Objective To establish a liver-specific Rbp4 gene knockout mouse model and to explore the effect of liver Rbp4 gene deletion on glucose metabolism. Methods Cre-LoxP technology was used to construct a liver-specific Rbp4 gene knockout mouse model using C57 / BL6J and Alb-Cre mice. The genotype of the mice was identified by polymerase chain reaction and agarose gel electrophoresis. Ten 18 week old C57 / BL6J male mice were included in the WT group, 10 flox homozygous and Alb-Cre negative mice of the same age were included in the experimental control group(Rbp4 flox / flox: Cre-), and 10 flox homozygous and Alb-Cre positive mice of the same age were included in the experimental group (Rbp4 flox / flox: Cr e+). Expression levels of RBP4 protein and mRNA in the liver were verified by Western Blot and quantitative reverse transcription-polymerase chain reaction ( qRT-PCR), respectively, and expression levels of Rbp4 mRNA in other tissues were detected by qRT-PCR. Morphological changes in liver tissue were detected by hematoxylin and eosin staining. Blood glucose values were detected in mouse tail vein blood samples using a blood glucose meter, and glucose tolerance and insulin tolerance were determined. Expression levels of the liver glucose metabolism genes phosphoenolpyruvate carboxylase ( Pepck) and glucose-6-phosphatase ( G6pase) were detected by qRT-PCR. Results Liver-specific Rbp4 knockout mice were successfully bred and identified. RBP4 protein and mRNA levels were significantly decreased in the liver of Rbp4flox / flox: Cre + mice ( P< 0. 05), but there was no significant difference in the relative expression levels of Rbp4 mRNA in fat, kidney, pancreas, spleen, heart, or muscle tissues among the three groups (P>0. 05). Liver-specific Rbp4 knockout had no significant effect on liver morphology, glucose tolerance, or insulin tolerance (P> 0. 05). Pepck mRNA levels in the liver differed significantly among the three groups (P< 0. 05), and pairwise comparison showed that liver Pepck mRNA levels were significantly lower in Rbp4 flox / flox: Cre + mice compared with levels in Rbp4 flox / flox: Cre - mice (P< 0. 05). There was no significant difference in liver glucose-6-phosphatase (G6pase) mRNA expression among the three groups (P> 0. 05). Conclusions We successfully constructed a liver-specific Rbp4 knockout mouse model. Deletion of Rbp4 in the liver inhibited expression of Pepck mRNA in the liver, thus providing a basis for further exploration of the role of this gene in glucose metabolism in mice.