This experiment was conducted to study the effects of dietary restriction on cholesterol metabolism in liver and adrenal gland of mice. ICR male mice were used as the study subjects, with adaptive feeding 3 days. After weighing up to 24 to 25 g, they were randomly divided into 5 groups with 6 mice in each group, including normal control group (free feeding), 4.0 g feeding group[feed intake:4.0 g/d per mouse; restriction ratio:88% of normal feed intake], 3.0 g feeding group[feed intake:3.0 g/d per mouse; restriction ratio:66% of normal feed intake], 2.0 g feeding group[feed intake:2.0 g/d per mouse; restriction ratio:44% of normal feed intake] and 1.5 g feeding group[feed intake:1.5 g/d per mouse; restriction ratio:22% of normal feed intake]. It was free drinking for all groups. After feeding restriction for 14 days, the mice were executed, and the heart, liver, kidney, spleen, thymus and testis were weighed and the organic indexes were calculated. The total RNA and total protein of hepatic and adrenal tissues were extracted. Then, cholesterol metabolism related genes mRNA relative expression levels were detected by real-time quantitative PCR technology, and steroidogenic acute regulatory protein (StAR) relative expression level was assayed by Western blot method, and serum corticosterone content were tested by enzyme-linked immuno sorbent assay (ELISA) method. The results showed as follows:1) compared with the normal control group, the body weight of mice in dietary restriction groups was lost in different degrees, and the heart, liver, kidney, spleen and thymus of mice in dietary restriction groups were shrunken in different degrees. 2) Compared with the normal control group, the expression of low-density lipoprotein receptor (Ldlr), scavenger receptor class B member 1 (Scarb1), hydroxymethyl glutaric acyl coenzyme A reductase (Hmgcr), hormone-sensitive triglayceride lipase (Lipe), apolipoprotein E (Apoe), ATP-binding cassette transporter G5 (Abcg5), ATP-binding cassette transporter G8 (Abcg8), cytochromes P450 7A1 (Cyp7a1), sterol regulatory element binding protein cleavage active protein (Scap), liver X receptor β (Nr1h2), peroxisome proliferators-activated receptor α (Ppara) and peroxisome proliferators-activated receptor γ (Pparg) genes in liver of 3.0 g feeding group, 2.0 g feeding group and 1.5 g feeding group were significantly up-regulated (P<0.05 or P<0.01), while the expression of insulin induced gene 2 (Insig2) gene was significantly down-regulated (P<0.05). 3) Compared with the normal control group, the expression of adrenal cytochromes P450 11A1 (Cyp11a1) and cytochromes P450 21A1 (Cyp21a1) genes in adrenal gland of 2.0 g feeding group and 1.5 g feeding group were significantly up-regulated (P<0.05), the expression of StAR and cytochromes P450 11B1 (Cyp11b1) genes in adrenal gland of 1.5 g feeding group were significantly up-regulated (P<0.05), and the expression of cytochromes P450 11B2 (Cyp11b2) in 3.0 g feeding group, 2.0 g feeding group and 1.5 g feeding group was significantly down-regulated (P<0.05). Additionally, the adrenal StAR protein expression of all dietary restriction groups was strengthen, and it was strengthen with the feed intake decreasing. In conclusion, the organism metabolism such as spleen, thymus and liver are affected by extreme dietary restriction, and then the hepatic and adrenal cholesterol metabolism and conversion are affected.