摘要
目的了解琥珀酸对人外周血中性粒细胞（PMN）凋亡的影响，探讨其致病机制。方法体外孵育PMN，将PMN浓度调至5×10^6个／ml。在细胞中加入琥珀酸刺激，根据所加琥珀酸浓度分为5、10、20、30mmol／L琥珀酸组；对照组加入常规培养液。取各组细胞培养上清液，检测其活性氧含量。5、20mmol／L琥珀酸组细胞于处理前及处理后2、4、6、8、10h分别取细胞悬液1ml，行半胱氨酸天冬氨酸蛋白酶3（caspase-3）活性及细胞凋亡率检测。结果5、10、20、30mmol／L琥珀酸组活性氧含量（0．437±0．056、0．432±0．024、0．395±0．049、0．386±0．010）均低于对照组（0．505±0．028，P〈0．05）。随着孵育时间延长，各组细胞caspase-3活性均逐渐增加，但与对照组比较，5mmol／L琥珀酸组caspase-3活性降低，20mmol／L琥珀酸组则升高（P〈0．05）。对照组细胞处理前凋亡率为（6．1±1．1）％，处理后2h为（13．2±2．0）％，10h达（27．7±3．7）％；5mmol／L琥珀酸组细胞凋亡率除处理后4h外其余时相点均低于对照组（P〈0．05）；20mmol／L琥珀酸组细胞处理后4～10h凋亡率均高于对照组（P〈0．05）。结论低浓度琥珀酸抑制PMN凋亡，而高浓度琥珀酸可促进PMN凋亡。细菌感染时，通过代谢产物琥珀酸抑制PMN免疫功能。
Objective To investigate the influence of succinic acid on the apoptosis of polymorphonuclear neutrophil （PMN） in human peripheral blood, and to explore its role in infection. Methods PMNs were incubated in vitro, and its concentration was adjusted to 5 × 10^6/mL. Then the ceils were divided into normal control group and 5,10,20,30 mmol/L succinic acid groups according to different concentrations of succinic acid added into the medium. The supernatant of the cultures in each groups were collected to deter- mine the superoxide content. 1 mL cell suspension was collected from 5,20 mmol/L succinic acid groups be- fore treatment and at 2,4,6,8,10 post-treatment hours（PTH） for the determination of caspase-3 activity and the apoptosis rate. Results The content of superoxide in 5,10,20,30 mmol/L succinic acid groups （0. 437 ± 0. 056,0. 432 ± 0. 024 ,0. 395± 0. 049 ,0. 386 ± 0. 010） was significantly lower than that in control group（ 0. 505 ± 0. 028, P 〈 0.05 ）. The caspase-3 activity in each group increased along with the incubation time ,but was in lower concentration in 5 mmol/L succinic acid group and in higher concentration in 20 mmol/L succinic acid group when compared with that in control group（ P 〈 0.05）. The apoptosis rate of PMN in control group was （ 6.1 ± 1.1 ） % before incubation, and it reached （13.2 ± 2.0）% at 2 PTH,and （27.7 ± 3.7 ） % at 10 PTH. The apoptosis rate of PMN in 5 mmol/L succinic acid group was lower than that in control group except that at 4 PTH （ P 〈 0.05）. On the other hand, the apoptosis rate in 20 mmol/L suecinic acid group（ during 4-10 PTH） were obviously higher at each time points compared with the control group （ P 〈 0.05）. Conclusion Low concentration of succinic acid can suppress the apoptosis of PMN, while high concentration of succinic acid has an opposite effect. It is known that bacteria can produce succinic acid.
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2007年第6期417-419,共3页
Chinese Journal of Burns
基金
国家自然科学基金(30471785
30571920)
关键词
琥珀酸
中性白细胞
细胞凋亡
Succinic acid
Polymorphonuclear neutrophil
Apoptosis
