严重烧伤后中性粒细胞对血管内皮细胞形态与功能的影响及参麦注射液对其作用的实验研究

目的 了解严重烧伤后中性粒细胞(PMN)对血管内皮细胞形态与功能的影响及参麦注射液对内皮细胞的保护作用.方法 从健康体检者和烧伤患者(伤后6、24、48 h)静脉血中分离出PMN,与血管内皮细胞株ECV304体外共培养,采用噻唑蓝法检测ECV304细胞活力.放射免疫法检测培养上清液中内皮素1、一氧化氮(NO)的含量,并计算两者的比值(内皮素1/NO).另取伤后24 h血液标本与ECV304共培养,分别加入等渗盐水及0.5、2.0、8.0 g/L参麦注射液各100 μL,观察参麦注射液对上述指标及ECV304形态的影响.结果 与健康体检者比较,在伤后6 h的PMN刺激下ECV304活力明显下降、内皮素1含量明显上升(P＜0.01),伤后24 h的PMN刺激时尤为显著;而伤后6 h在PMN刺激下NO含量明显上升达峰值[(48.9±2.6)μmol/L,P＜0.01],伤后24,48 hPMN刺激ECV304的NO含量与健康体检者比较,无明显差异(P＞0.05);内皮素1/NO的变化趋势同内皮素1.加入不同浓度参麦注射液共培养后,可使内皮细胞活力明显上升、内皮素1含量明显下降、NO含量明显升高,其中中浓度参麦注射液可使体系中内皮细胞形态近似正常.结论 严重烧伤后PMN对血管内皮细胞形态、细胞活力、内皮素1/NO均有不同程度的影响,且以伤后24 h的PMN刺激最明显;参麦注射液可以改善上述情况,对严重烧伤后血管内皮细胞损伤具有保护作用.

Influence of neutrophils from servere burn patients on morphology and function of vascular endothelial cells and the protective effect of intervention with Shenmai concoction

Objective To study the influence of neutrophils from severe burn patient on the morphol-ogy and funetion of vascular endothelial cells(VEC),and the protective effect of Shenmai concoction on VEC. Methods Neutrophils were isolated from the venous blood of healthy volunteers and burn patients[at 6,24,48 post burn hour(PBH)],and they were cocultured with VECs(ECV304)in vitro.The viability of VECs was assessed by MTT.The contents of endothelin(ET)1 and nitric oxide(NO)in supernatant were determined bv radioimmunoassav,the ratio between ET-1 and NO(ET-1/NO) was also calculated.Blood samples from burn patients were collected at 24 PBH to repeat experiment as above,but 100 μL isotonic sa-line or 100μL Shenmai concoction in concentrations of 0.5,2.0,8.0 g/L WaS added to the cultures respec-tivelv.The effect of Shenmai concoction on above indices and morphology of ECV304 were also observed.Results Compared with neutrophils from healthy volunteers.the activity of VECs cocuhured with neutrophils from bum patients at 6 PBH was obviously decreased.the content of ET-1 obviously increased,especially with neutrophils obtained at 24 PBH(P＜0.01).The content of NO in supernatant of VEC co-cultured with neutrophils from burn patients at 6 PBH was increased obviously(48.9±2.6μmoL/L,P＜0.01).There were no obvious difference in the content of NO under the stimulation of neutrophils between that of healthy volunteers and burn patients blood obtained at 24,48 PBH(P＞0.05).With different concentrations of Shenmai.the activity of VECs and the content of NO were obviously increased,while the content of ET-1 was significantly decreased.The morphology of VECs after treatment of neutrophils of 24 PBH and 2.0 g/L Shenmai was 8imilar to normal VECs. Conclusion The morpholo~,activity of VEC,and ET/NO can be affected bv neutrophils after severe burn,especially that of 24 PBH in vitro.Shenmai concoction can amelio-rate above indices.thus might possibly exert protective effects on VECs after severe burn.