离体淋巴细胞经低氧及低氧预处理后免疫指标的变化

Experimental observation of the changes in immune indices of lymphocytes and lymphocytes with hypoxia preconditioning cultured in low O_2 tank

目的 观察急性低氧及低氧预处理后淋巴细胞 [Ca2 + ]i的变化及与淋巴细胞增殖活化的关系。 方法 取正常人外周静脉血 ,用Ficoll Hypaque密度梯度离心法分离淋巴细胞。实验分 3组 :正常对照组、急性低氧组和低氧预处理组。用Fura 2 /AM荧光分光光度法检测 [Ca2 + ]i。用ELISA法检测上清液中IgG和IL 2的分泌量。用MTT法检测淋巴细胞增殖率。 结果 急性低氧后淋巴细胞 [Ca2 + ]i明显下降 ,与对照组比较差异有非常显著性意义 (P <0 0 1) ,同时淋巴细胞IgG和IL 2的分泌量以及淋巴细胞增殖反应也明显下降 ,与各自的对照组相比差异有非常显著性意义(P <0 0 1) ;低氧预处理后再进行急性低氧时 ,淋巴细胞 [Ca2 + ]i、IgG和IL 2的分泌量以及淋巴细胞增殖反应均恢复到与对照组相近的水平 ,且与急性低氧组比较差异有非常显著性意义 (P <0 0 1)。 结论 急性低氧后淋巴细胞免疫功能下降 ,低氧预处理能改善淋巴细胞的免疫功能 ,其中 [Ca2 + ]i的适应性变化发挥了重要作用。

Objective To investigate the effect of hypoxia preconditioning on changes of immune indices and proliferation of lymphocytes cultured in low O 2 tank. Methods Lyphocytes were separated from peripheral venous blood of healthy personnel by Ficoll-Hypaque density gradient centrifugation method. The lymphocyte specimens were divided into 3 groups: normal control group (NC), acutehypoxia group (AH) and hypoxia preconditioning group (HP). The later 2 groups underwent acutehypoxia in a low oxygen tank (1%O 2, 5%CO 2) for 2 h, and HP group experienced intermittent hypoxia(3%O 2, 5%CO 2) for 2 h for 6 times before the acute hypoxia exposure. Lymphocytes [Ca 2+]i was assayed by Fura-2/AM fluorescence method, the levels of IgG and IL-2 in the media were determinedby ELISA method and lymphocyte transformation rate was measured by MTT method. Results In AH group, lymphocyte [Ca 2+]i, lymphocyte IgG and IL-2 secretion levels, as well as lymphocyte transformation rate were lowered significantly (P<0.01) as compared with NC group, while in HP group, all these changes as compared with NC group were non-significant. Conclusions The results suggest that the immune function of lymphocytes was lowered after acute hypoxia exposure, and hypoxia preconditioning can alleviate the change effectively, in which the adaptive change of [Ca 2+]i may play an important role.