Effects of training on spleen and peritoneal exudate reactive oxygen species and lymphocyte proliferation by splenocytes at rest and after an acute bout of exercise

The purpose of this study was to analyse the effect of regular exercise on spleen and peritoneal exudate reactive oxygen species (ROS) and lymphocyte proliferation by splenocytes. Twenty-four female BALB/c mice were randomly divided into trained (n = 12) and untrained (n = 12) groups. These two groups were further divided into mice that were studied at rest (trained/rest, n = 5; untrained/rest, n = 6) and immediately after a 2 h acute bout of exercise (trained/exercise, n = 6; untrained/exercise, n = 6). The animals were bred in the animal facility of the Yonsei University College of Medicine, where they were housed in a temperature- (22 - 24 degrees Celsius) and humidity- (50 - 60%) controlled environment, with a 12 h photoperiod, and provided with food and water ad libitum. The trained mice underwent 10 weeks of endurance swimming training (5 days per week) in water at 26 - 29 degrees Celsius for 60 min. Changes in body mass, proliferative activity and the production of reactive oxygen species from spleen lymphocytes and peritoneal exudate cells were determined. The splenic lymphocytes of the trained mice had much greater proliferative activity than those of the untrained mice (P < 0.05). Trained mice had lower ROS production in splenic lymphocytes and peritoneal exudate cells than untrained mice. In both groups, there was substantial inhibition of proliferative activity stimulated with medium, concanavalin A and lipopolysaccharide following the acute bout of exercise. This may have been caused by excessive ROS production following the acute exercise session.     

长期运动训练对外周血淋巴细胞DNA损伤和凋亡的影响及机制研究

训练组(男子足球运动员,n=16)和对照组(男子大学生,n=16)在功率自行车上完成一次递增负荷力竭运动。运动前、运动后以及运动后24 h采集静脉血,测定外周血淋巴细胞计数、淋巴细胞凋亡率、羟自由基、8-羟基脱氧鸟嘌呤(8-OHdG)和8-氧鸟嘌呤DNA糖基化酶(OGG1)。结果发现,运动后即刻,训练组和对照组外周血淋巴细胞计数(P<0.01;P<0.05)、淋巴细胞凋亡率(P<0.05;P<0.01)、羟自由基(P<0.01;P<0.01)、8-OHdG(P<0.01;P<0.01)和OGG1(P<0.01;P<0.01)均高于运动前水平,而训练组OGG1升高幅度高于对照组(P<0.01),其他各指标升高幅度则均明显低于对照组(均为P<0.01)。运动后24h,对照组外周血淋巴细胞计数低于运动前(P<0.01),淋巴细胞凋亡率高于运动前(P<0.01),而训练组则恢复至运动前;两组8-OHdG(P<0.05;P<0.01)均高于运动前,但升高幅度明显低于对照组(P<0.01);OGG1在对照组恢复至运动前,而训练组仍高于运动前水平(P<0.05)。因此,长期运动训练通过降低氧化应激水平、提高DNA修复能力、减轻DNA氧化损伤,抑制运动性淋巴细胞凋亡。     

Mechanisms of beneficial effects of exercise training on non-alcoholic fatty liver disease (NAFLD): Roles of oxidative stress and inflammation

Abstract

Non-alcoholic fatty liver disease (NAFLD) is a common chronic liver disorder which is associated with accumulation of fats in the liver. It causes a wide variety of pathological effects such as non-alcoholic steatohepatitis (NASH) and cirrhosis, insulin resistance, obesity, hypertension, dyslipidaemia, diabetes and cardiovascular disease. The molecular mechanisms that cause the initiation and progression of NAFLD are not fully understood. Oxidative stress (OS) induced by reactive oxygen species (ROS) and inflammation are likely a significant mechanism which can lead to hepatic cell death and tissue injury. Mitochondrial abnormalities, down-regulation of several antioxidant enzymes, glutathione (GSH) depletion and decreased activity of GSH-dependent antioxidants, accumulation of leukocytes and hepatic inflammation are the major sources of ROS overproduction in NAFLD. Excessive production of ROS suppresses the capacity of other antioxidant defence systems in NAFLD and causes further oxidative damage. Regular exercise can be considered as an effective strategy for treatment of NAFLD. It improves NAFLD by reducing intrahepatic fat content, increasing β-oxidation of fatty acids, inducing hepato-protective autophagy, overexpressing peroxisome proliferator-activated receptor- γ (PPAR-γ), as well as attenuating hepatocyte apoptosis and increasing insulin sensitivity. Exercise training also suppresses ROS overproduction and OS in NAFLD via up-regulation of several antioxidant enzymes and anti-inflammatory mediators. Therefore, an understanding of these molecules and signalling pathways gives us valuable information about NAFLD progression and a method for developing a suitable clinical treatment. This review aimed to evaluate sources of ROS and OS in NAFLD and the molecular mechanisms involved in the beneficial effects of exercises on NAFLD.     

长期运动训练对青年男性淋巴细胞凋亡及线粒体DNA修复酶的影响

目的探究长期运动训练对青年男性淋巴细胞凋亡及线粒体DNA(mtDNA)修复酶(8-氧鸟嘌呤DNA糖基化酶,OGG1)的影响及可能机制。方法训练组为20名男子足球专业运动员,对照组为20名男子大学生。2组均在功率自行车上完成递增负荷力竭运动。运动前和运动后即刻采集静脉血。流式细胞法检测淋巴细胞凋亡率,比色法检测超氧阴离子和羟自由基含量和Caspase-3,Caspase-9活性,高效液相色谱法检测mtDNA中8-氧鸟嘌呤(8-oxodG)含量,Western-blotting法检测线粒体OGG1蛋白(mtOGG1)表达水平。结果运动前和运动后,训练组与对照组比较,超氧阴离子、羟自由基、Caspase-3,Caspase-9活性、8-oxodG降低,OGG1升高;运动后,训练组与对照组比较,淋巴细胞凋亡率降低。对照组和训练组在运动后与运动前比较,淋巴细胞凋亡率、超氧阴离子、羟自由基、Caspase-3,Caspase-9活性和8-oxodG均升高,但训练组升高幅度低于对照组;运动后对照组OGG1降低,而训练组OGG1升高。结论一次性力竭运动诱导产生高水平ROS,并抑制mtOGG1表达,使mtDNA氧化损伤,促进淋巴细胞凋亡;长期耐力训练可通过抑制ROS生成,并提高mtOGG1表达,抑制运动性淋巴细胞凋亡。     

有氧运动中NADPH氧化酶介导产生的活性氧对大鼠神经胶质细胞活力的影响

建立大鼠有氧运动模型,原代培养了有氧运动后大鼠神经胶质细胞,阐明有氧运动后类NADPH氧化酶介导产生的活性氧(ROS)对细胞正常生理功能的影响。通过实验发现,有氧运动后大鼠神经胶质细胞中O2.-和H2O2生成均明显增加,而人工饲喂NADPH氧化酶的抑制剂二联苯碘(diphenylene iodonium,DPI)或夹竹桃麻素(apocynin,APO)后,均明显地降低由运动引起的大鼠神经胶质细胞O2.-和H2O2生成量;细胞活性的测定显示,DPI、APO、超氧化物歧化酶 过氧化氢酶均能明显降低运动后大鼠神经胶质细胞活性。表明有氧运动中类NADPH氧化酶介导产生的ROS是大鼠神经胶质细胞生存所必需,过度进补抗氧化剂将干扰细胞的正常生理活动甚至导致细胞死亡。     

过度运动激活中性粒细胞产生的活性氧对淋巴细胞DNA损伤及干预研究

目的:探讨过度运动激活中性粒细胞NADPH氧化酶介导产生的活性氧(ROS)对淋巴细胞DNA损伤的影响及实施二联苯碘(DPI)干预作用.方法:30只雄性Vistar大鼠随机分为3组,安静组(C,n=10)、过度训练组(E,n=10)和DPI干预组(D,n=10),E组和D组进行递增负荷跑台训练11周,末次训练结束后在36～40 h内各组随机取8只眼眶静脉采血.应用ELISA方法检测血浆细胞因子水平及血液中脂质过氧化水平;提取白细胞用AnnexinV/PI双染色法流式测定中性粒细胞凋亡与坏死程度;免疫细胞化学的激光共聚焦法测定NADPH氧化酶关键亚基的gp91 phox与p47phox的共定位;单细胞凝胶电泳测定淋巴细胞的DNA损伤程度.结果:1)与C组比较.E组血浆IL-1b显著性升高,IL-8、TNF-α显著升高(分别为P＜0.01和P＜0.05),MCP-1与CINC显著性下降(P＜0.05);而D组血浆IL-1b、MCP-1显著下降(P＜0.05),IL-8、TNF-α显著性升高(P＜0.05);E组和D组大鼠血浆慨、MPO水平均上升(分别为P＜0.01和P＜0.05);2)与C组相比,E组、D组中性粒细胞性凋亡的百分比均显著上升(P＜0.01),但中性粒细胞的死亡数E组增加(P＜0.05)而D组无显著变化;3)与C组比较,E组出现DNA损伤的彗星细胞数非常显著性升高(P＜0.01),且彗星细胞的宽度和尾长的变化均达到显著性水平,而D组的变化均未达到显著性水平;4)中性粒细胞的共聚焦染色定位显示:与安静组对照比较,E组出现了NADPH氧化酶关键亚基的gp91phox与p47phox的共定位.结论:1)过度运动可引起外周血炎性因子和趋化分子分泌的增加,从而有可能诱发组织炎症的发生和而调节T淋巴细胞的分化;2)过度运动可激活外周血中性粒细胞的NADPH氧化酶介导产生过多的ROS使血液的脂质过氧化水平提高;3)由此途径产生的ROS的增多和血液过氧化水平的提高可能引起中性粒细胞本身的凋亡甚至死亡和淋巴细胞的DNA损伤:4)由此途径产生的ROS的增多、吞噬细胞及淋巴细胞的过氧化损伤、炎性因子及免疫调节因子的变化等多因素可调节细胞免疫,是过度运动引起的运动型免疫抑制的可能因素.