The Expression Of PKIB And Its Role In Placentas Of Preeclampsia Development

Objective:Preeclampsia(PE)is a common multi-system disease which is unique to human pregnancy,and PE is a serious threat to the health and safety of mothers and infants.It is one of the main causes of maternal and perinatal deaths all over the world.Impaired extravillous trophoblast migration and invasion abilities accompanied by poor spiral vascular remodeling is thought to be the initial reason.In this study,we detected the expression differences of cAMP-dependent protein kinase inhibitor-?(PKIB)in the normal and PE placentas,explored the effect of PKIB on trophoblast function and its involvement in the pathogenesis of PE.We hope to provide new markers for the diagnosis of PE and provide theoretical basis and potential new therapeutic target for the treatment of PE.Methods:(1)We collected the placentas of patients with induced abortion in early pregnancy,PE and normal pregnancy in the 3rd trimester.Immunohistochemical staining and Western Blot were used to calculate the expression level of PKIB in these placentas.(2)The PKIB of HTR8/SVneo trophoblast cell line was specifically knocked down by small interfering RNA(siRNA).The knockdown efficiency was checked by real-time PCR and Western Blot.(3)The migration and invasion abilities of trophoblast were tested by a realtime cellular analyzer(RTCA).(4)Tube formation assay and spheroid sprouting assay were utilized to identify the ability to form vessels of HTR8/SVneo trophoblasts.(5)Western Blot was used to demonstrate the level of phosphorylated Akt of trophoblasts as well as downstream-related genes of Akt signaling pathway in trophoblasts.Results:(1)PKIB was highly expressed in trophoblast of early pregnancy and normal placentas,and the expression level of PKIB in placentas of PE patients was significantly lower than that in normal placentas.(2)Down regulation of PKIB can inhibit the migration,invasion and angiogenesis abilities of HTR8/SVneo cells.(3)Downregulation of PKIB resulted in a decrease in phosphorylated Akt,as well as downstream proteins such as matrix metalloproteinase 2,matrix metalloproteinase 9,and glycogen synthase kinase 3?,which are related to migration and invasion.Conclusion:Our study revealed that the downregulation of PKIB expression may reduce pAkt level and its downstream proteins such as MMP-2,MMP-9 and GSK3 ?,resulted in decreased migration,invasion,and vessel formation abilities by regulating Akt signaling pathway in placental trophoblasts of PE patients.