophospholipids, and bile acids were up-regulated in the HFD fed mice plus PB2, and phenylglucuronide was down-regulated. The correlation analysis indicated that the gut microbiome, particularly Bilophila and Proteus genera, were strongly associated with the metabolic pathway modulation, which might have led to the preventive effect of PB2 on dyslipidemia.