白细胞呼吸暴发形态学改变及银杏叶提取物干预作用

Respiratory Burst of Leukocytes and Intervention by GbE

目的研究白细胞呼吸暴发化学发光活性与细胞形态行为变化特征及其相互关系 ,并探讨GbE对白细胞活化的干预作用。方法体外循环冠脉搭桥病人20例 ,采集转流前升主动脉血样 ,分离白细胞加入FMLP激活剂 ,孵化后置入超弱化学发光系统中 ,计算机描记白细胞呼吸暴发化学发光活性。根据发光曲线分为5个时段 :0=未活化 ,1=活化初期 ,2=活化极期 ,3=活化后期 ,4=失活化。同步用布氏显微镜观察 (放大1000倍 ) ,分析细胞形态行为变化特征,按这5个时段分为 :A、B、C、D和E型5个细胞亚型 ,并作形态、功能相关分析。GbE(含黄酮苷0.545mg/ml)对白细胞呼吸暴发干预试验分为 :对照组 (白细胞 +FMLP)、GbE -1组 (加入GbE5μl)、GbE -2组 (加入GbE10μl)。结果1.FMLP激活白细胞呼吸暴发化学发光计数峰值明显升高 ,显示白细胞氧化暴发释放反应氧类活性的定量表达。2.白细胞活化的极期、后期发光活性与形态活化率呈正相关 (r=0.91 ,0.75) ,也与C型、D型细胞形态改变相一致。但活化初期B型细胞因缺乏脱颗粒释放 ,未能与发光计数平行上升。A型和E型细胞分别反映未活化和失活化状态均相符合。3.GbE对FMLP激活白细胞呼吸暴发化学发光活性有明显浓度依赖性抑制作用。活化极期GbE1组、GbE2组抑制百分率分别为 :23.6 % ,53.35 %(P<0.

Objective. To study the chemiluminescence activity and morphology of leukocytes during respiratory burst and intervention by Ginko biloba extract (GbE).Methods 20 patients undergoing extracorporal circulation (ECC) were observed. Purified leukocyte, blood sample collected from ascending aorta before ECC turning flow. It was stimulated with FMLP and was incubated. Then, described the chemiluminescence activity curve by computer system. The curve can be divided into 0,1,2,3,4 five periods . Meanwhile, at each period, observed the morphology of the leukocytes which can be defined as A,B,C,D and E type. In the GbE groups, additional GbE (GbE-1 group 5ul. GbE-2 group 10ul) was added into the leukocyte samples than the above-mentioned control group.Results The leukocytes released a vast amount of reactive oxygen species (ROS) during respiratory burst; the C type leukocytes' activated rate and the chemiluminescence activity at the curve peak was markedly positive related (r=0.91, P<0.01).Conclusions Bradford variable projective microscopy (BVPM) provides a new rapid diagnostic method for leukocyte activation.There are concentration-dependant inhibition, in chemiluminescence produced by activated leukocyte with FMLP. Additional GBE inhibited percentage showed 23.6%(GBE-1) and 53.35% (GBE-2) compared with control group(P<0.01).