长期自主转轮跑运动对小鼠脾淋巴细胞钙稳态,IL-2分泌及钙调基因表达的影响

Effects of Long-duration Willing Running-on-wheels on Intracellular Ca^(2+) Homeostasis,IL-2 Secretion and Ca^(2+)-regulating Genes Expression of Lymphocytes in Mouse Spleens

目的:通过构建动物模型,研究运动对淋巴细胞钙稳态、IL-2分泌及钙调基因表达影响,以探讨长期适量强度运动调控免疫机能的分子机制。方法:相互匹配雄性小白鼠随机被分成控制组和实验组,实验组执行长期自主转轮跑运动6个月,淋巴细胞分离自小鼠脾脏,胞内钙离子浓度通过荧光稳态/瞬态分析系统测量荧光探针负载细胞悬液而确定,ELISA法检测Con A体外刺激对淋巴细胞分泌IL-2水平的影响,实时荧光定量PCR法检测细胞相关钙调基因表达。结果:不管在含钙缓冲液还是在不含钙PBS溶液中,实验组OKT-3诱导淋巴细胞内钙离子浓度变化幅度显著高于控制组(P<0.05,n=5);实验组Con A体外刺激诱导培养细胞IL-2分泌水平显著高于控制组(P<0.05,n=6);实验组淋巴细胞内钙调节基因STIM1和ORAI1核酸表达水平显著低于控制组(P<0.05,n=5)。结论:长期自主转轮跑运动可促进小鼠脾淋巴细胞钙离子信号转导敏感性,提高体外刺激诱发淋巴细胞分泌IL-2水平,推测胞内钙稳态调控基因表达下调可能是一种负反馈机制,以防止运动诱发的胞内钙稳态失衡,当然这仍需进一步的实验深入研究运动的调控机理。

By using animal models, the purpose of this study is to examine the effect of long-duration willing wheel-running on intracellular Ca2＋ homeostasis, expression of Ca2＋-regulating genes and mitogen-induced secretion of IL-2 in lymphocytes of mouse spleens and to explore the molecular mechanism that how the long-duration and regular physical activities regulates the immune function. The male mice were divided to the two groups, the experimental group and the control group at random. The mice in the experimental group performed long-duration willing wheel-running for 6 months. Lymphocytes were separated from mouse spleens. Intracellular Ca2＋ was decided by using a spectrometer to measure fluorescent probe-loaded lymphocytes suspension. The effect of Con A as a stimulant in vitro culture on the secretion of IL-2 in lymphocyte was determined by using ELISA. The expressions of Ca2＋-regulating genes were decided by real-time PCR analysis. Either in Ca2＋-containing buffer or in Ca2＋-free PBS solution, OKT3-induced change of intracellular Ca2＋ concentration was significantly higher in the experimental group than in the control group （ P〈0.05, n=5 ）. The secretion of IL-2 is higher in the experimental group than in the control group by using Con A as a stimulant in vitro culture （P〈0.05, n=6 ）. However, the expression of mRNA, such as, Ca2＋ -regulating genes, STIM1 , ORAI1 , were down-regulated, significantly in the experimental group （ P〈0.05 or 0.01 , n=5 ）. This research suggested that long-duration willing running on wheels promotes the sensibility of intracellular Ca2＋ signals, and IL-2 secretion of in lymphocytes of mouse spleens. And we speculate that down-regulation of the Ca2＋ regulating genes expression might serve as a reverse feedback mechanism to protect ceils from the unbalanced intracellular Ca2＋ homeostasis. Certainly it needs further experiments to investigate the mechanism that physical activities regulate intracellular Ca2＋ homeostasis of lymphocytes.