The Research Of Caspase-3 Inhibitor(Z-DEVD-fmk) On Protective Effect Of SRBC Incubated By Plasma After CPB

Objective: To observe the protective effects of caspase-3 inhibitor(Z-DEVD-fmk)on sRBC incubated by plasma after CPB Methods:1.Preparation of plasma at different time points of CBP: Eight patients undergoing valve replacement with CPB were selected(n=8),7 mL blood was taken through central venous catheter before CPB,and 20 mL blood was taken again after CPB 2 hours.After centrifugation,the plasma before and after CPB was obtained.2.Grouping and pretreatment of sRBC: 14 mL sRBC stored for 7-14 days were equally divided into group A,B,C and D(3.5 mL in each group).30μL NS was added to group A and group B;30μL of 10% DMSO diluent was added to group C;30μL of 3.5m M Z-DEVD-fmk solution was added to group D.All groups were incubated at 37℃ for 2 hours.3.Plasma before and after CBP incubated with sRBC: The sRBC of group A was incubated with plasma before CPB with the equal volume;sRBC of group B,C and D were incubated with the equal volume of plasma after CPB 2 hours,and four groups were incubated for 48 hours.4.The sRBC was took out and detected the activity of caspase-3,the content of ATP and GSH content as well as the PS exposure rate on erythrocyte membrane and the concentration of FHb in incubation solution at 2,24 and 48 hours respectively in all groups.Results:1.Changes of the caspase-3 activity of sRBC in four groups at different incubation time.The caspase-3 activity of sRBC was significantly lower in group A than that in group B and group C(P < 0.01),but there was no statistical difference not only between group A and group D(P > 0.05),but also among group B,C,D after incubation for 2 hours.It was significantly lower in group A than that in group B,C and D(P < 0.01),but there was no statistical difference between group B and group C(P > 0.05),while it was significantly lower in group D than that in group C(P < 0.01)after incubation for 24 and 48 hours.2.Changes of the ATP content of sRBC in four groups at different incubation time.There was no significant difference in the ATP content of sRBC among all groups(P > 0.05)after incubation for 2 hours.It was significantly higher in group A than that in group B,C and D(P < 0.01),but there was no statistical difference between group B and group C(P >0.05),while it was significantly higher in group D than that in group C(P < 0.01)after incubation for 24 and 48 hours.3.Changes of the GSH content of sRBC in four groups at different incubation time.The GSH content of sRBC was significantly higher in group A than that in group B,C and D(P < 0.01),but there was no statistical difference among group B,C,D(P > 0.05)after incubation for 2 hours.It was significantly higher in group A than that in group B,C and D(P < 0.01),but there was no statistical difference between group B and group C(P > 0.05),while it was significantly higher in group D than that in group C(P < 0.01)after incubation for 24 and 48 hours.4.Changes of the PS exposure rate on sRBC membrane in four groups at different incubation time.The exposure rate of PS was significantly lower in group A than that in group B,C and D(P < 0.01),but there was no statistical difference among group B,C,D(P >0.05)after incubation for 2 hours.It was significantly lower in group A than that in group B,C and D(P < 0.05),but there was no statistical difference between group B and group C(P >0.05),while it was significantly lower in group D than that in group C(P < 0.01)after incubation for 24 and 48 hours.5.Changes of the FHb concentration of the incubation solution in four groups at different incubation time.The FHb concentration of the incubation solution in group A was significantly lower than that in group B,C and D(P < 0.01),but there was no statistical difference among group B,C,D(P > 0.05)after incubation for 2 hours.It was significantly lower in group A than that in group B,C and D(P < 0.01),but there was no statistical difference between group B and group C(P > 0.05),while it was significantly lower in group D than that in group C(P < 0.05)after incubation for 24 and 48 hours.Conclusion:1.The plasma of patients after CPB has caused damage to sRBC.2.Z-DEVD-fmk has protective effects on sRBC incubated by plasma after CPB.