Objective This study aims to investigate the effects of high-fat diet rich in perilla oil on the expression of key genes that regulate hepatic VLDL synthesis in obese rats. Methods Sixty healthy male 5-week old SD rats were randomly divided into 2 groups. The rats in the normal control group (NC, n=12) were given normal diet, and the rats in the high fat group (HF, n=48) were given a pure high fat diet in order to induce rat models of obesity. In the intervention period, the obesity model rats were randomly divided into 4 subgroups including consistent high fat group (CHF) and three intervention groups depending on perilla oil substitution rate of lard in CHF: 20%PO, 50%PO and 100%PO. The serum triglyceride (TG) of the rats was measured after 4 weeks. Real-time PCR was applied to measure microsomal triglyceride transfer protein (Mtp) and apolipoprotein B (Apob) mRNA, and western blot assay was used for detecting the expression of MTP and APOB in the liver. Results Compared with the NC group, the CHF rats exhibited significantly high fat deposition. The serum TG was markedly higher and the MTP and APOB were decreased at gene and protein levels in the CHF group compared with the NC group. After the intervention, PO remarkably reduced the level of serum TG and decreased hepatic fat deposition as it showed by pathological examination. At the gene and protein levels, MTP and APOB were upregulated by PO to different degrees. Conclusions All the three PO intervention can promote VLDL synthesis and secretion, and decrease the hepatic fat deposition in the obese rats. Furthermore, PO upregulates the expression of MTP at gene and protein levels in a dose-dependent manner.