The Mechanism Of Bushen Antai Granule Regulating ERK1/2 On Deciduating RSA Mice By Kisspeptin

1 ObjectiveTo observe the effects of Bushun Antuan Granule on embryo loss rate,morphological changes of decidua tissue,protein distribution and expression levels of Kisspeptin,ERK1/2,P-ERK,dPRL,IGFBP-1 in decidua tissue of recurrent abortion mice,and then discuss the mechanism of Bushun Antuan granule to protect fetus from the perspective of endometrial decidua.It provides a new theoretical basis for the treatment of RSA by Bushen Antai granule in clinic.2 MethodsIn this study,30 Clark classical RSA model mice were randomly divided into model group,western medicine group,Chinese medicine low-dose,medium-dose and high-dose groups,with 6 mice in each group.There were 6 BALB/c ×CBA/J normal pregnancy mice.Normal group and model group were given equal volume of distilled water.The western medicine group was given progesterone capsules intragastric administration;The Chinese medicine group was given different doses of Bushen Antai infusion.The drug was continued for 8 days,and samples were collected 24 h after the last administration.The decidual tissues of mice in each group were separated and collected,and the loss of embryos in each group was recorded.The protein distribution and expression levels of Kisspeptin,ERK1/2,P-ERK,dPRL and IGFBP-1 in decidua tissue of pregnant mice were detected by immunofluorescence and Western bloting.3 Results3.1 Embryo loss rate:The embryo loss rate in model group was significantly higher than that in normal group(P<0.01).Compared with model group,the rate of embryo loss in western medicine group and Bushen Antai granule low,medium and high dose groups was significantly decreased,and the difference was statistically significant(P<0.01).The embryo loss rate of Bushen Antuo granule high dose group was lower than that of medium and low dose groups(P < 0.01);Compared with western medicine group,there was no significant difference(P>0.05).3.2 HE staining of decidua tissue:Compared with the model group,the decidual tissue morphology,cell distribution and nucleus morphology of pregnant mice in the drug treatment group were significantly improved,the decidual blood vessel supply was increased,and the vascular stasis was significantly reduced.The number of cells in decidual tissue was significantly improved in the western medicine group and the Chinese medicine group,and the progressive staining of cytoplasm and cytoplasm was normal and abundant,and the morphology of nuclei tended to be normal.The shape and number of blood vessels became normal.3.3 Detection of relevant indicators in decidua tissue by immunofluorescence method:The m RNA expressions of Kisspeptin,dPRL,IGFBP-1 and ERK1/2 in model group were lower than those in normal group(P < 0.01).Compared with model group,m RNA expression levels of Kisspeptin,dPRL,IGFBP-1 and ERK1/2 in western medicine group and Bushenantuan Granule high,medium and low dose groups were significantly increased(P < 0.01).Bushen Antai granule high dose group3.4 Western Blot was used to detect the expression of related proteins in decidua tissue:Kisspeptin,P-ERK,dPRL,IGFBP-1 proteins in normal group were higher expressed than those in model group(P < 0.01).Compared with model group,protein expressions of Kisspeptin,P-ERK,dPRL,IGFBP-1 in western medicine group,Bushenantuan Granule low,medium and high dose groups were significantly increased(P < 0.01);The protein expressions of Kisspeptin,P-ERK,dPRL,IGFBP-1 in Bushenantai granule high dose group were significantly increased compared with those in medium and low dose groups(P < 0.01),but there was no statistical significance compared with western medicine group(P >0.05).There was statistical difference between Bushen Antuan Granule high-dose group and normal pregnancy group(P <0.01).There were no significant differences in the expression of ERK1/2 protein among all groups(P>0.05).4 ConclusionBushenantuan granule can reduce the rate of embryo loss,improve the morphology of decidual tissue of RSA mice,and up-regulate the protein expression of Kisspeptin,P-ERK,dPRL,IGFBP-1 in the decidual tissue.The mechanism of ERK1/2 signaling molecule regulation with kisspepitn may be involved in endometrial decidualization process.