The Mechanism Of Intracellular Calcium Ionand Integrin Signal Regulating The Adhesion Of Peri-implantation Embryo To Fibronection And The Effects Of Cyclosporine

Objectives: To establish a kind of animal experimental model with peri-implantation embryosof ICR mouse,and to explore the following two points:1.The role of intracellular calcium ion in integrin signaling in regulating the function of adhesion around the implantation embryo and whether cyclosporineaffects this process;2.The calcium ion downstream signaling pathway in the integrin signal regulating the function of adhesion around the implantation embryo and whether cyclosporine affects this process.Methods:1.Embryos were divided into twocyclosporine concentration of 0?mol / L and 1?mol/ L,embryos were sequentially cultured from 1.5dpc to 5.5dpc embryos were co-incubated with FN120 fluorescent microspheres and inactivated FN120 fluorescent microspheres,thenstain the embryo whole cell nuclear with DAPI,and observing the attachment of embryo integrin ?v?3 to fibronectin120 through Confocal laser scanning microscope.2.Embryos were divided into two cyclosporine concentration of 0?mol / L and 1?mol/ L,embryos were sequentially cultured from 1.5dpc to 5.5dpc embryos were co-incubated with calcium ion chelator BAPTA-AM,calmodulin inhibitor W7,protein kinase C inhibitor Calphostin C,W7 and Calphostin C;and the embryos were co-incubated with FN120 fluorescent microspheres,then stain the embryo whole cell nuclear with DAPI,and observing the attachment of embryo integrin ?v?3 to fibronectin120 through Confocal laser scanning microscope.Results:1.The FN120 fluorescence expression of CsA1?M group was higher than CsA0?M group,with significant statistical difference(P<0.01).2.The FN120 fluorescence expression of inactivated FN120 fluorescent microspheres group was lower than CsA group,with significant statistical difference(P<0.01);The FN120 fluorescence expression of inactivated FN120 fluorescent microspheres group was lower than CsA+inactivated FN120 fluorescent microspheres group,with statistical difference(P<0.05).3.The FN120 fluorescence expression of BAPTA-AM group was lower than control group,with statistical difference(P<0.05);The FN120 fluorescence expression of CsA+BAPTA-AM group was lower than CsA group,with significant statistical difference(P<0.01);The FN120 fluorescence expression of BAPTA-AM group was lower than CsA +BAPTA-AM group,with significant statistical difference(P<0.01).4.The FN120 fluorescence expression of W7 group was lower than control group,withsignificant statistical difference(P<0.01);The FN120 fluorescence expression of W7 group was lower than CsA group,with significant statistical difference(P<0.01);The FN120 fluorescence expression of W7 group was lower than CsA +W7 group,with statistical difference(P<0.05).5.The FN120 fluorescence expression of Calphostin C group was lower than control group,with statisticalsignificant difference(P<0.01);The FN120 fluorescence expression of Calphostin C group was lower than CsA group,withsignificant statistical difference(P<0.01);The FN120 fluorescence expression of Calphostin C group was lower than CsA +Calphostin C group,with statistical difference(P<0.05).6.The FN120 fluorescence expression of W7+Calphostin C group was lower than control group,with significantstatistical difference(P<0.01);The FN120 fluorescence expression of W7+Calphostin C group was lower than CsA group,with significant statistical difference(P<0.01);The FN120 fluorescence expression of Calphostin C group was lower than CsA +W7+Calphostin C group,with statistical difference(P<0.01).Conclusion: 3,CsA can further enhance the binding capacity of FN-bound embryos around the bed,this increase may be related to the upregulation of integrin ?v?3 expression.1.After the binding of the peri-implantation embryos to FN,the ability to bind to FN can be further enhanced,making the binding localized in the cell,and in a polar distribution of the embryo.2.The ability to enhance the binding of FN to the peri-implantation embryos may be related to calcium ion and its downstream signaling protein kinase C and calmodulin.3.CsA can further enhance the binding capacity of FN-bound embryos around the peri-implantation,the increasing capacity may be related to the upregulation of integrin ?v?3 expression.