一次性大负荷运动对淋巴细胞钙瞬态、通道基因表达和刺激剂诱导细胞增殖能力的影响

发布时间：2018-12-20 18:21

【摘要】：目的:淋巴细胞增殖反应通常被用作细胞免疫功能的检测指标,运动性免疫抑制是运动医学研究的一个热点,然而其机制至今不甚明了。通过构建一次性大负荷运动模型,研究急性运动对淋巴细胞钙离子瞬态、钙通道基因表达和增殖功能的影响,从而探讨运动性免疫抑制的分子机制。方法:相互匹配的雄性小白鼠随机被分成控制组和运动组(分为运动后即刻处死组和运动后3 h处死组);运动组执行80%最大摄氧量强度跑台运动至力竭;淋巴细胞分离自小鼠脾脏;淋巴细胞内钙离子浓度通过荧光稳态/瞬态分析系统测量fura-2(AM)负载细胞悬液而确定;结合流式细胞术与羧基荧光素琥珀酰亚胺酯(CFSE)负载法评估细胞增殖反应;实时荧光定量PCR法检测淋巴细胞相关通道基因表达。结果:在运动后即刻处死组,植物血凝素(PHA)诱导淋巴细胞内游离钙离子浓度变化幅度和钙离子释放率显著低于控制组(P0.05);在运动后3 h处死组,毒胡萝卜素(TG)诱导淋巴细胞钙离子释放率显著低于控制组(P0.05),且淋巴细胞内钙通道基因RYR核酸表达水平显著高于控制组(P0.05),但细胞膜钙通道基因STIM表达在两组之间没有显著性差异(P0.05),刺激剂体外刺激诱导细胞增殖水平显著低于控制组(P0.05)。结论:急性大负荷运动可能通过下调淋巴细胞内质网网膜钙通道基因表达,抑制胞内钙离子释放以及胞内Ca~(2+)信号转导过程,相应降低刺激剂诱导淋巴细胞增殖能力,即:淋巴细胞免疫功能削弱。这可能是急性运动引起机体免疫抑制的一个重要分子机制。
[Abstract]:Objective: lymphocyte proliferation is usually used as an indicator for the detection of cellular immune function. Exercise immunosuppression is a hot topic in sports medicine, but its mechanism is still unclear. The effects of acute exercise on the transient calcium ion expression, calcium channel gene expression and proliferation of lymphocytes were studied in order to explore the molecular mechanism of exercise induced immunosuppression. Methods: the matched male mice were randomly divided into control group and exercise group (divided into two groups: immediately after exercise and 3 hours after exercise). The concentration of calcium in lymphocytes was determined by fluorescence steady-state / transient analysis system to measure the suspension of fura-2 (AM) loaded cells. Flow cytometry combined with carboxyfluorescein succinimide (CFSE) loading assay was used to evaluate cell proliferation and real-time fluorescence quantitative PCR was used to detect the expression of lymphocyte associated channel genes. Results: immediately after exercise, the changes of free calcium concentration and calcium release rate in lymphocytes induced by phytohemagglutinin (PHA) were significantly lower than those in control group (P0.05). At 3 h after exercise, the calcium release rate of lymphocytes induced by carotene (TG) was significantly lower than that of control group (P0.05), and the expression level of calcium channel gene RYR nucleic acid in lymphocytes was significantly higher than that in control group (P0.05). However, there was no significant difference in the expression of calcium channel gene STIM between the two groups (P0.05), and the level of cell proliferation induced by stimulator in vitro was significantly lower than that in control group (P0.05). Conclusion: acute heavy exercise may inhibit intracellular calcium release and Ca~ _ (2) signal transduction by down-regulating the expression of calcium channel gene in endoplasmic reticulum (ER) of lymphocytes and decrease the ability of stimulating agents to induce lymphocyte proliferation. Namely: lymphocyte immune function weakens. This may be an important molecular mechanism of immune suppression induced by acute exercise.
【作者单位】：中国地质大学(武汉);湘南学院;
【基金】：中央高校基本科研业务费专项资金项目(CUG150607)
【分类号】：G804.7

【共引文献】