Abstract: Rat primary cultured hepatocytes were isolated by two-step collagenase digestion method and treated with gradient concentration of SU-MIAN-XIN and BAO-DING-NING, and the expression of cytochrome P450 3A1 (CYP3A1) and cytochrome P450 2E1 (CYP 2E1) were determined by Western blot. The results as follows: 2-4×108 hepatocytes per rat were obtained and the viability was about 95%. The expression of CYP3A1 and CYP 2E1 in rat hepatocytes increased gradually with treatment of SU-MIAN-XIN and BAO-DING-NING. Rat primary cultured hepatocytes can be used for molecular mechanism of SU-MIAN-XIN metabolism, it provides a foundation for its clinical application.