Effect of Shoutai Pills on expression of key glycolytic proteins and apoptosis related factors at maternal fetal interface in mouse model of threatened abortion with syndrome of kidney deficiency
This study aims to explore the mechanism of Shoutai Pills in treating threatened abortion.According to the random number table method,ICR female mice were randomized into a normal group,a model group,a dydrogesterone group,and a Shoutai Pills group,with 15 mice in each group.Mice were administrated with normal saline(normal and model groups)or the suspension of Shoutai Pills or dydrogesterone by gavage at 9:00 am every day.At 16:00 every day,mice in the normal group were administrated with an equal volume of distilled water,while those in the model,Shoutai Pills,and dydrogesterone groups were administrated with hydrocortisone solution by gavage for 4 consecutive days.ICR female and male mice were caged in a ratio of 2∶1 during the pre-estrous or estrous period.From the first day of pregnancy,drug administration was continued for 5 consecutive days.On day 6,mice were administrated with mifepristone by gavage to establish the model of kidney deficiency-induced abortion.On day 6 of pregnancy,10 female ICR mice were randomly selected from each group,and the uterus was collected for observation of the pathological changes of trophoblasts at the maternal-fetal interface by hematoxylin-eosin(HE)staining.The protein levels of key enzymes of glycolysis,hexokinase 2(HK2),enolase 1(ENO1),pyruvate kinase M2(PKM2),and lactate dehydrogenase A(LDHA),were determined by Western blot and immunofluorescence.The expression of apoptosis-related proteins including B cell lymphoma-2(Bcl-2),Bcl-2-associated protein X(Bax),and cysteinyl aspartate-specific proteinase-3(caspase-3)was determined by Western blot and real-time PCR.Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling was employed to examine apoptosis.The embryo loss rate of the remaining five female mice was calculated by trypan blue staining method on day 14 of pregnancy.On day 14 of pregnancy,the embryo loss rate of the normal group was 5.00％,which was lower than that(27.78％)in the model group(P＜0.05).Dydrogesterone and Shoutai Pills groups showed reduced embryo loss rates(10.26％and 7.50％,respectively)compared with the model group.On day 6 of pregnancy,compared with the normal group,the model group showed down-regulated expression of HK2,ENO1,PKM2,LDHA,and Bcl-2 and up-regulated expression of Bax and caspase-3(P＜0.05).Compared with the model group,dydrogesterone and Shoutai Pills up-regulated the expression of HK2,ENO1,PKM2,LDHA,and Bcl-2 and down-regulated the expression of Bax and caspase-3(P＜0.05).Compared with that in the normal group,the apoptosis rate in the model group increased(P＜0.05).Compared with the model group,dydrogesterone and Shoutai Pills reduced the apoptosis rate(P＜0.05).In conclusion,Shoutai Pills can reduce the embryo loss rate and protect embryos by promoting aerobic glycolysis at the maternal-fetal interface and inhibiting the apoptosis of trophoblasts in mice.
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