Objective To investigate the long-term effects of maternal daidzein intervention on lipid metabolism in offspring mice with gestational diabetes and its mechanism of action. Methods Thirty C57BL/6J female mice were randomly divided into normal, diabetic and daidzein groups, with 10 mice in each group. After the mice in the diabetes group and daidzein group were fed a high-fat and high-sugar diet for 4 weeks, the male and female mice were housed together and streptozotocin (65 mg/kg) was injected intraperitoneally to induce a gestational diabetes model. Starting from the gestation period of the mother mice, the normal group and the diabetic group were given normal saline by gavage (0.5 mL), and the daidzein group was given daidzein (100 mg/kg/d) by gavage until the mice were weaned (3 weeks old). Ten male and ten female offspring mice were retained in each group, and the male and female offspring were kept in separate cages until 20 weeks of age. The serum and liver of the offspring mice were collected to detect the expression of blood glucose, blood lipids and lipid metabolism-related proteins(SREBP-1c, FAS, PPARα, CPT-1, ACOX1). Results Compared with the diabetic group, daidzein intervention improved the metabolic indexes of maternal mice. In terms of offspring, the daidzein group had lower body weight, lower blood sugar, and higher insulin than the diabetic offspring mice (P<0.05). Serum levels of TG, TC and LDL-C were significantly lower and HDL-C levels were significantly higher in both female and offspring mice(P<0.05).AST and ALT levels were significantly lower (P<0.05). Hepatic fat vacuoles and cellular balloon-like degeneration significantly reduced. The expression of SREBP-1c and FAS decreased, and the expression of PPARα, CPT-1 and ACOX1 elevated(P<0.05). The indexes of daidzein offspring female mice were significantly better than those of male mice(P<0.05), and there was no significant difference compared with normal offspring (female and male) mice (P>0.05). Conclusion Maternal daidzein intervention may improve lipid metabolism disorders in offspring of gestational diabetic mice by regulating the SREBP-1c/FAS and PPARα/CPT-1/ACOX1 signaling pathways, and the protective effect on female offspring is better than that on males