Extracellular Hsp72 concentration relates to a minimum endogenous criteria during acute exercise-heat exposure

Extracellular heat shock protein 72 (eHsp72) concentration increases during exercise-heat stress when conditions elicit physiological strain. Differences in severity of environmental and exercise stimuli have elicited varied response to stress. The present study aimed to quantify the extent of increased eHsp72 with increased exogenous heat stress, and determine related endogenous markers of strain in an exercise-heat model. Ten males cycled for 90 min at 50 % in three conditions (TEMP, 20 °C/63 % RH; HOT, 30.2 °C/51%RH; VHOT, 40.0 °C/37%RH). Plasma was analysed for eHsp72 pre, immediately post and 24-h post each trial utilising a commercially available ELISA. Increased eHsp72 concentration was observed post VHOT trial (+172.4 %) (p < 0.05), but not TEMP (−1.9 %) or HOT (+25.7 %) conditions. eHsp72 returned to baseline values within 24 h in all conditions. Changes were observed in rectal temperature (T_rec), rate of T_rec increase, area under the curve for T_rec of 38.5 and 39.0 °C, duration T_rec ≥ 38.5 and ≥39.0 °C, and change in muscle temperature, between VHOT, and TEMP and HOT, but not between TEMP and HOT. Each condition also elicited significantly increasing physiological strain, described by sweat rate, heart rate, physiological strain index, rating of perceived exertion and thermal sensation. Stepwise multiple regression reported rate of T_rec increase and change in T_rec to be predictors of increased eHsp72 concentration. Data suggests eHsp72 concentration increases once systemic temperature and sympathetic activity exceeds a minimum endogenous criteria elicited during VHOT conditions and is likely to be modulated by large, rapid changes in core temperature.     

Moderate- and high-intensity exhaustive exercise in the heat induce a similar increase in monocyte Hsp72

This study examined the relationship between exhaustive exercise in the heat at moderate and high intensities on the intracellular heat shock protein 72 (iHsp72) response. Twelve male subjects cycled to exhaustion at 60 and 75 % of maximal oxygen uptake in hot conditions (40 °C, 50 % RH). iHsp72 concentration was measured in monocytes before, at exhaustion and 24 h after exercise. Rectal temperature, heart rate and oxygen uptake were recorded during exercise. Volitional exhaustion occurred at 58.9 ± 12.1 and 27.3 ± 9.5 min (P < 0.001) and a rectal temperature of 39.8 ± 0.4 and 39.2 ± 0.6 °C (P = 0.002), respectively, for 60 and 75 %. The area under the curve above a rectal temperature of 38.5 °C was greater at 60 % (17.5 ± 6.6 °C min) than 75 % (3.4 ± 4.8 °C min; P < 0.001), whereas the rate of increase in rectal temperature was greater at 75 % (5.1 ± 1.7 vs. 2.2 ± 1.4 °C h⁻¹; P < 0.001). iHsp72 concentration increased similarly at exhaustion relative to pre-exercise (P = 0.044) and then increased further at 24 h (P < 0.001). Multiple regression analysis revealed no predictor variables associated with iHsp72 expression; however, a correlation was observed between exercise intensities for the increase in iHsp expression at exhaustion and 24 h (P < 0.05). These results suggest that iHsp72 expression increased in relation to the level of hyperthermia attained and sustained at 60 % and the higher metabolic rate and greater rate of increase in core temperature at 75 %, with the further increase in iHsp72 concentration 24 h after exercise reinforcing its role as a chaperone and cytoprotective agent.     

Induction of Mesenchymal Stem Cells Leads to HSP72 Synthesis and Higher Resistance to Oxidative Stress

The phenomenon of neuronal transdifferentiation performed on bone marrow mesenchymal stem cells (MSCs) has been criticized by recent studies indicating that acquired neuron-like morphology of induced MSCs is caused by cellular stress. Therefore, to test this hypothesis we have investigated whether exposure of rat MSCs (rMSCs) to chemical inducer 2 mM β-mercaptoethanol (BME) for 1–3 h followed by 24 h incubation leads to HSP72 synthesis, thus suggesting higher resistance of rMSCs to oxidative damage. Present data from immunohistochemistry clearly indicate development of time-dependent sub-cellular HSP72 distribution, initially seen in nuclei at 1 h followed by its translocation to surrounding central cytoplasm and processes at 2–3 h after BME stimulation. Western blot (WB) analysis confirmed the expression of HSP72 protein in induced rMSCs at both stimulation periods. Furthermore, preconditioned rMSCs with BME for 1 h expressing HSP72 positivity at 24 h showed higher resistance (78 ± 10% of survival cells) to oxidative stress caused by 1 mM H₂O₂ when compared to those preconditioned for 3 h (59 ± 8% of survival cells) or control-unconditioned rMSCs exposed to the same stressor conditions (56 ± 6% of survival cells). Thus, the cellular protection was lost if the duration of BME preconditioning was increased as far as possible (3 h) (while still remaining sub-lethal). This suggests that exposure of rMSCs to the optimal concentration of BME (2 mM) during optimal induction period (1 h) mediate their protection and increases resistance to oxidative injury, while over crossing these limits is in-effective. In addition, our findings confirm that cultured rMSCs remain competent to be preconditioned by BME, through a pathway that may increase the antioxidant balance or involve activation of HSP72 protein induced tolerance.     

Lymphocyte HSP72 following exercise in hyperthermic runners: The effect of temperature

Heat shock protein 72 (HSP72) is the most inducible HSP, but is not always increased in lymphocytes following exercise. This field study examined whether lymphocyte HSP72 was increased in hyperthermic (T_rec>39.0 °C) male athletes following a 14 km competitive race in cool conditions (ambient temperature 11.2 °C). A comparison was also made between control runners (n=7) and those treated for exertional heat illness (n=9). Lymphocyte HSP72 was not increased in control runners immediately post- compared with pre-race, and there was no difference between both groups of runners. A second study of the race (ambient temperature 14.6 °C) found that lymphocyte HSP72 in control (n=7) and treated (n=9) athletes was higher 2 days post- compared with immediately post-race (p<0.01) and these increases were correlated with post-exercise T_rec (p<0.05).     

热射病合并多器官功能障碍综合征持续静-静脉血液滤过治疗的护理

目的：探讨热射病合并多器官功能障碍综合征（MODs）持续静-静脉血液滤过（CVVH）的治疗作用并总结护理要点。方法：回顾分析2008年5月至2010年9月,我院收治的6例热射病合并MODS,及时运用CVVH并采取综合治疗措施,对治疗和护理进行总结。结果：6例患者5例痊愈,1例死亡。通过早期积极采用CVVH联合综合治疗措施,可快速降低患者体温,防止因高热引起的恶性循环。结论：CVVH是热射病合MODS有效治疗方法,其可早期纠正电解质及酸碱紊乱,且对代谢产物和炎症因子的清除也有重要作用。期间严密的观察与护理,以及配合治疗的连续性,是成功救治这类患者的关键。     

Induction of stress proteins by sodium chloride treatment in Bacillus subtilis

In Bacillus subtilis, heat shock proteins can be classified into two main groups: specific heat shock proteins (about 5) and general stress proteins (at least 14). Salt stress was very effective in the induction of general stress proteins (5 to 50-fold stimulation), but the synthesis of heat-specific stress proteins was not stimulated. Furthermore there were some proteins whose synthesis was accelerated only by salt stress.     

热射病合并多器官功能障碍综合征持续静- 静脉血液滤过治疗的护理

目的:探讨热射病合并多器官功能障碍综合征(MODS)持续静-静脉血液滤过(CVVH)的治疗作用并总结护理要点。方法:回顾分析2008年5月至2010年9月,我院收治的6例热射病合并MODS,及时运用CVVH并采取综合治疗措施,对治疗和护理进行总结。结果:6例患者5例痊愈,1例死亡。通过早期积极采用CVVH联合综合治疗措施,可快速降低患者体温,防止因高热引起的恶性循环。结论:CVVH是热射病合MODS有效治疗方法,其可早期纠正电解质及酸碱紊乱,且对代谢产物和炎症因子的清除也有重要作用。期间严密的观察与护理,以及配合治疗的连续性,是成功救治这类患者的关键。     

Heat and exercise acclimation increases intracellular levels of Hsp72 and inhibits exercise-induced increase in intracellular and plasma Hsp72 in humans

In order to verify the effects of heat and exercise acclimation (HA) on resting and exercise-induced expression of plasma and leukocyte heat shock protein 72 (Hsp72) in humans, nine healthy young male volunteers (25.0 ± 0.7 years; 80.5 ± 2.0 kg; 180 ± 2 cm, mean ± SE) exercised for 60 min in a hot, dry environment (40 ± 0°C and 45 ± 0% relative humidity) for 11 days. The protocol consisted of running on a treadmill using a controlled hyperthermia technique in which the work rate was adjusted to elevate the rectal temperature by 1°C in 30 min and maintain it elevated for another 30 min. Before and after the HA, the volunteers performed a heat stress test (HST) at 50% of their individual maximal power output for 90 min in the same environment. Blood was drawn before (REST), immediately after (POST) and 1 h after (1 h POST) HST, and plasma and leukocytes were separated and stored. Subjects showed expected adaptations to HA: reduced exercise rectal and mean skin temperatures and heart rate, and augmented sweat rate and exercise tolerance. In HST1, plasma Hsp72 increased from REST to POST and then returned to resting values 1 h POST (REST: 1.11 ± 0.07, POST: 1.48 ± 0.10, 1 h POST: 1.22 ± 0.11 ng mL⁻¹; p < 0.05). In HST2, there was no change in plasma Hsp72 (REST: 0.94 ± 0.08, POST: 1.20 ± 0.15, 1 h POST: 1.17 ± 0.16 ng mL⁻¹; p > 0.05). HA increased resting levels of intracellular Hsp72 (HST1: 1 ± 0.02 and HST2: 4.2 ± 1.2 density units, p < 0.05). Exercise-induced increased intracellular Hsp72 expression was observed on HST1 (HST1: REST, 1 ± 0.02 vs. POST, 2.9 ± 0.9 density units, mean ± SE, p < 0.05) but was inhibited on HST2 (HST2: REST, 4.2 ± 1.2 vs. POST, 4.4 ± 1.1 density units, p > 0.05). Regression analysis showed that the lower the pre-exercise expression of intracellular Hsp72, the higher the exercise-induced increase (R = −0.85, p < 0.05). In conclusion, HA increased resting leukocyte Hsp72 levels and inhibited exercise-induced expression. This intracellular adaptation probably induces thermotolerance. In addition, the non-increase in plasma Hsp72 after HA may be related to lower stress at the cellular level in the acclimated individuals.     

肠道菌群及内毒素在多器官功能不全综合征时的变化

目的　探讨肠道菌群及内毒素在多器官功能不全综合征( MODS)时的变化。方法　取SD大鼠,腹腔注射无菌酵母多糖A制备MODS模型,检测大鼠肠道菌群、外周血和门静脉血中的内毒素以及肠道游离内毒素含量,并进行定量分析。结果　模型组大鼠肠道专性厌氧菌的数量明显减少,革兰阴性杆菌和双歧杆菌的比例倒置,内毒素含量明显增加,与对照组比差异有显著性( P<0 .0 5 )。结论　MODS时肠道细菌微生态发生明显改变,肠道内毒素池与肠道革兰阴性杆菌的变化密切相关     

Role of HSF activation for resistance to heat, cold and high-temperature knock-down

Regulation of heat shock proteins (Hsps) by the heat shock factor (HSF) and the importance of these proteins for resistance to heat stress is well documented. Less characterized is the importance of Hsps for cold stress resistance although Hsp70 is known to be induced following long-term cold exposure in Drosophila melanogaster. In this study, a temperature-sensitive HSF mutant line was used to investigate the role of HSF activation following heat hardening, rapid cold hardening (RCH) and long-term cold acclimation (LTCA) on heat and cold resistance, and this was correlated with Hsp70 expression. In addition, the effect of HSF activation on high-temperature knock-down resistance was evaluated. We found a significantly decreased HSF activation in the mutant line as compared to a corresponding control line following heat hardening, and this was correlated with decreased heat resistance of the mutant line. However, we did not find this difference in HSF activity to be important for resistance to cold stress or high-temperature knock-down. The findings indicate that induction of stress genes regulated by HSF, such as Hsps, although occurring following LTCA, are not of major importance for cold stress resistance and neither for RCH nor high-temperature knock-down resistance in D. melanogaster.     

Alterations of trace elements (Zn, Se, Cu, Fe) and related metalloenzymes in rabbit blood after severe trauma

To investigate the status of the trace elements (TEs) and related metalloenzymes activities in the injury and repair process after severe trauma, we established a rabbit model of severe trauma whose Injury Severity Score (ISS) was 22. Concentrations of blood selenium (Se) and serum copper (Cu), zinc (Zn), iron (Fe), and ferritin were measured on D0 (before injury), and day (D) 1, D2, D3, D6, D9, D14, D21, D28 after trauma, respectively. The activities of glutathione peroxidase (GPx), Cu/Zn superoxide dismutase (Cu/Zn-SOD), myeloperoxidase (MPO), the contents of lipid peroxidation product malondialdehyde (MDA) and serum biochemical profile were detected synchronously. In addition, the morphologic changes of major organs were observed at different time intervals. Results showed that blood Se and serum Zn, Fe contents decreased significantly within 2 weeks after injury. Serum Cu concentration was significantly reduced on D1 but normalized quickly. Serum ferritin level increased during the first week while following an obvious decrease thereafter. The blood GPx activity dropped markedly from D1 to D6, the serum Cu/Zn-SOD activity decreased on D1 and then increased significantly within 2 weeks, and the blood MPO-positive stained cells increased within a week after trauma and followed by a decrease from D14 to D21. The serum MDA increased significantly on D6. Seven of 34 rabbits died in 4-6 days after injury. Biochemistry values and pathological features revealed these rabbits died of multiple organ dysfunction syndrome (MODS). Our experiment suggested that the circulating TEs status is dramatically modified in response to trauma, which might be a factor in MODS.     

Effects of prolonged stanozolol treatment on antioxidant enzyme activities, oxidative stress markers, and heat shock protein HSP72 levels in rat liver

The abuse of anabolic-androgenic steroids (AAS) to enhance physical performance is widespread in sport communities despite their reported side effects. Since the biochemical bases for the hepatotoxic effects of these compounds are largely unknown, this investigation was aimed at testing whether prolonged (8 weeks) treatment with high doses (2 mg kg⁻¹ body weight; 5 d wk⁻¹) of stanozolol (ST), either alone or in conjunction with treadmill-exercise training, induced changes in oxidative stress biomarker levels and antioxidant defence systems in rat liver. After ST oral administration, the mean values of serum parameters related to hepatic function were within normal ranges. No changes in protein carbonyl content and in the reduced to oxidized glutathione (GSH/GSSG) ratio were detected in liver homogenates of ST-treated rats, whereas thiobarbituric acid-reactive substances (TBARS) levels resulted increased (P<0.05). Total superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities were higher (P<0.05) in the liver of treated rats but mitochondrial SOD and glutathione reductase (GR) activities, and the 72 kDa heat shock protein (HSP72) level were not modified. Chronic exercise alone did not change any of the above parameters except for a remarkable enhancement of HSP72 expression; in no case training modified the effects of ST treatment. The present data show that 8 wk ingestion of ST, either with or without concurrent exercise training, can induce oxidative stress in rat liver despite the up-regulation of enzymatic antioxidant activities.     

Expression of heat shock proteins in adult honey bee (Apis mellifera L.) workers under hot-arid subtropical ecosystems

Heat stress elicits the expression of heat shock proteins (HSPs) in honey bee subspecies. These highly conserved proteins have significant role in protecting cells from thermal-induced stresses. Honey bees in subtropical regions face extremely dry and hot environment. The expression of HSPs in the nurses and foragers of indigenous (Apis mellifera jemenitica) and imported European (Apis mellifera ligustica and Apis mellifera carnica) honey bee subspecies after heat shock treatment were compared using SDS-PAGE. Hsp70 and Hsp82 were equally expressed in the nurses of all tested bee subspecies when exposed to 40 °C and 45 °C for 4 h. The forager bees exhibited differential expression of HSPs after heat stress. No HSPs was expressed in the foragers of A. m. jemenitica, and Hsp70 was expressed only in the foragers of A. m. ligustica and A. m. carnica at 40 °C. A prominent diversity in HSPs expression was also exhibited in the foragers at 45 °C with one HSP (Hsp70) in A. m. jemenitica, two HSPs (Hsp40 and Hsp70) in A. m. carnica, and three HSPs (Hsp40, Hsp60 and Hsp70) in A. m. ligustica. No HSPs was expressed in the control nurse and forager bees at any of the tested temperatures. These findings illustrated the differences in HSP expression among nurse and forager bees. It is obvious that the native foragers are more heat tolerant with least HSPs expression than exotic bee races. Further investigations will help to understand the potential role of HSPs in the adaptability, survival, and performance of bee subspecies in harsh climate of the subtropical regions.     

不同血液净化方式对中毒所致多器官功能障碍的临床效果比较

目的:探讨两种不同的血液净化方式对中毒所致的多器官功能障碍综合征(multiple organ dysfunction syndrome,MODS)的临床治疗效果。方法:回顾性分析2015年9月至2018年9月我院收治的41例中毒所致的多器官功能障碍综合征患者,按照采取血液净化方式的不同将其分为实验组(序贯性净化)和对照组(单种净化)。比较两组患者在住院期间的一般住院指标(住院时间、费用等)及治疗后的主要血液指标、APACHE Ⅱ (acute physiology and chronic health evaluationⅡ)评分和生存率。结果:治疗后第5天,实验组的丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)等生化指标显著低于对照组(P0.05)。治疗第5天时,实验组的APACHE Ⅱ评分显著低于对照组(P0.05),总住院费用高于对照组(P0.05),总住院时间及病死率与对照组比较差异无统计学差异(P0.05)。结论:相对于单种血液净化方式而言,序贯性治疗可显著改善肝功指标,降低APACHE Ⅱ评分,且不增加总住院时间,但增加了治疗费用。     

乌司他丁联合血必净对内毒素致绵羊多脏器功能障碍综合征的防治研究

目的探讨乌司他丁（UTI）联合血必净（XBJ）对内毒素（LPS）致多脏器功能障碍综合征（MODS）的防治作用。方法将50只绵羊随机分为对照组（A组）、模型组（B组）、乌司他丁组（C组）、血必净组（D组）和乌司他丁＋血必净组（E组）,每组10只。利用LPS建立MODS模型,并以UTI、XBJ和UTI＋XBJ进行防治。观察各组的体征表现,测定不同时点各组的血气、血常规、血清生化指标的改变。结果 UTI＋XBJ组绵羊的体征表现明显减轻、死亡率明显降低,PaO2、生化指标和白细胞数目得到了有效的控制。结论乌司他丁联合血必净较单独应用能明显改善LPS诱导的MODS动物体征表现、降低死亡率,对肺脏、肝脏和肾脏等多脏器损伤具有良好的防治作用。     

Development of the embryonic heat shock response and the impact of repeated thermal stress in early stage lake whitefish (Coregonus clupeaformis) embryos

Lake whitefish (Coregonus clupeaformis) embryos were exposed to thermal stress (TS) at different developmental stages to determine when the heat shock response (HSR) can be initiated and if it is altered by exposure to repeated TS. First, embryos were subject to one of three different TS temperatures (6, 9, or 12 °C above control) at 4 points in development (21, 38, 60 and 70 days post-fertilisation (dpf)) for 2 h followed by a 2 h recovery to understand the ontogeny of the HSR. A second experiment explored the effects of repeated TS on the HSR in embryos from 15 to 75 dpf. Embryos were subjected to one of two TS regimes; +6 °C TS for 1 h every 6 days or +9 °C TS for 1 h every 6 days. Following a 2 h recovery, a subset of embryos was sampled. Our results show that embryos could initiate a HSR via upregulation of heat shock protein 70 (hsp70) mRNA at all developmental ages studied, but that this response varied with age and was only observed with a TS of +9 or +12 °C. In comparison, when embryos received multiple TS treatments, hsp70 was not induced in response to the 1 h TS and 2 h recovery, and a downregulation was observed at 39 dpf. Downregulation of hsp47 and hsp90α mRNA was also observed in early age embryos. Collectively, these data suggest that embryos are capable of initiating a HSR at early age and throughout embryogenesis, but that repeated TS can alter the HSR, and may result in either reduced responsiveness or a downregulation of inducible hsps. Our findings warrant further investigation into both the short- and long-term effects of repeated TS on lake whitefish development.     

连续静脉-静脉血液透析滤过治疗多器官功能障碍综合征的临床疗效研究

目的:探讨连续静脉-静脉血液透析滤过(continuous veno-venous hemodialysis filtration,CVVHDF)治疗多器官功能障碍综合征(MODS)的临床疗效。方法:选择我院ICU自2015年1月-2017年2月收治的MODS患者50例作为研究对象,根据随机抽签原则分为观察组与对照组,每组25例。对照组在常规内科治疗基础上给予机械通气治疗,观察组在对照组基础上采用CVVHDF治疗,两组治疗观察时间为30d。观察和比较两组ICU停留时间、呼吸机辅助时间、治疗后APACHEⅡ评分与MODS评分、治疗前后血清IL-6与TNF-α含量的变化及随访6个月的死亡情况。结果:治疗后,观察组ICU停留时间和呼吸机辅助时间均短于对照组(P0.05),APACHEⅡ评分与MODS评分分别为22.33±2.49分和6.42±1.98分,均明显低于对照组(62.19±7.45分和7.29±1.67分)(P0.05)。治疗后,两组的血清IL-6与TNF-α含量都明显低于治疗前,且观察组的血清IL-6与TNF-α含量均明显低于对照组(P0.05)。随访6个月,观察组与对照组的死亡率分别为4.0%和20.0%,观察组的死亡率低于对照组(P0.05)。结论:CVVHDF治疗MODS能有效清除患者炎症因子,缩短ICU停留时间和呼吸机辅助时间,降低患者的死亡率。     

A microarray analysis of the effects of moderate hypothermia and rewarming on gene expression by human hepatocytes (HepG2)

The gene expression changes produced by moderate hypothermia are not fully known, but appear to differ in important ways from those produced by heat shock. We examined the gene expression changes produced by moderate hypothermia and tested the hypothesis that rewarming after hypothermia approximates a heat-shock response. Six sets of human HepG2 hepatocytes were subjected to moderate hypothermia (31°C for 16 h), a conventional in vitro heat shock (43°C for 30 min) or control conditions (37°C), then harvested immediately or allowed to recover for 3 h at 37°C. Expression analysis was performed with Affymetrix U133A gene chips, using analysis of variance-based techniques. Moderate hypothermia led to distinct time-dependent expression changes, as did heat shock. Hypothermia initially caused statistically significant, greater than or equal to twofold changes in expression (relative to controls) of 409 sequences (143 increased and 266 decreased), whereas heat shock affected 71 (35 increased and 36 decreased). After 3 h of recovery, 192 sequences (83 increased, 109 decreased) were affected by hypothermia and 231 (146 increased, 85 decreased) by heat shock. Expression of many heat shock proteins was decreased by hypothermia but significantly increased after rewarming. A comparison of sequences affected by thermal stress without regard to the magnitude of change revealed that the overlap between heat and cold stress was greater after 3 h of recovery than immediately following thermal stress. Thus, while some overlap occurs (particularly after rewarming), moderate hypothermia produces extensive, time-dependent gene expression changes in HepG2 cells that differ in important ways from those induced by heat shock.     

Hyperthermia Induces the Synthesis of a Heat Shock Protein by Polysomes Isolated from the Fetal and Neonatal Mammalian Brain

Abstract: Analysis of the cell-free translation products of polysomes isolated from fetal brain and other organs indicates that elevation of maternal body temperature induces the synthesis of a heat shock protein of molecular weight 74,000 (74K). The newborn mammal is particularly sensitive to induction of the 74K protein. As early postnatal development proceeds, higher body temperatures are required to induce synthesis of the 74K heat shock protein.