Objective To explore the expressions and interaction of P32 and PKD1 during the genesis and development of pulmonary fibrosis in rats, and expression variation of the two factors after the intervention of active vitamin D3. Methods Ninety male SD rats were randomly divided into model group, treatment group and control group (30 in each group). Bleomycin (BLM, 5 mg/kg) was injected into the trachea of rats to establish the model of pulmonary fibrosis in the model group and the treatment group, while the control group was intratracheally injected with isopyknic saline (500 μl/kg). From the next day of BLM injection, the rats of the treatment group were intraperitoneally injected with 1,25-(OH)2D3 (2 μg/kg) every other day, while the rats of the model group and the control group were intraperitoneally injected with isopyknic VD3 solvent (99.9% propylene glycol and 0.1% ethanol) and saline respectively at the same time. On the 14th, 21st and 28th day after BLM injection, 10 rats in each group were sacrificed. The pulmonary fibrosis was verified by measuring the content of hydroxyproline in the lungs of the rats. The mRNA and protein expression levels of P32 and PKD1 were detected by RT-PCR and immunohistochemistry respectively. Results At the early stage (the 14th day) of pulmonary fibrosis, the expressions of P32 and PKD1 in the treatment group were higher than those in the model group at both mRNA and protein levels (P < 0.05). Correlation analysis showed that the expressions of these two genes had significant correlations at both mRNA and protein levels. Conclusions Active VD3 may promote the the expressions of P32 and PKD1 at early stage during pulmonary fibrosis in rats. Active VD3 can achieve antioxidant effect through the interaction between P32 and PKD1 in the mitochondria, thereby inhibit the development of pulmonary fibrosis.