高交感活性诱导的大鼠心肌损伤模型中氧化应激的受体调控机制*

 目的：研究高交感活性诱发大鼠心肌损伤的氧化应激受体调控机制。方法：Sprague-Dawley (SD)大鼠随机分为对照组、模型组、普萘洛尔（Pro) 组、哌唑嗪(Praz)组、普萘洛尔+哌唑嗪 (Pro+Praz) 组、维生素E(VE)组及普萘洛尔+哌唑嗪+维生素E (Pro+Praz+VE) 组,除对照组外其余各组均腹腔注射去甲肾上腺素(NE) 复制高交感活性引起的心肌损伤模型,同时灌胃给予相应药物,连续给药16 d,期间监测各组动物体重的变化。16 d后进行心室重构指标（心指数和羟脯氨酸含量）、病理组织学检查、氧化/抗氧化指标（MDA、SOD、CAT、GSH-Px和T-AOC）和能量代谢指标（Na+-K+ATPase和Ca2+-Mg2+ATPase）分析。结果：从第9天开始,模型组动物体重与对照组的比较差异有统计学意义（P<0.05）,心指数和左心室肥厚明显增加,氧化/抗氧化和能量代谢障碍;Pro、Praz、Pro+Praz和VE各组均出现不同程度的动物体重、心指数、左心室肥厚和氧化/抗氧化失衡的改善;Pro、Praz和Pro+Praz能明显升高左心室Na+-K+ATPase和Ca2+-Mg2+ATPase的活性,Pro+Praz作用最明显（P<0.05）。结论：肾上腺素受体依赖是高交感活性诱导心肌氧化应激损伤的重要途径。     

应激猪睾丸葡萄糖转运蛋白1和葡萄糖转运蛋白2的表达与定位

目的 探讨正常和热应激条件下,葡萄糖转运蛋白1（GLUT1）和葡萄糖转运蛋白2（GLUT2）在成年猪睾丸的表达和定位。 方法 性成熟长白公猪9头,随机分为3组。局部阴囊热刺激组（n=3）,用自制电热毯置阴囊42 ℃加热1 h;环境热应激组（n=3）,每天置于37～40 ℃猪舍环境3 h,连续7 d,每天于热处理结束后,将实验猪驱赶回21～25 ℃猪舍环境;对照组（n=3）,饲养在21～25 ℃猪舍环境。局部热刺激6 h后和环境热应激处理结束24 h后,手术摘除双侧睾丸。用Real-time PCR、Western blotting和免疫组织化学技术检测猪睾丸组织内GLUT1和GLUT2的表达。 结果 Real-time PCR和Western blotting结果显示,与对照组相比,环境热应激组GLUT1蛋白和mRNA的表达差异不显著,局部阴囊热刺激组GLUT1蛋白和mRNA表达显著升高;环境热应激组和局部阴囊热刺激组,GLUT2蛋白和mRNA表达均显著升高。免疫组织化学结果发现,热处理前后,GLUT1蛋白在曲精小管内定位于精母细胞和圆形精子细胞;环境热应激组GLUT1蛋白染色与对照组相比,无明显差异,局部阴囊热刺激后,GLUT1染色变深,表达升高。热处理前后,GLUT2蛋白在曲精小管定位于生精细胞和支持细胞,环境热应激和局部阴囊热刺激导致GLUT2染色变深,表达升高。 结论 葡萄糖转运蛋白GLUT1和GLUT2表达于猪睾丸曲精小管,环境高温和阴囊局部热刺激导致GLUT1和GLUT2在猪睾丸的表达水平改变,提示这两种葡萄糖转运蛋白在猪精子发生过程中发挥重要作用。     

Morphology of the mitochondria in heat shock protein 60 deficient fibroblasts from mitochondrial myopathy patients. Effects of stress conditions

We have described two mitochondrial (mt) myopathy patients with reduced activities of various mt enzymes associated with significantly decreased amounts of heat shock protein 60 (hsp60). Experimental evidence suggested that the lack of hsp60 was the primary defect. Since hsp60 is essential for the proper folding of enzyme subunits in the mt matrix a partial deficiency of this protein can explain the observed defects of the mitochondria. Here we report on morphological studies aimed at obtaining more insight into the relation between lack of hsp60 and pathological changes of the mitochondria. Under standard culture conditions mitochondria in the partially hsp60 deficient fibroblasts showed profound morphological aberrations. In contrast, the mitochondria in fibroblasts from a MELAS patient and a cytochrome c oxidase-deficient patient appeared normal. Under stress conditions the integrity of the hsp60 deficient mitochondria declined even further: heat shock induced a temporary collapse of the electrochemical potential across the inner mt membrane, but did not affect the ultrastructure of the mitochondria; prolonged growth in confluent cultures resulted in decrease in mt number. The altered mt morphology in the hsp60 deficient cells is probably indicative of the severely impaired mt metabolism whereas the decreased stress tolerance is likely to be a direct result of paucity of the heat shock protein. Both variables are potentially useful in the diagnosis and molecular characterization of mt disorders with systemic manifestation and multiple enzyme deficiency.     

Excess male mortality and age-specific mortality trajectories under different mortality conditions: a lesson from the heat wave of summer 2003

Introduction

Our objective was to study the impact of an identical additional stress on male and female mortality with a quasi-experimental study design, using natural variations in summer mortality, including the massive heat wave that struck Europe in 2003.

Material and Methods

The summer daily mortality rates of the population aged 65 and over living in 16 European countries were computed by single age from 1998 to 2003. Using the method of Tukey, we established five categories summarizing the summer daily conditions of mortality (exceptionally high values, minor extremely high values, common values, minor extremely low values, and exceptionally low values).

Results

Whatever the mortality conditions during the summer months, the mortality trajectories by age are exponential for both sexes: males die twice more than females at the age of 65 and their level of mortality linearly converges around the age of 97 to that of the females.

Discussion

Being male remains a major risk factor of mortality during heat waves. This issue was missed by previous epidemiological studies because almost all of them focused only on the relative increase in mortality and not on the sex specific mortality rates which implies being able to estimate the population at risk.     

Sleep-wake study in an animal model of acute and chronic heat stress

The study of the variations in different parameters of sleep-wake states following exposure to high environmental heat in three different age groups of freely moving rats have been presented in this paper. Each age group of rats was subdivided in three group (i) acute heat stress--subjected to a single exposure for 4 h in the BOD (Biological Oxygen Demand) incubator at 38 degrees C; (ii) chronic heat stress--exposed for 21 days daily for 1 h in the incubator at 38 degrees C, and (iii) handling control groups. The polygraphic, analog as well as digital sleep-wake recordings were performed just after the heat exposure from acute stressed rats and on 22nd day from chronic stressed rats. The results of this study revealed that acute exposure to high environmental heat increases sleep efficiency with significant increase in SWS (slow wave sleep) decrease in AWA (awake) time in all three age groups of rats. The increase in SWS and the sleep efficiency in these groups of rats at the cost of decreased time of AWA, indicates the involvement of the hypothalamus in thermoregulatory mechanism to conserve the energy of the body following sudden exposure to high heat. However, the reverse results were observed in the chronic stressed groups of rats, which have occurred mostly owing to the adaptations of the brain functions due to repetitive exposure to environmental heat. In consequence, the present study exhibits that the sleep is highly susceptible to the environmental heat and it is sensitive to the intensity, duration and the mode of exposure.     

阿托伐他汀通过抑制PKC的激活对抗高糖诱导的人脐静脉内皮细胞氧化应激反应

目的:探讨阿托伐他汀对高糖诱导的人脐静脉血管内皮细胞(HUVECs)产生氧化应激的影响及其作用机制。方法:体外培养HUVECs,以25 mmol/L葡萄糖干预,模拟糖尿病患者体内环境,通过流式细胞术和共聚焦显微镜检测细胞内的活性氧(ROS)水平,采用Lucigenin分析方法测定还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶活性,分别应用实时荧光定量PCR和免疫印迹杂交的方法检测 NADPH氧化酶亚基Nox4和Nox2/gp91^phox的表达水平,用免疫印迹杂交方法检测蛋白激酶C(PKC)蛋白的磷酸化水平。结果:(1)在高糖环境(终浓度为25 mmol/L)下,HUVECs内ROS生成显著增加,NADPH氧化酶的活性显著增强,NADPH 氧化酶Nox4和Nox2/gp91^phox亚基的mRNA和蛋白表达水平显著上调;(2)阿托伐他汀可显著抑制高糖诱导的ROS 生成、NADPH氧化酶活性的增强及NADPH 氧化酶Nox4和Nox2/gp91^phox亚基表达水平的增加幅度,且具有浓度依赖性;(3)PKC抑制剂(PKC inhibitor peptide, 20 μmol/L)可显著抑制高糖环境下ROS的生成、NADPH氧化酶活性的增强及NADPH 氧化酶Nox4和Nox2/gp91^phox亚基表达水平的增加幅度;(4)阿托伐他汀可抑制高糖诱导的PKC蛋白的磷酸化。结论:PKC的活化参与了高糖诱导的HUVECs产生的氧化应激反应。阿托伐他汀通过抑制PKC蛋白的活化对抗高糖诱导的内皮细胞产生的氧化应激反应。     

Combined effect of heat stress, dehydration and exercise on neuromuscular function in humans

This study examined the combined effect of exercise induced hyperthermia and dehydration on neuromuscular function in human subjects. Six trained male runners ran for 40 min on a treadmill at 65% of their maximal aerobic velocity while wearing a tracksuit covered with an impermeable jacket and pants to impair the evaporation of sweat. These stressful experimental running conditions led the runners to a physiological status close to exhaustion. On average, the 40 min run ended at a heart rate of 196 (SD 8) beats · min⁻¹, a tympanic temperature of 40 (SD 0.3) °C and with a loss of body mass of 2 (SD 0.5)%. Pre- and post-running strength tests included measurements of maximal knee extension and flexion torques in both isometric and isokinetic (at 60 and 240° · s⁻¹) conditions. A 20 s endurance test at 240° · s⁻¹ was also performed. Surface electromyographic (EMG) activity was recorded from six knee extensor and flexor muscles during the entire protocol. The treadmill run led to clear decrements in maximal extension torque and EMG activity both in isometric and at the slowest isokinetic velocity (60° · s⁻¹). However, no differences in these parameters were observed at 240° · s⁻¹. Furthermore, the EMG patterns of the major knee extensor and flexor muscles remained remarkably stable during the treadmill run. These results demonstrate that the exercise-induced hyperthermia and dehydration in the present experiments had only minor effects on the neuromuscular performance. However, it is also suggested that high internal body temperature per se could limit the production of high force levels. Accepted: 26 September 2000     

Thermoregulatory stress during rest and exercise in heat in patients with a spinal cord injury

Summary Twelve subjects with spinal cord injuries and four controls (all male) were exposed to heat while sitting at rest or working at each of three environmental temperatures, 30, 35 and 40°C, with a relative humidity of 50%. Exercise was accomplished at a load of 50 W on a friction-braked cycle ergometer which was armcranked or pedalled. Functional electrical stimulation of the legs was provided to the subjects with quadriplegia and paraplegia to allow them to pedal a cycle ergometer. The data showed that individuals with quadriplegia had the poorest tolerance for heat. As an example, in this group, accomplishing armcrank ergometry while working at an environmental temperature of 40°C resulted in an increase in aural temperature of 2°C in 30 min. The aural temperature of individuals with paraplegia working for the same length of time under the same conditions rose approximately 1°C. There was virtually no change in the aural temperature in the control subjects.     

The influence of heat stress on plasma volume and intravascular proteins in sedentary females

Summary 23 untrained female subjects underwent a standardised thermal dehydration. Plasma volume (PV), hematocrit (HCT), total intravascular protein concentration (TPC), albumin concentration (AC), total globulin concentration (TGC) were determined before, immediately, 90 min and 180 min after the heat stress. The intravascular protein masses (IPM) were calculated from PV and protein concentration. In comparison to men the loss of body weight and PV was smaller. Consequently the TPC does not increase to the same amount in men. A tendency for a diminution of IPM could be observed but this shift was not significant, compared to males. According to these findings the colloid osmotic capacity remained on a relatively high level. Opposite to men the IPM of globulins increased in females after thermal dehydration.This study was supported by the Bundesinstitut für Sportwissenschaft and the Deutsche Forschungsgemeinschaft     

Pinealectomy in ground squirrels: effect on behavioral and physiological responses to heat stress

Richardson's ground squirrels, 1 year after pinealectomy, showed altered behavioral and physiological responses to heat stress when compared to an intact group. Pinealectomized squirrels bar-pressed more often for a cool temperature reward in a hot environment. When deprived of behavioral control of the hot environment, pinealectomized squirrels increased their oxygen consumption, had a higher body temperature, and displayed signs of greater thermal stress, including death, as compared to intact animals. When the intact group was pinealectomized and the experiments were repeated, the difference in behavioral and physiological responses to heat stress were not as great as with the 1 year pinealectomized group. A speculative explanation of the pineal gland's influence on central and peripheral control of evaporative water loss and peripheral blood circulation are offered. The pineal gland may exert a subtle influence on heat transfer mechanisms and adaptations to thermal stress.     

Cardiovascular responses to heat stress and blood volume displacements during exercise in man

Summary Subjects exercised in the upright position at approximately 50% of maximal oxygen consumption in four situations: in 25 C air, in 45 C air [mean skin temperature ( _sk) 35 C], in 35 C water immersed to the level of the xiphoid process, and finally wearing a suit perfused with 35 C water. The water immersion prevented gravitational shifts of blood volume to the legs. In this situation the forearm blood flow (FBF) rose continually with increasing core temperature (T_es) in contrast to the attenuation in rise above 38 C T_es in 45 C air. The differences were significant above 38.6 C T_es in experiments in eight subjects. The effects of immersion on cardiac output (CO), stroke volume (SV), and heart rate (HR) were studied in five of the subjects in relation to T_es, since the rate of rise of T_es was different in the four situations. CO and SV tended to be higher during both rest and exercise in the water than in the other three conditions, while HR rose in the same manner with increasing core temperature, except that it was lower in 25 C air, where T_es was lower. Thus, the prevention of hydrostatic shifts of peripheral venous volume permitted the maintenance of a higher SV and peripheral blood flow, and enhanced the ability of the circulation to deal with the combined exercise and heat stress.Supported by the Danish Space Board (1112-32/81), the Danish Medical Research Council (512–15983), and the Danish Sports Research Council. Dr. Rowell was supported in part by NHLBI Grant HL 16910     

Relation between cardiac vagal tone and performance in male military personnel exposed to high stress: three prospective studies

Vagal tone has been proposed both as an index of emotion regulation and cognitive ability. To assess the relation between vagal tone and emotion regulation and cognitive ability, the present research prospectively measured vagal tone (measured either as high-frequency spectral power or respiratory sinus arrhythmia) in healthy participants exposed to high stress. The participants were active duty military personnel (men) enrolled in high intensity military training: Survival School (Experiments 1 and 3) and Combat Diver Qualification Course (Experiment 2). We consistently observed a significant relationship between low vagal tone and superior performance. The data suggest that vagal suppression is associated with enhanced performance under conditions of high stress and that this enhanced performance may be related to emotion regulation and cognitive functioning.     

Differential regulation and activity against oxidative stress of Dps proteins in Bacillus cereus

In this study, the sequence similarity, structure, ferroxidase activity and efficacy in antagonizing oxidative stress of three Dps-like proteins, Dps1, Dps2 and Dps3, encoded by Bacillus cereus were comparatively analyzed. The three Dps-like proteins are homologous to other bacterial Dps proteins that exhibit ferroxidase activity. Both Dps1 and Dps2 have a typical Dps spherical structure, but Dps3 has a unique filamentous structure. Several dps mutant strains were generated to investigate the functional role of dps genes in cell protection. The dps1 null strain was the most labile to oxidative stress in the stationary phase, and the loss of dps2 resulted in greater sensitivity to peroxide exposure compared with the other mutant strains in the log phase. Interestingly, after simultaneous deletion of dps1 and dps2, the survival rate was dramatically reduced by approximately 5 log in the stationary phase. Immunoblotting analysis demonstrated that Dps1 and Dps2 in the wild-type strain were induced by oxidative stress, and Dps3 responded to general stress in the log phase. Constitutively high expression of Dps2 in a perR null mutant and PerR-specific binding of the promoter region of dps2 confirmed Dps2 as a member of the PerR regulon. In addition, the expression of Dps1 and Dps2, absent any stress, was initiated in the log phase and was abundant in the stationary phase, suggesting that the expression of Dps1 and Dps2 was dependent on the bacterial growth stage. In summary, the three Dps proteins conferred cellular protection, particularly from oxidative stress, and were differentially regulated in response to varied stress conditions.     

Serum protein peroxidation in rats selected by the rate of active avoidance conditioning under normal conditions and during stress

Serum protein peroxidation was studied in rats with different strategy of adaptive behavior at rest and during short-term immobilization stress. Oxidative destruction of serum proteins was less intensive in KLA rats compared to that in KHA rats. Stress stimulated peroxidation of serum proteins in KHA rats.