Rat liver, rat liver cells (BRL cells) and mouse macrophages (ANA-1 cells) were used to investigate the effects of alfalfa saponins (AS) on mRNA expressions of reverse cholesterol transport genes, which were ATP-binding cassette transporter A1 (ABCA1) and scavenger receptor class B type Ⅰ(SR-BⅠ), and discuss the effects of AS from animal and cellular levels. Thirty two healthy male SD rats were randomly divided into four groups:normal control group, AS group, hyperlipidemic model group and hyperlipidemic saponins group, and each group had 8 rats. Rats in normal control group and AS group were fed a basal diet, and those in the other two groups were fed a high-fat diet. After 4 weeks feeding, AS[240 mg/(kg·d)] was intragastric administered to rats in AS group and hyperlipidemic saponins group from weeks 5 to 8. BRL cells were incubated with fetal bovine serum for 48 h to constitute hyperlipidemic model. BRL cells were divided into four groups, which were normal control group (normal cells), AS group (normal cells), hyperlipidemic model group (hyperlipidemic cells) and hyperlipidemic saponins group (hyperlipidemic cells), and AS (final concentration 300 μg/mL) was added to culture medium in AS group and hyperlipidemic saponins group to culture for 24 h. ANA-1 cells were incubated with oxidized low density lipoprotein for 48 h to constitute lipid-loaded model. ANA-1 cells were divided into four groups, which were normal control group (normal cells), AS group (normal cells), lipid-loaded model group (lipid-loaded cells) and lipid-loaded saponins group (lipid-loaded cells), and AS (final concentration 300 μg/mL) was added to culture medium in AS group and lipid-loaded saponins group to culture for 24 h. The mRNA expressions of ABCA1 and SR-BⅠ in rat liver, BRL cells and ANA-1 cells were determined by fluorescent quantitative PCR. The results showed as follows:1) AS significantly increased the mRNA expressions of ABCA1 and SR-BⅠ in liver of normal rats and of ABCA1 in liver of hyperlipidemic rats (P<0.05), but had no significant effects on that of SR-BⅠ in liver of hyperlipidemic rats (P>0.05); 2)AS significantly increased the mRNA expressions of ABCA1 and SR-BⅠ in normal BRL cells(P<0.05), but had no significant effects on those in hyperlipidemic BRL cells (P>0.05); 3) AS significantly reduced the mRNA expression of ABCA1 in normal ANA-1 cells (P<0.05). In conclusion, AS might promote reverse cholesterol transport by up-regulating the mRNA expressions of ABCA1 and SR-BⅠin rat liver and normal BRL cells, promote the excretion of liver cholesterol, and play prevention and curing effects on hyperlipidemia.