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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 observe the changes of [Ca2 +] i in lymphocytes and their relationship with lymphocyte proliferation and activation after acute hypoxia and hypoxia preconditioning. Methods Peripheral venous blood was collected from healthy volunteers and lymphocytes were isolated by Ficoll Hypaque density gradient centrifugation. The experiment was divided into 3 groups: normal control group, acute hypoxia group and hypoxic preconditioning group. [Ca2 +] i was measured by Fura 2 / AM spectrofluorimetry. The amount of secreted IgG and IL 2 in the supernatant was measured by ELISA. Lymphocyte proliferation rate was detected by MTT assay. Results The level of [Ca2 +] i in lymphocytes was significantly decreased after acute hypoxia compared with the control group (P <0.01). At the same time, the secretion of lymphocytes IgG and IL-2 and the lymphocyte proliferation reaction were also significant (P <0.01). After hypoxic preconditioning, the levels of [Ca2 +] i, IgG and IL2 secretion in acute hypoxia were significantly lower than those in control group (P <0.01), and the proliferation of lymphocytes returned to the same level as the control group, and the difference was significant compared with the acute hypoxia group (P <0.01). Conclusion The immune function of lymphocytes is decreased after acute hypoxia. Hypoxic preconditioning can improve the immune function of lymphocytes, and the adaptive changes of [Ca2 +] i play an important role.