Effects of Zigui yichong formula on premature ovarian insufficiency in mice through glycolytic metabolic pathway
OBJECTIVE To study the effects of Zigui yichong formula on premature ovarian insufficiency(POI)in mice through glycolysis metabolic pathway.METHODS Eighty SPF C57BL/6N female mice were divided into normal group,model group,Zigui yichong formula group(14.175 g/kg),Zigui yichong formula+2-deoxy-D-arabino-hexose(2-DG)group(Zigui yichong formula 14.175 g/kg+glycolysis inhibitor 2-DG 100 mg/kg),with 20 mice in each group.Except for the normal group,POI model mice were induced by intraperitoneal administration of cyclophosphamide in the other groups.After the model was successfully established,each group was given corresponding drugs.HE staining was employed to observe the pathomorphological changes in ovarian tissue and to count follicles at all developmental stages;radioimmunoassay was conducted to measure the serum levels of estradiol(E2),anti-Müllerian hormone(AMH),and follicle-stimulating hormone(FSH);TUNEL assay was employed to detect apoptosis in ovarian granulosa cells of mice;the activities of hexokinase(HK),pyruvate kinase(PK)and lactate dehydrogenase(LDH)were detected by colorimetry;Western blot and real-time fluorescence quantitative PCR were employed to analyze the protein and mRNA expressions of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),caspase-3,HK2,pyruvate kinase M2(PKM2),and lactate dehydrogenase A(LDHA).RESULTS Compared with model group,the number of primordial follicles,growing follicles,antral follicles and granulosa cells were increased significantly(P＜0.05),and granulosa cells arranged neatly,but the number of atretic follicles and granulosa cells apoptosis were decreased significantly in Zigui yichong formula group(P＜0.05);the serum levels of E2 and AMH,the activities of HK,PK and LDH,protein and mRNA expressions of Bcl-2,HK2,PKM2 and LDHA were increased significantly(P＜0.05);the serum levels of FSH,the protein and mRNA expressions of Bax and caspase-3,Bax/Bcl-2 ratio were decreased significantly(P＜0.05).2-DG could reverse the improvement effects of Zigui yichong formula on the above indexes of POI model mice.CONCLUSIONS Zigui yichong formula may inhibit the apoptosis of ovarian granulosa cells,reduce follicle atresia and improve ovarian reserve function by promoting glycolysis levels in POI model mice.
万方数据
维普
