〔Abstract〕 Objective To construct hepatocyte-specific silence information regulator 3(Sirt3) gene knockout(Sirt3Δhep) mice by Cre-loxP technique,and to provide an important animal model for further studying the biological function of the hepatocyte Sirt3 gene in diseases.Methods LoxP-labeled Sirt3flox/floxmice were mated with Alb-Cre homozygous(Alb-Cre+/+) mice,and the F1 generation Sirt3flox/-/Alb-Cre+/-mice were then mated with Sirt3flox/floxmice,and the F2 genotype of Sirt3flox/flox/Alb-Cre+/-mice were the Sirt3Δhepmice constructed in this experiment.Sirt3flox/flox/Alb-Cre-/-(Sirt3flox/flox) mice were the control mice.Mouse tail genome DNA was extracted and PCR was used to identify the genotypes of the offspring mice.Immunofluorescence was used to detect Sirt3 expression in mouse hepatocytes.Primary hepatocytes and tissue proteins of Sirt3Δhepmice were extracted,and the expression of Sirt3 in mouse hepatocytes and other tissues was verified by Western blot.HE staining was used to observe mice's liver,heart,spleen,and lung tissue structure.Results Sirt3Δhepmice were successfully identified.Immunofluorescence and Western blot results demonstrated a significant decrease in the expression of Sirt3 in the hepatocytes of these mice compared to the control group(P<0.01).At the same time,there was no significant difference in the expression of Sirt3 in the heart,spleen,kidney,and lung tissues of Sirt3Δhepmice compared with the control group(P>0.05).The results of HE staining showed that the histological characteristics of the liver,heart,spleen,lungs,kidneys,and other major organs of Sirt3Δhepmice were not significantly different from those of the control group mice.Conclusion Hepatocyte-specific Sirt3 gene knockout mice are successfully constructed,which provides an animal model to explore further the role and molecular mechanism of the hepatocyte Sirt3 gene in diseases.