Effects Of Valproate Prenatal Exposure On Hepatocellular Lipid Metabolism In Fetal And Neonatal Mice

Objective: Sodium valproate(sodium valproate,VPA),is an antiepileptic drug characterized with disturbing effect on the body’s metabolism and inducing lipid metabolism disorder in hepatocytes.Based on the potential disturbance of VPA,the effect of prenatal exposure to VPA on lipid metabolism in offspring hepatocytes is not clear.The aim of this study was to construct a prenatal VPA exposure mouse model and to investigate the lipid metabolism of liver cells in fetal and newborn mice through molecular,morphological and functional assays.Methods:(1)Collection and analysis of the clinical data of epilepsy patients taking VPA.(2)To identify the dose of VPA designed,and to establish the prenatal VPA exposure in different embryonic stages of ICR mice.(3)The pregnant mice were randomly divided into three groups: blank control group,a low-concentration VPA group(200 mg/kg),and a high-concentration VPA group(400 mg/kg),with 10 pregnant mice in each group.In order to avoid abortion in mice,the pregnant mice were given with VPA after one week,and the vital signs of the pregnant mice were closely observed during the gavage.Since the day-11.5 of embryonic period(E11.5)is a critical period for liver development,the liver specimens of fetal mice(E11.5)are collected.In addition,the blood of maternal mice,liver tissue samples of maternal,fetal and newborn(PND1)mice were collected.Biochemical indicators,oil red staining,immunohistochemical staining,immunofluorescence staining,and Western blot analysis were performed these specimens.Results: B-ultrasound examination of patients with epilepsy taking VPA were detected with fatty liver,however,there was no significant change in serum glucose and lipid metabolism.The levels of total cholesterol and fibroblast growth factor-21(FGF21)in maternal blood samples were increased significantly after prenatal exposure to VPA.The results of immunostaining and immunoblot analyses showed that the FGF21 and CD36 proteins which promoted lipid uptake in the liver cells of the maternal mice were notably up-regulated,while the expression of endogenous ApoB protein regulated lipid output was decreased.In addition,the levels of TG and FGF21 in the liver homogenate of fetal E11.5 mice and neonatal PND1 mice were markedly increased in prenatal high-dose VPA exposure.Oil red staining showed increased lipid production in the liver tissue of fetal and neonatal mice following prenatal high-dose VPA exposure.FGF21 and CD36 were visibly increased in the liver tissues of fetal and neonatal mice with high prenatal high-dose VPA exposure,while the expression of cellular Apob protein was down-regulated.Conclusion: The antiepileptic drug VPA may induce the risk of lipid metabolism disorder in liver tissue of patients with epilepsy.In addition,prenatal VPA exposure may increase the disrupting effects of lipid metabolism in offspring liver tissue cells,and its molecular mechanism is related to VPA-induced functional disorders of "input" and "output" of lipid substances in hepatocytes.The results of this study can provide a scientific basis for clinicians to guide pregnant women with epilepsy to take VPA drugs with caution.