Kisspeptin Cooperate With Progesterone To Regulate Endometrial Decidualization Via Notch1 Signal Path Way

Purpose:This study is aimed to detect the difference of target genes of Notch1 signal pathway in decidual tissue between unexplained recurrent spontaneous abortion(URSA)patients and normal early pregnant women in regulating endometrial decidualization,then verify protein expression levels of some target genes.Then we study the relationship between kisspeptin and target genes,to test the hepothesis that kisspeptin cooperating with progesterone mediates endometrial decidualization,via Notch1 signal path way.Method:1.During March 2019 to December 2019,we collect decidua issue of 11 women with URSA and 10 normal early pregnant women who were treated in the reproductive clinic of the Second Affiliated Hospital of Soochow University,use mRNA array to detect the difference gene expression of Notch 1 signal path way between two groups.2.Based on the results of the mRNA array,we selected Notch1,Jag2,and Hes5.Then we collect decidua issue of 19 women with URSA and 12 normal early pregnant women who were treated in the reproductive clinic of the Second Affiliated Hospital of Soochow University during March 2019 to December 2019,and detected kesspeptin,Notch1,Jag2,and Hes5 protein expression by immunohistochemistry and Western-blot.3.Cell experiments:(1)Human endometrial stromal cells(hESCs)were cultured in vitro.We used E2?MAP and 8-Br-cAMP to induce hESCs decidualization,and investigated gene and protein expression levels of Notch1 and Jag2,Hes5 over time by RT-PCR and western blot.(2)Cell grouping:hESCs were divided into three groups:Control group:normal decidualization;Kisspeptin group:kisspeptin was added;siRNA group:Kiss-siRNA transfection in hESCs.Then we compared gene and protein expression of Notch1 and its related molecules Jag2,Hes5 levels between the three groups by RT-PCR and western blot.Results:1.Notch 1 pathway-related target genes are down-regulated in decidual tissues of URSA when comparing to normal women.The expression of NOTCH 1,HES5,JAG2,LOR,SH2D1A,FZD2,DLL3,LFNG,PAX5,POFUT1,IL178 and so on decreased in URSA more than 3 times over normal women.2.Protein expression of kisspeptin,Notch1,Jag2 and Hes5 in URSA patients are significantly lower than normal pregnancy(P<0.05).3.Stromal cells were observed by light microscope show typical changes of cell enlargement and roundness and nucleus enlargement in the process of decidualization.The genes and proteins expression of Notch 1 and its related molecules jag2 and Hes5 increased gradually(P<0.05)from 24 h to 96 h.The protein expression of Notch1 in kisspeptin group increased.The protein expression of Notchl in siRNA group was lower than control group.The expression of Notch1,jag2 and Hes5 genes was up-regulated in kisspeptin group and down regulated in siRNA group(P<0.05).Conclusions:1.Notch1 family members are down regulated in decidua of URSA patients,suggesting that abnormal expression of Notchl signal pathway is related to the etiology of URSA.2.The expression of kisspeptin,Notch1,jag2 and Hes5 protein in decidua of URSA patients decreased.Kisspeptin promotes the expression of Notch1 and its related molecules Jag2 and Hes5.Kisspeptin may cooperate with progesterone via Notch1 signal path way to regulate endometrial decidualization.