摘要
目的评价caspase-3在CPB因素加重储存红细胞(sRBC)氧化应激损伤中的作用。方法选取CPB下行瓣膜置换术的O型血患者8例,于CPB前经中心静脉导管取血标本7 ml,于CPB后2 h时再次取血标本20 ml,离心后分别得到CPB前、后血浆备用。取8份O型且储存时间7~14 d的sRBC每份14 ml,每份平均分为4组,每组3.5 ml。A组和B组加入生理盐水30μl,C组加入10%DMSO 30μl,D组加入3.5 mmol/L的Z-DEVD-fmk 30μl;4组于37℃水浴箱中静置2 h。然后A组加入CPB前血浆30μl,B组、C组和D组加入CPB 2 h时血浆各30μl,4组于37℃、80转/min的恒温振荡器中孵育48 h。分别于孵育2、24和48 h时,采用ELISA法检测caspase-3活性和ATP浓度,比色法检测谷胱甘肽(GSH)和游离血红蛋白(FHb)浓度,流式细胞术检测细胞膜磷脂酰丝氨酸(PS)暴露率。结果4组sRBC随孵育时间的延长,caspase-3活性、细胞膜PS暴露率和FHb浓度逐渐升高,ATP和GSH浓度逐渐降低(P<0.05)。与A组比较,B组和C组孵育各时点caspase-3活性升高,D组孵育24和48 h时caspase-3活性升高,B组、C组和D组孵育24和48 h时ATP浓度降低,孵育各时点GSH浓度降低,FHb浓度升高(P<0.05);与B组和C组比较,D组孵育24和48 h时caspase-3活性降低,ATP和GSH浓度升高,细胞膜PS暴露率和FHb浓度降低(P<0.05);B组和C组上述各指标比较差异无统计学意义(P>0.05)。结论caspase-3参与了CPB因素加重sRBC氧化应激损伤的过程。
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 30μl was added to group A and group B,10%dimethyl sulfoxide 30μl was added to group C,and 3.5 mmol/L Z-DEVD-fmk solution 30μl 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
作者
郑玉玲
令狐杭亚
苟大明
Zheng Yuling;Linghu Hangya;Gou Daming(Department of Anesthesiology,Affiliated Hospital of Zunyi Medical University,Zunyi 563003,China;Department of Anesthesiology,Kweichou Moutai Hospital,Zunyi 564512,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2023年第2期162-165,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(81860083)。
