鸡热应激与糖皮质激素受体和热休克蛋白的相关性

本研究应用３ Ｈ Ｄｅｘ 放射配体结合的 Ｓｃａｔｃｈａｒｄ 分析和３５ Ｓ蛋氨酸体外标记法分别测定了环境温度 ４０ ℃热应激时,鸡外周血淋巴细胞（ Ｐ Ｂ Ｌ）的糖皮质激素受体（ Ｇ Ｒ）的最大结合容量（ Ｒｏ）、平衡解离常数（ Ｋｄ）及其热休克蛋白的表达。结果显示在热应激处理后０．５～４ ｈ 鸡 Ｐ Ｂ Ｌ、 Ｇ Ｒ、 Ｒｏ,从（８５．７５±１０．０９） ｆｍ ｏｌ／１０７ 细胞迅速下降至（１６．３４±２．８９） ｆｍ ｏｌ／１０７ 细胞,仅为正常对照组 Ｒｏ 的１９．０６％ （ Ｐ＜ ０．０１）。与此同时细胞热休克蛋白（ Ｈ Ｓ Ｐ）的合 成持续增加,主要有 Ｈ Ｓ Ｐ９０, Ｈ Ｓ Ｐ７０ 和 Ｈ Ｓ Ｐ２５。而在热应激最初０．５ ｈ 细胞总的蛋白合成急剧下降,随着应激时间的延长, Ｈ Ｓ Ｐ合成的增多,细胞总的蛋白也逐渐恢复。揭示 Ｈ Ｓ Ｐ在热应激过程中对细胞结构和机能的重建、维持激素与 Ｇ Ｒ 的亲合力,稳定受体蛋白结构,提高机体热耐受力具有重要意义。     

热休克蛋白对胚胎早期发育的影响

热休克现象是1962年R itossa首先在果蝇幼虫动物试验中发现的。试验发现,果蝇唾液腺染色体在过热环境中会发生形态改变。此现象与细胞中一类特殊蛋白质———热休克蛋白(Heat shock protein,HSP)的合成有关,因其最先在热休克现象中发现而得以命名。热休克蛋白除了因热休克刺激产生以外,还有很多因素(如缺氧、低温、氧化反应、化学物质、各种重金属、放射线、某些线粒体呼吸链抑制剂及某些药物等)都可以诱导产生,因此热休克蛋白又称为应激蛋白质(stress protein)。HSP70和HSP90是分子质量分别在70,90 ku左右的热休克蛋白,是热休克蛋白家…     

热休克蛋白70研究新进展

1962年,Ritassa在果蝇的研究中首次发现,短暂的热休克可以诱导唾液腺染色体出现3个膨突,提示这一区带转录加强,他将这一现象称为“热休克反应“(heat shock response,HSR).1974年,Tissieres发现热休克反应中转录合成的为一组特殊蛋白,而且伴随着这类蛋白的合成,细胞的其他蛋白合成却受到抑制,由于这类蛋白的合成与热休克反应有关,故命名为热休克蛋白(heat shock protein,HSP).除了高热之外,多种应激原如重金属、饥饿、缺氧、缺血等都可以诱导HSP的表达,但人们习惯上仍称为HSP或热应激蛋白(heat stress protein,HSP),有时也称为应激蛋白(stress protein,SP).……     

热应激雏鸡免疫器官中热休克蛋白70的定位研究

通过检测热休克蛋白70（heat shock protein,HSP70）在免疫器官及细胞上的分布规律,研究热应激雏鸡免疫器官中热休克蛋白70的定位及表达。结果显示：HSP70在常温饲养雏鸡的胸腺、脾脏、法氏囊中均有明显的表达,在胸腺、脾脏的细胞中主要以胞浆表达为主,而法氏囊中则以胞核表达为主,脾脏中HSP70的表达强度最为稳定。热应激时,胸腺中HSP70的IOD值随日龄增长而逐渐降低,末期显著低于对照组（P〈0．05）;脾脏中HSP70的IOD值随日龄增长逐渐降低,末期稍低于对照组（P〉0．05）;法氏囊中HSP70的IOD值与饲养日龄间无明显规律,末期高于对照组（P〉0．05）。结果表明,热应激对雏鸡胸腺的损伤明显,胸腺细胞通过自身合成HSP70抵抗热应激的能力较差,而对脾脏和法氏囊影响较小。     

热应激蛋白70与细胞凋亡

热应激蛋白 (heatstressprotein,HSP)或热休克蛋白 (heatshockpro tein,HSP)是机体受到应激原的刺激后产生的几族蛋白质 ,具有高度保守性 ,对维持细胞生存和内环境稳定起重要作用 [1]。多种应激原如高热、重金属、饥饿、缺氧、缺血等都可诱导HSP的表达 ,但人们习惯上仍称其为热应激蛋白或热休克蛋白 ,有时也称为应激蛋白 (stressprotein,SP)。一般来说 ,根据HSP的同源性、功能和分子量 ,主要HSP可分成4个家族 :小分子量HSP家族、HSP70家族、HSP90家族和大分子量HSP家族。HSP70家族是HSP中最保守和最重要的一族 ,在大多数生物…     

热休克蛋白90在糖皮质激素受体信号通路中的分子伴侣作用

在应激反应时,下丘脑-垂体-肾上腺轴(HPA)在机体的调节方面起主要作用,而糖皮质激素(GC)是HPA轴中的关键物质,因此在考量急性和慢性应激反应时,GC及其受体(GR)的变化常常是首选的研究内容.而GR正常构象的形成、激活、功能发挥与其分子伴侣-热休克蛋白90 (HSP90)关系密切.综述了HSP90的结构和功能、GC-GR信号通路中GC和GR的作用机制以及HSP90在GR信号通路中的分子伴侣作用.     

热休克蛋白的研究进展

热休克蛋白(Heatshock proteins,HSP),是生物受到环境中物理、化学、生物、精神等刺激时产生应激反馈而合成的蛋白质。作为进化保守的蛋白家族之一,普遍存在于各种生物体中,并在生物体内发挥着重要的生理功能。HSP因具有丰富强大的分子伴侣、抗应激、调节细胞凋亡、抗氧化、参与机体免疫等生物学功能,决定了其广泛的用途。大量的试验表明,近年来HSP在相关领域的应用研究已获得了重要的突破与进展,特别是在疾病免疫和药物开发中的应用研究。文中在阐明HSP分子调控的基础上,综述了近年来热休克蛋白家族(HSP90、HSP70、HSP60和小分子HSP)在功能和应用方面的研究进展,为进一步完善对HSP的研究提供一定的参考。     

HSP在热应激条件下调控鸡心肌细胞损伤的功能分析

心肌对热敏感,心脏成为热应激损伤的重要器官之一,严重时会引发猝死。热休克蛋白(HPS)是机体在遭受热应激刺激时激活的重要内源性保护机制。为了观察热应激条件下对蛋鸡心肌组织的病理性损伤,以及心肌细胞启动热休克蛋白的变化规律,选用40只300日龄海兰褐蛋鸡构建热应激试验动物模型,通过对血清中肌酸激酶(CK)、肌酸激酶同工酶MB(CK-MB)、乳酸脱氢酶(LDH)和谷草转氨酶(AST)水平的检测以及病理学检测以确定热应激损伤;通过对心肌组织的Hsps的转录水平检测以确定对Hsps的激活;通过系统网络分析,筛选出调控家禽热应激细胞凋亡的相关靶标,并进行相互作用网络和蛋白共表达分析。研究表明,38℃热应激会引起蛋鸡心肌细胞颗粒变性、空泡变性和坏死,导致心肌损伤相关酶谱CK、CK-MB、LDH和AST的上调,还可以激活Cryab、Hsp60、Hsp70和Hsp90转录水平的升高。系统网络分析发现热休克蛋白家族成员CRYAA、HSP90AB1、HSP90B1、HSPA5、HSPD1、DNAJA1和DNAJC3均参与热应激条件下调控家禽细胞凋亡的进程,Hsp90AB1和Hsp90B1可以调控CDK1,Hsp90AB1和HspD1可以与CDK1共表达,Hsp90B1、DNAJC3和IGF1可以调控IL6,影响细胞周期,从而影响细胞凋亡。     

热休克蛋白70在热应激鸡原代心肌细胞中的表达与定位

为研究热应激后鸡原代心肌细胞中热休克蛋白70(HSP70)的表达量及其定位的变化,试验将体外培养的鸡原代心肌细胞分为6组,其中1组作为对照(未进行热应激处理),其余5组于42℃热应激处理0.5,1,2,4,8 h,Western-blot检测HSP70在6组鸡原代心肌细胞中的相对表达量,间接免疫荧光法测定热应激处理1 h后HSP70在鸡原代心肌细胞中的定位。结果表明:热应激后HSP70在鸡原代心肌细胞中的表达量呈先上升后下降趋势;与对照相比,热应激8 h后鸡原代心肌细胞的HSP70表达量显著降低(P0.05);HSP70在对照与热应激1小时时的心肌细胞中均有表达,且在细胞浆与细胞核中均有分布,但主要分布于细胞浆中。说明热应激对HSP70在鸡原代细胞中的表达产生影响,但对其在细胞中的分布无明显影响。     

热休克蛋白70的生物学功能及其诱导剂——谷氨酰胺在畜牧业中的应用

<正>热休克蛋白(heat shock protein,HSP)或热应激蛋白(heat stress protein,HSP)是机体受到应激原的刺激后产生的几族高度保守的蛋白质,对维持细胞生存和内环境的稳定起重要作用。其中热休克蛋白70(HSP70)是最重要的一种HSP,它具有多种生物学功能,包括分子伴侣功能、参与免疫反应、抗细胞凋亡功     

急性热应激对鸡呼吸系统损伤及肺脏热休克蛋白表达的影响

为探讨热应激对鸡肺脏组织损伤的影响,将60只35日龄SPF鸡随机分为对照组,热应激1、2、3、5、10 h组,每组10只,试验开始后环境温度迅速从25℃升高到35℃,观察热应激组鸡临床症状,热应激结束迅速剖杀、取病料,检测血清pH值、乳酸脱氢酶(LDH)、钾离子和钙离子浓度,石蜡切片检测肺脏组织结构,Western blot检测肺脏组织中热休克蛋白(HSPs)表达量。结果显示:与对照组相比,热应激组血清pH值显著升高(P<0.05),LDH水平均极显著升高(P<0.01),随着热应激时间增加,血钾和血钙浓度开始降低,热应激5、10 h后血钾和血钙浓度显著减低(P<0.05);病理组织学结果显示热应激后,肺组织内血管充血,肺房结构基本完整,热应激5 h后肺房内有大量的红细胞存在,热应激10 h肺房内的异物减少,但肺上皮细胞大量脱落,组织结构损伤严重;Western blot结果显示与对照组相比,HSP27和HSP72表达量极显著升高(P<0.01)HSP60表达量在热应激1 h后显著升高(P<0.05),随后呈降低的趋势,HSP90表达量在热应激5 h后显著升高(P<0.05),随后呈降低的趋势,HSC70表达量无明显变化。热应激可对鸡呼吸系统造成损伤,提高肺脏HSPs表达量。     

Induction of heat-shock proteins and phagocytic function of chicken macrophage following in vitro heat exposure.

The protein profiles and phagocytic ability of Sephadex-elicited chicken peritoneal macrophages were examined following heat-shock exposure. Macrophage cultures were exposed to various temperatures, time exposures and recovery periods. Densitometric analysis of SDS-PAGE autoradiographs revealed that heat-induced macrophages synthesized three major (23, 70 and 90 kD) heat-shock proteins (HSPs). The optimal temperature and time for induction of these HSPs was 45-46 degrees C for 1 h, with a variable recovery period for each HSP. Macrophages exposed to 45 degrees C for 30 and 60 min were significantly depressed in phagocytosis of uncoated sheep erythrocytes (SE) under 45 degrees C incubation conditions. However, phagocytosis of antibody-coated SE was not affected when compared to 41 degrees C control cultures. Macrophages allowed to recover at 41 degrees C following heat-shock exhibited no alterations in their phagocytic ability for either antibody-coated or uncoated SE. This study suggests that heat shock induces three major HSPs in chicken peritoneal macrophages in addition to maintaining their Fc-mediated phagocytic function while significantly depressing their nonspecific phagocytosis.     

Dietary supplementation of Zingiber officinale and Zingiber zerumbet to heat‐stressed broiler chickens and its effect on heat shock protein 70 expression,blood parameters and body temperature

The present study was conducted to assess the effects of dietary supplementation of Zingiber officinale and Zingiber zerumbet and to heat‐stressed broiler chickens on heat shock protein (HSP) 70 density, plasma corticosterone concentration (CORT), heterophil to lymphocyte ratio (HLR) and body temperature. Beginning from day 28, chicks were divided into five dietary groups: (i) basal diet (control), (ii) basal diet +1%Z. zerumbet powder (ZZ1%), (iii) basal diet +2%Z. zerumbet powder (ZZ2%), (iv) basal diet +1%Z. officinale powder (ZO1%) and (v) basal diet +2%Z. officinale powder (ZO2%). From day 35–42, heat stress was induced by exposing birds to 38 ± 1 °C and 80% RH for 2 h/day. Irrespective of diet, heat challenge elevated HSP70 expression, CORT and HLR on day 42. On day 42, following heat challenge, the ZZ1% birds showed lower body temperatures than those of control, ZO1% and ZO2%. Neither CORT nor HLR was significantly affected by diet. The ZO2% and ZZ2% diets enhanced HSP70 expression when compared to the control groups. We concluded that dietary supplementation of Z. officinale and Z. zerumbet powder may induce HSP70 reaction in broiler chickens exposed to heat stress.     

热应激对家禽HPG轴结构和繁殖功能的影响及HSPs的作用

高温环境容易引起家禽热应激,造成家禽下丘脑-垂体-性腺(hypothalamic-pituitary-gonadal,HPG)轴各组织结构不同程度的损伤,进而影响家禽正常繁殖功能,给集约化饲养管理模式下家禽养殖业造成一定的损失。热休克蛋白(HSPs)是一类保护性蛋白,它能在家禽遭受高温应激时发挥分子伴侣作用,一定程度上保护机体免受高温刺激的损伤。文章对热应激条件下家禽的HPG轴各组织结构、繁殖功能的影响及HSPs的作用进行综述,为家禽生产提供相关资料。     

小鼠卵巢HSP70诱导表达的免疫荧光组织化学研究

试验采用免疫荧光组织化学法,检验了经历不同温度热应激后,在不同恢复期内小鼠卵巢组织HSP70的诱导表达,确立了HSP70诱导表达模式。结果表明：随着热应激温度的升高,卵巢组织HSP70的表达量增加,表达持续期延长,特别是经历了预应激小鼠的HSP70表达更显著。这说明在一定的热应激条件下。小鼠卵巢组织可发生HSP70的诱导表达,其表达量和表达持续期与热应激强度相关,而且热应激可显著促进HSP70的表达。     

夏季奶牛血液中热休克蛋白的表达量

选择40头体质健康的中国荷斯坦奶牛,根据胎次和泌乳天数,按照随机区组试验设计分为Ⅰ组（30kg/d）、Ⅱ组（30～35kg/d）、Ⅲ组（35～40kg/d）和Ⅳ组（40kg/d）。热应激前、热应激前期、热应激中期、热应激后期和热应激后分别于尾静脉采血,用ELISA试剂盒测定热休克蛋白（HSP）27,70,90的表达量。结果显示,Ⅳ组HSP27表达量最高,Ⅱ组表达量最低,Ⅳ组、Ⅲ组和Ⅰ组均显著高于Ⅱ组（P〈0.05）。HSP70表达量各组间没有明显差异,但随产奶量呈线性增加（P〈0.05）。HSP90的表达量,Ⅳ组和Ⅲ组明显高于Ⅱ组（P〈0.05）。HSP27的表达量热应激后差异较大;HSP70的表达量各组整个过程差异较大;HSP90的表达量在热应激前、热应激前期和热应激后差异较大。总之,在热应激过程,高产奶牛血清中热应激蛋白的表达量较高,HSP70表达量随产奶量呈线性增加,而不同热应激蛋白的变化规律差异较大。     

Effect of heat challenge on peripheral blood mononuclear cell viability: comparison of a tropical and temperate pig breed

We evaluated the effect of heat challenge on cell viability, concanavalin A-induced proliferation and heat shock protein (HSPs) mRNA expression in peripheral mononuclear blood cells (PBMC) isolated from Creole (CR) and Large White (LW) pigs. The PBMCs were cultured for 9 h at 37°C before being subjected to heat challenge: (1) at 42°C or 45°C for 2, 4, 6 and 9 h to monitor cell viability;(2) at 45°C for 2 and 9 h followed by stimulation for 24 h at 37°C with concanavalin A to evaluate mitogen-induced proliferation; and (3) at 45°C for 3, 6 and 9 h to measure induction of HSP70.2 and HSP90 mRNA. Cell viability was affected by breed and temperature (P < 0.01), and the viability decrease caused by heat challenge was greater for LW than CR pigs. For mitogen-stimulated PBMCs, incubation at 45°C reduced lymphoblastogenesis equally in both breeds (P < 0.01). Although heat challenge for 3 and 6 h at 45°C induced expression of HSP70.2 and HSP90 mRNA, no breed difference was observed. In conclusion, differences in heat resistance between these two breeds at the whole organism level are reflected at the cellular level. Neither HSP70.2 nor HSP90 mRNA expression levels explain this effect.     

Comparative studies on temperature threshold for heat shock protein 70 induction in young and adult Murrah buffaloes

To know the temperature threshold for heat shock protein 70 (HSP70) induction in lymphocytes and to assess physiological changes, if any, in relation to HSP70 induction in young and adult Murrah buffaloes, this study was divided into two parts: I. In vivo study: where assay of HSP70 was performed in blood samples collected from acutely exposed young and adult Murrah buffaloes (n = 6) inside a climatic chamber at 40, 42 and 45 °C for 4 h and thermoneutral temperature (22 °C). Physiological parameters viz., rectal temperature, respiratory rate, pulse rate and skin temperature of different body parts were monitored to assess magnitude of stress in the animals owing to thermal exposure II. For in vitro study, equal numbers of lymphocyte cells were separated from blood collected from young and adult buffaloes and were subjected to four temperature treatments (38, 40, 42 and 45 °C) for 4 h. A significant increase (p < 0.05) in all the physiological parameters in both young and adult buffaloes was observed after exposure to 40, 42 and 45 °C for 4 h as compared to 38 °C. The average plasma HSP70 concentrations (ng/ml) were significantly higher (p < 0.05) at 40, 42 and 45 °C as compared to 38 °C in both young and adult and were higher in young than adult buffaloes at 38 and 45 °C. Heat shock protein 70 level in lymphocyte lysate showed highest concentration after 3-h exposure to all temperatures (40, 42 and 45 °C) in both young and adult buffaloes. The intensity of changes of all physiological parameters was more in young animals than in the adults indicating the greater susceptibility of younger animals to heat stress and was found to be changed at around 40 °C when animals were exposed to different temperatures, indicating the possibility that HSP70 production may be initiated at this temperature which is 2 or 3 °C higher than core body temperature.     

热应激对小鼠附植前早期胚胎HSP70的诱导表达

选择超数排卵后怀孕昆明小鼠75只,以注射人绒毛膜促性腺激素(hCG)后的胚胎发育阶段为标准,分别在合子、2细胞、4细胞、8~16细胞和囊胚期对孕鼠进行了37℃、39℃、41℃和43℃热应激4 h。热应激后取胚,用免疫荧光细胞化学法检验胚胎热休克蛋白70(HSP70)的诱导表达。结果表明,HSP70在4细胞以上胚胎内呈阳性表达,相对高温时其表达量增加,囊胚时最显著。可见,附植前早期胚胎HSP70的热诱导表达与细胞期和温度密切相关。