化学发光法研究大白兔外周血巨噬细胞中呼吸爆发随年龄的变化

首次建立了检测兔外周血巨噬细胞化学发光的方法，探索了在不同受体刺激剂，例如刀豆蛋白Con A、PMA、N-FMLP等作用下巨噬细胞化学发光信号的变化。实验表明，巨噬细胞化学发光信号随受体刺激剂浓度的提高增大，并在一定条件下与细胞的浓度成线性关系。化学发光试剂鲁米诺的浓度在5×10^-4mol/L、受体刺激剂的浓度在150μg时化学发光强度最大。4种不同的细胞稀释液中，含1% BSA的HBSS用于化学发光的缓冲溶剂效果最佳。应用这一方法对机体年龄增长过程中细胞呼吸爆发程度的变化进行了细致和深入的研究，探索了机体年龄增长与巨噬细胞呼吸爆发间的相互关系。

The Studies on the Respiratory Burst Changes with Age in Rabbit Peritoneal Macrophage with Chemiluminescence Method

The major products of respiratory burst in cells are active oxygen, which include anionic free radical of oxygen, singlet oxygen, hydrogen peroxide and hydroxyl free radical. With the changes of age, the changes of respiratory burst will occur. This phenomenon has a close relationship with the variation of animal body. In this paper, macrophage chemiluminescence(CL) and the function of 3 cell stimulus, Con A, PMA and N-FMLP, have been studied. In the experiment, the intensity of macrophage CL increased with the increasement of stimulus concentration and had linear relationship with the concentration in the range of 100000 to 1000000 cells/mL. The optimal experimental conditions are luminous concentration of 0.1mmol/L, stimulus of 0.00015g/mL for Con A and 0.00020g/mL for PMA, and reaction time of 10 minutes at 37^oC. In 4 types of cells diluents, HBSS-1% BSA is the best buffer solution. The pH range is from 7.2 to 7.4. Using this method to study the relationship between the changes of active oxygen and the age of rabbit, we established a good relation between macrophage CL intensity and the age of rabbit.