Role of caspase-3 in aggravation of oxidative stress injury in stored red blood cells by CPB: cell experiment
10.3760/cma.j.cn131073.20220919.00208
- VernacularTitle:caspase-3在CPB因素加重储存红细胞氧化应激损伤中的作用：细胞实验
- Author:
Yuling ZHENG
1
;
Hangya LINGHU
;
Daming GOU
Author Information
1. 遵义医科大学附属医院麻醉科，遵义 563003
- Keywords:
Caspase 3;
Erythrocytes;
Extracorporeal circulation;
Oxidative stress
- From:
Chinese Journal of Anesthesiology
2023;43(2):162-165
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the role of caspase-3 in aggravation of the oxidative stress injury in stored red blood cells (sRBCs) by cardiopulmonary bypass (CPB).Methods:Eight patients with type O blood undergoing valve replacement with CPB were selected, blood samples 7 ml were collected through the central venous catheter before CPB, blood samples 20 ml were collected at 2 h after CPB, and the plasma before and after CPB was obtained after centrifugation. Eight samples of sRBCs 14 ml of blood type O stored for 7-14 days were collected and each sample was divided into A, B, C and D groups with 3.5 ml in each group. Normal saline 30 μl was added to group A and group B, 10% dimethyl sulfoxide 30 μl was added to group C, and 3.5 mmol/L Z-DEVD-fmk solution 30 μl was added to group D. The sRBCs were pretreated in a 37 ℃ water bath for 2 h in the four groups. Then group A was incubated with plasma before CPB, group B, C and D were incubated with plasma at 2 h of CPB, and four groups were incubated for 48 h in a thermostatic oscillator at 37 ℃ and 80 rpm. At 2, 24 and 48 h of incubation, the activity of caspase-3 and concentration of ATP were determined by enzyme-linked immunosorbent assay, the concentrations of glutathione (GSH) and free hemoglobin (FHb) were measured by colorimetry, and the exposure rate of cell membrane phosphatidylserine (PS) was detected by flow cytometry.Results:With the prolongation of incubation time, the activity of caspase-3, exposure rate of PS at cell membrane and concentration of FHb were gradually increased, and the concentrations of ATP and GSH were gradually decreased in the four groups ( P<0.05). Compared with group A, the activity of caspase-3 was significantly increased at each time point of incubation in group B and group C, the activity of caspase-3 was increased at 24 and 48 h of incubation in group D, and the concentration of ATP was decreased at 24 and 48 h of incubation, and the concentration of GSH was decreased and the concentration of FHb was increased at each time point of incubation in group B, group C and group D ( P<0.05). Compared with group B and group C, the activity of caspase-3 was significantly decreased, the concentrations of ATP and GSH were increased, and the exposure rate of PS at cell membrane and concentration of FHb were decreased at 24 and 48 h of incubation in group D ( P<0.05). There was no significant difference in the parameters mentioned above between group B and group C ( P>0.05). Conclusions:Caspase-3 is involved in aggravation of oxidative stress injury in sRBCs by CPB.
