Mechanism Of Kisspeptin Regulating Endometrial Decidualization In Recurrent Spontaneous Abortion By ERK1/2 Signaling Molecule

Objective:In the present study,Kisspeptin was involved in the regulation of endometrial decidualization,and the role and potential mechanism of Kisspeptin in the pathogenesis of recurrent spontaneous abortion were investigated.Methods:1.Clinical research:immunohistochemistry and Western-blot methods were used to detect the expression levels of proteins in 32 patients with unexplained recurrent spontaneous abortion(URSA)and 34 women with normal early pregnancy,including kisspeptin?p-ERK?ERK in the villi and decidua and IGFBP-1?dPRL in the decidua.2.Animal experiments:(1)establish recurrent spontaneous abortion model CBA/JxDBA/2 and normal pregnancy model CBA/J×BALB/c,and the number of aborted embryos was counted on the 9th and 14th day of pregnancy,the decidual tissue and placenta of pregnant rats were taken out for kisspeptin,p-ERK,ERK,IGFBP-1,dPRL immunohistochemical staining and Western-blot detection on the 9th day of pregnancy.(2)the pregnant mice in the above recurrent spontaneous abortion model were randomly divided into experimental group 1,experimental group 2 and control group.From the day of conception,the kisspeptin peptide 100nmol/kg(2.6?g/only)was injected into the tail vein every other day for the experimental group 1(KP),the injection of the kisspeptin receptor blocker 100nmol/kg(2.6?g/only)for the experimental group 2(KB),and the control group(NS)were injected with the same amount of saline.The number of aborted embryos was counted on the 9th and 14th day of pregnancy,and the differences between the groups were compared.the decidual tissue and placenta of pregnant rats were taken out for kisspeptin,p-ERK,ERK,IGFBP-1,dPRL immunohistochemical staining and Western-blot detection on the 9th day of pregnancy.To observe the effect of kisspeptin addition on the expression of p-ERK?ERK?IGFBP-1?dPRL,and the effect of kisspeptin receptor blocking on the expression of p-ERK?ERK?IGFBP-1?dPRL.Results:1.The expression of kisspeptin and p-ERK in the villi of RSA patients was significantly lower than that in normal pregnancy(P<0.05).The expression of kisspeptin,p-ERK,IGFBP-1 and dPRL in decidua was significantly lower than that in normal pregnancy(P<0.05),while the expression of ERK was not significantly different in villi and decidua(P>0.05).2.Compared with the normal pregnancy model CBA/J×BALB/c,the embryo absorption rates of the recurrent spontaneous abortion model CBA/J×DBA/2 pregnancy on the 9d and 14d were significantly higher than that of the normal pregnancy model(29.17%± 2.35%vs 9.13%± 1.90%/9d,30.00%± 2.65%vs 8.20%± 1.64%/14d)(P<0.05).Meanwhile,the expressions of kisspeptin?P-ERK?IGFBP-1?dPRL in the matemal-fetal interface of CBA/J×DBA/2 mice were significantly lower than that of CBA/JxBALB/c mice(P<0.05),while the expressions of ERK were not significantly different at the maternal-fetal interface(P>0.05).3.Exogenous kisspeptin supplementation decreased the embryo absorption rate of CBA/JxDBA/2 mice compared with the normal saline control group(7.30%± 1.18%vs 28.70%± 1.95%/9d,10.41%± 2.07%vs 27.29%± 3.06%/14d)(p<0.01).The enhanced expressions of p-ERK?IGFBP-1?dPRL at maternal-fetal interface were detected(P<0.05).There was no statistically significant difference between the changes of CBA/JxDBA/2 mouse embryo absorption rate after blocking kisspeptin receptor(28.03%±2.53%vs 28.70%±1.95%/9d,29.27%± 3.01%vs 27.29%± 3.06%/14d)(p>0.05).However,the expressions of p-ERK?IGFBP-1?dPRL at the matemal-fetal interface were decreased(P<0.05).Conclusions:1.The decreased expression of kisspeptin in the matemal-fetal interface of RSA patients suggested that the abnormal expression of kisspeptin was associated with the onset of abortion in RSA patients.2.In patients with recurrent spontaneous abortion,the expression of decidua kisspeptin was decreased,and expression of p-ERK and the decidualization markers(IGFBP-1 and dPRL)were also low.3.It was confirmed in animal experiments that kisspeptin can improve decidualization indexes,up-regulate the expression of p-ERK and reduce the abortion rate.We speculated that kisspeptin can regulate uterine decidualization,this regulation may be achieved by ERK1/2 signaling molecules.Low expression of kisspeptin may lead to miscarriage caused by poor decidualization.