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力竭运动应激对淋巴细胞功能及其钾离子通道（Kv1.3）的影响
Researches of Lymphocyte Function and Its Potassium Channels(Kv1.3) Induced by Exercise-Stress
【作者】 傅涛；
【导师】 矫玮；
【作者基本信息】 北京体育大学 ， 运动人体科学， 2008， 博士
【摘要】 目的:通过对免疫细胞上K+通道的研究,探讨运动应激对免疫系统的影响,运动应激触发免疫机能变化的起因。用一种将细胞结构与功能结合起来的系统化模型,做一次深层次认识细胞的尝试。方法:本实验复制了一次力竭运动和两周力竭运动应激模型,取外周血、脾和胸腺组织,采用了淋巴细胞转化实验、分子生物学、药理学、FCM和RVD的方法观察整个过程的结构变化和功能变化。结果:小鼠急性力竭运动应激和两周力竭运动应激后,1.T细胞对ConA诱导的细胞转化功能明显受到抑制。2.RT-PCR和Western blot显示应激组外周血、脾和胸腺的T细胞Kv1.3的mRNA及蛋白的表达水平下降,而应激组外周血和脾的B细胞Kv1.3的mRNA及蛋白的表达与对照组相比没有显著性差异。3.在低渗环境中,应激组小鼠细胞的RVD调节能力明显下降,特别是两周力竭运动应激组。4.细胞周期分析表明:应激组的细胞周期各时期细胞数均发生改变,G0/G1期细胞百分率略有增加,但无显著性差异,S期细胞比率减少,G2/M期细胞的百分数增加,凋亡率增加(P＜0.05)。5.对照组和急性力竭运动应激组CD4/CD8的比值均在正常范围内,两周力竭运动应激组CD4/CD8的比值＞2.0。6.地塞米松和肾上腺素在体外可以明显抑制对照组和应激组小鼠外周血T细胞对ConA诱导的细胞转化功能,并具有量效关系。7.本试验浓度范围内的地塞米松在体外可以明显抑制对照组和急性力竭运动应激组小鼠外周血T细胞的RVD,并具有量效关系,但对两周力竭运动应激组小鼠外周血T细胞RVD均有抑制作用;与地塞米松不同的是无论本实验浓度范围内的肾上腺素均不能抑制对照组和应激组小鼠外周血T细胞的RVD。结论:1.力竭运动应激对T细胞功能影响较大,主要是通过降低T细胞体外活化的反应性来实现的,而对B细胞功能影响很小。2.在运动应激中机体细胞免疫和体液免疫表现出来的差异性,其机制可能与T、B细胞K+通道表达的差异性有关;细胞免疫抑制可能与T细胞上K+通道表达下调有关,钾通道参与了免疫抑制。3.力竭运动应激会影响K+通道介导的RVD功能。4.力竭运动应激组细胞增殖可能受阻于S期;两周力竭运动应激可能会导致细胞免疫紊乱。5.应激激素在体外可抑制对照组和应激组小鼠外周血T细胞的增殖功能,地塞米松可抑制T细胞的RVD功能。
【Abstract】 Object:To explore the interaction of kinesio-stress,lymphocyte and body,We hope establish systematize model that puts together cell function and structure to try to kwon cell.Through researching K+ passage in immunity cell,inquirying kinesio-stress how to influence the immune system,the basic cause of exercise triggering immunity function.Method:In our experiment,we reproduce exhausted sport stress,to appalied lymphocyte transformation experiment,molecular biology, pharmacology and RVD.Result:After experiment,this study arrived at following results: After acute exhausted sport and exhausted sport for two weeks,(1) Spleen T lymphocyte descends to the ConA sensitivity,the function is repressed; (2)RT-PCR and Western blot show the different stress sets of periphery blood,splenic and thymic the mRNA of T lymphocyte of the Kv1.3 and its expression are obviously repressed.Howerve,those of the mRNA of the B lymphocyte Kv1.3 and the expression have no obvious difference with matched control set;(3)In the hypotension environment,exhausted sport sets of RVD regulatory ability descend obviously,especially exhausted sport for two weeks set;(4)The flow cytometry cell cycle analyzes: Compared with matched control set,acute exhausted sport and exhausted sport for two weeks each period cell the number all take place a change, the G0/G1 period cell percentage has increment slightly,but have no statistics difference(P＞0.05),the S period cell significantly decrease, the percentage of the G2/M period cell increase,Apoptosic rate increase(P＜0.05);(5)Exhausted sport sets make the CD4+ cell percent in thespleen T lymphocyte increment(P＞0.05),the CD8+ cell percent also has decreasetrend but hasn’t statistics difference comparison with matched control(P＞0.05).Avalue of CD4+/CD8+ are all in the normal scope,but value of CD4+/CD8+＞2.0 in set of exhausted sport for two weeks.(6)This experiment also confirmed that dexamethasonecan suppress RVD of periphery blood T lymphocyte of normal small mice in vitro and have a quantity effect relation.Dexamethasonecan suppress RVD of periphery blood T lymphocyte of small mice of acute exhausted sport set in vitro and have a quantity effect relation.But different density dexamethasonecan(10-3 mol/L,10-5mol/L,10-7mol/L) can suppress NVD of periphery blood T lymphocyte of small mice of exhausted sport for two weeks set in vitro and haven’ t a quantity effect relation:Conchsion:(1)Two kinds of exhausted sport can influence immune system,through lowering T cell activity,but few influence to the B lymphocyte function:(2)Sport stress regulates the immunity function through the ion passage,which explains the potassium passage participates cell-mediated immunity suppression;(3)The exhausted sport should effect the RVD function of potassium ion passage:(4)The exhausted sport sets of cell proliferation may be got difficulty in S period;(5) The exhausted sport for two weeks should arouse cell immunologic derangement:(6)GC influenced potassium passage of the peripheral blood T cell of mouse of exhausted sport for two weeks,which makes the K+ in the cell outflow decrease,then repress RVD;