运动性贫血时红细胞功能变化以及营养干预对其的影响.doc

运动性贫血时红细胞功能变化以及营养干预对其的影响
摘 要

一、研究目的和意义
运动员的贫血发生率较高。贫血会严重影响运动能力、训练效果、运动后的恢复及免疫等机能状况；有时还成为过度训练的诱因。贫血与体力负荷及营养状况的关系已引起医学界的广泛重视。
本研究的目的是建立运动性贫血的动物模型，并对长期运动训练的大鼠不同时期的红细胞膜变化进行研究，以了解运动训练对红细胞的影响，尤其在大鼠出现运动性贫血时以及潜在性运动性贫血的红细胞膜的变化规律，为准确地反映潜在性贫血和防止运动性贫血的发生和发展提供灵敏监测指标，同时结合血红蛋白、铁代谢参数等指标来评价运动性贫血，以增加对运动性贫血诊断的准确度，为防治运动性贫血的发生和发展提供依据。并对8周运动训练的大鼠红细胞膜变化进行研究，进一步探讨运动性贫血的机理。
二、研究内容和方法
1、大鼠运动性贫血模型的建立
实验中通过10周多级负荷力竭跑台运动建立了运动性贫血模型，并以测定Hb、RBC、HCT来作为评定标准。
2、 8周运动训练及抗运动性贫血剂对大鼠红细胞功能的影响—运动性贫血机制的探讨
本实验在运动性贫血模型基础上和抗运动性贫血剂基础上进行红细胞氧化应激状态、能量代谢功能研究；并利用先进的流式细胞技术和激光共聚焦技术对红细胞的老化进行定量和定性研究；同时利用膜蛋白一维、二维电泳技术观察了红细胞膜蛋白的变化，采用图象分析系统进行红细胞膜蛋白定量分析；通过对上述指标的综合分析，以探讨运动对红细胞损伤以及运动性贫血的机理。
3、 运动性贫血机理和防治措施的研究
本实验对12名贫血运动员及12名正常运动员进行了一系列红细胞指标的测定，并对其进行为期一个月的抗运动性贫血剂的治疗，以探讨运动如何造成红细胞损伤从而导致运动性贫血的机理以及如何进行防治。
三、实验结果
1、大鼠运动性贫血模型的建立
本研究结果显示贫血评定的三个标准指标Hb在10周力竭负荷跑台运动组和对照组之间表现出统计学非常显著性（P＜0.01），而RBC和/或Hct在10周力竭负荷跑台运动组和对照组之间未表现出统计学显著性。此外，由于多级负荷力竭跑台训练持续时间太长（最长时达到一天训练十多小时），而且由于大鼠个体差异较大，从跑台的利用率来说很不经济。所以在正式实验过程中，作者没有采用此种运动性贫血模型，而是采用递增负荷跑台运动造成的运动性贫血模型。
2、 动物实验之8周递增负荷运动训练及抗运动性贫血剂对大鼠红细胞膜功能的影响—运动性贫血机制的探讨
在递增负荷运动所引起的运动性贫血模型上，运动导致红细胞自由基生成增加，脂质过氧化增强，抗氧化酶系统能力降低，Na+ -K+-ATP酶活性降低，红细胞糖酵解和磷酸戊糖旁路两种能量代谢能力均降低，造成对红细胞的损伤。在递增负荷运动所引起的运动性贫血模型上，红细胞老化明显增加，这主要是由于红细胞中的自由基累积增加，抗氧化能力减弱，脂质过氧化增强所致。运动性贫血组的肌动蛋白较对照组明显降低，其原因可能和运动引起的体内自由基的形成和清除的动态平衡紊乱有关，氧自由基可使许多生物大分子如核酸、蛋白质膜多不饱和酸发生损伤，引起超氧化反应，导致膜结构和功能被破坏。同时，本文发现带-6蛋白运动组较对照组明显降低。在递增负荷运动所引起的运动性贫血模型上，抗运动性贫血剂通过降低自由基的生成，并通过不同程度地提高血浆和红细胞的SOD、CAT、GSH-PX水平，改善红细胞糖代谢能力，有效减少红细胞的老化来治疗运动性贫血。
3、 动性贫血及其机理和防治措施的研究
运动性贫血运动员红细胞自由基和脂质过氧化产物增加，抗氧化能力降低，表现为抗氧化酶系统和非酶系统能力均降低。说明运动性贫血运动员红细胞氧化和抗氧化平衡严重失调。使用抗运动贫血剂可明显减少运动后血浆和红细胞MDA的生成，同时提高抗氧化酶系统和非酶系统能力，改善运动员体内血液氧化还原状态。 运动性贫血运动员红细胞糖酵解能力和磷酸旁路代谢能力均降低，ATP和NADPH生成减少，影响机体能量代谢和GSH-PX活性，使用抗运动性贫血剂对磷酸旁路代谢途径有明显的改善作用，但红细胞糖酵解能力则变化不明显。运动性贫血运动员红细胞Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性均降低，红细胞内离子平衡失调，从而影响红细胞膜的渗透性。使用抗运动贫血剂可提高Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性，改善红细胞膜的渗透性和变形性。运动性贫血组的肌动蛋白较对照组明显降低，其原因可能和运动引起的体内自由基的形成和清除的动态平衡紊乱有关，氧自由基可使许多生物大分子如核酸、蛋白质膜多不饱和酸发生损伤，引起超氧化反应，导致膜结构和功能被破坏。运动加快了红细胞老化的过程，运动性贫血组SA较对照组SA有明显降低，PS外翻较对照组PS外翻有明显升高，使用抗运动贫血剂明显延缓红细胞老化的过程，SA有明显升高，PS外翻率明显降低。
四、结论
通过动物和人体实验认为运动导致运动性贫血的机理之一是：（1）递增负荷跑台运动→自由基生成增加，抗氧化能力降低→红细胞氧化应激增加→红细胞膜损伤增加。（2）递增负荷跑台运动后→红细胞无氧酵解的能力减弱，生成的ATP减少，Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性降低，红细胞内离子平衡失调→细胞的渗透性发生改变而影响其变形性，导致红细胞膨胀或脱水，发生溶血→运动性贫血。（3）递增负荷跑台运动后→氧自由基生成增加→红细胞老化增加，溶血率增加→运动性贫血。（4）运动训练后→氧自由基生成增加→攻击红细胞膜蛋白骨架结构→导致膜骨架结构和功能被破坏。
抗运动贫血剂的使用改善了运动员红细胞的氧化应激和能量代谢状态，延缓了红细胞老化过程，对红细胞膜蛋白骨架结构也有一定的改善作用，最终对运动性贫血起到了治疗作用。

关键词 红细胞； 运动性贫血； 抗运动性贫血剂； 自由基； 膜蛋白

Effect of Sports Anemia and Anemia Countermeasures
On Red Blood Cell
(ABSTRACT)
I. Purpose and Significance of the Research
Sports anemia, which often occurs among athletes, can negatively affect athletic performance, training; post-training recovery and the functioning of the athletes’ immune system. Much attention has been given by medical researchers to the relationship between anemia and nutrition.
The purpose of this research is to build an animal model of sports anemia and to monitor red cell membrane changes. The research seeks to determine the effects of training on red cell membrane, especially, when sports anemia results from extended training. This research also seeks to establish some accurate indices for sports anemia. The research evaluates and accurately diagnosis sports anemia using, as references, hemoglobin and ferrum. The research further explores the mechanism of sports anemia.

II. Contents of the Research
Part One: Establishment of a sports anemia model for rats
Two types of training were used to establish the sports anemia model: swimming and treadmill. The three indices used to evaluate this model were: Hb, RBC and HCT.
Part Two: Effect of extended training and anemia countermeasures on red cell of rats
This part of the research investigated the oxidative stress status of blood and the energy metabolism of red blood cells relating to sports anemia and anemia countermeasures. By using the flow cytometer and CLSM (two very modern techniques to study the aging of red blood cells), both qualitative and quantitative analyses were made. Gel electrophoresis was used to determine changes in the membrane protein of red cells, and quantitative analysis of the protein of red cells membrane was achieved through use of an advanced image analysis system.
From the data resulting from use of the above techniques, it was possible to explore how extended training causes damage to red cells and how this, in turn, causes sports anemia.
Part Three: The mechanism of sports anemia, preventive measures, and anemia countermeasures in athletes
In this part of research, two groups of athletes were used: one group of twelve all had sports anemia and the second group of twelve were all healthy. Blood from both groups were studied to establish initial, baseline indices. Following one month of anemia countermeasures, blood was again studied and compared for results. It was determined that extended training caused damage and loss of red cells and also determined that anemia countermeasures could restore red cells in the blood.

III. Experimental Results
Part One: Establishment of a sports anemia model for rats
After ten weeks of exhaustive training on the treadmill, Hb indices in the rats was found to be significantly lower than that of the control group (p<0.01). RBC and HCT were not found to be significantly lower than that of the control group. Also, it takes too long to make this sports anemia model, and can’t make full use of the treadmill because the difference among the rats is too big. So, in the formal experiment, it is suggested to use the progressively more strenuous training on the treadmill to make the sports anemia model.
Part Two: Effect of extended training and anemia countermeasures on the red cell membrane of rats
Based on the sports anemia model of progressively more strenuous training on the treadmill, it was found that such training produced more and more free radicals in the blood; enhanced oxidation and peroxidation; and caused a decrease of serum SOD and Ery-SOD, serum GSH and Ery-GSH, serum CAT and Ery-CAT. This model showed that oxidative stress levels in blood are raised and oxidative injury to red blood cells is induced. This type of exercise was found to impair the energy metabolizing system and to lower the enzyme level of Na+-K+-ATP, which cause damage to the red blood cells. Regarding the senility parameters of red blood cells, the PS extroversion rate was significantly higher than that of the control group and the SA is significantly lower. Anemia countermeasures raised the levels of serum SOD and Ery-SOD, serum GSH and Ery-GSH, serum CAT and Ery-CAT and strengthened the anti-oxidative ability of red cells. Anemia countermeasures, thus effectively regulated oxidation levels in red cells, lowered oxidative stress and reduced senility of red cells due to oxidative stress.
Part Three: The mechanism of sports anemia, anemia preventive measures, and anemia countermeasures in athletes
Compared to the control group the athletes with sports anemia produced more free radicals in the blood; had enhanced oxidation and peroxidation; and their serum SOD and Ery-SOD, serum GSH and Ery-GSH, serum CAT and Ery-CAT levels were lower. With the raising of oxidative stress levels in the blood, oxidative injury to red blood cells was more likely to occur. The athlete’s ATP was lower and their enzyme level of Na+-K+-ATP was also lower, leading to damage of red blood cells. Compared to the senility parameters of the control group, the PS extroversion rate was significantly higher and the SA significantly lower. Following anemia countermeasures, the levels of serum SOD and Ery-SOD, serum GSH and Ery-GSH, serum CAT and Ery-CAT were raised and the anti-oxidative ability of the red blood cells was strengthened. The oxidation level in red cells, thus was effectively regulated by the anemia countermeasures, oxidative stress levels were lowered, and senility of red blood cells, due to oxidative stress, was reduced.

IV. Conclusion
From analysis of the results of the animal model research and of the research on athletes, it can be concluded that one mechanism of sports anemia involves the following exercise-induced factors:
A. Higher production of free radicals and the lowering of oxidation cause oxidative stress levels to rise; thus, leading to damage of red blood cell membrane
B. Impairment of the energy metabolizing system, with lower ATP production and lower Na+-K+-ATP, offsets the ion balance in red cells. This affects the osmotic action in red cell membrane, which leads to swelling or contracting of the cells and, finally, haemolysis and sports anemia.
C. Higher free radical production and higher senility result in the brittleness of red blood cell membrane, causing haemolysis and sports anemia.

Keywords red cell sports anemia anemia countermeasures free radicals
Membrane protein

VII