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1.
HSP70在急性热应激蛋鸭脾脏表达变化的研究   总被引:1,自引:0,他引:1  
采用半定量RT-PCR与Western-blotting方法检测相同温度、不同时间(3、5、7h)热应激处理鸭脾脏组织中HSP70基因的表达水平,以研究热应激对HSP70基因表达的影响。结果表明:与对照组相比,热应激组蛋鸭脾脏HSP70 mRNA转录水平在热应激进行到3h时呈现下降趋势,但是差异不显著;而进行到5h时则出现显著(P0.05)的回升;热应激组蛋鸭脾脏HSP70蛋白表达水平在热应激进行到5h时出现显著下降(P0.05),表明急性热应激对蛋鸭脾脏组织HSP70基因表达具有重要影响。  相似文献   

2.
采用束缚浸水法建立大鼠应激性溃疡模型,探讨了胃泌素、胰高血糖素、血栓纱B2(TXB2)的变化与与应激性溃疡相关性及应激宁的抑制效应。结果表明,应激组血浆、胃组织的3处激素水平的均极显著的高于对照组,应激+应激宁组与应激组相比较,3处激素的水平极显著或显著降低,或降低不显著,提示这3种激素可能从不同角度参与应激性溃疡的形成,同时,应激宁对应激性溃疡的形成具有抑制作用。  相似文献   

3.
本试验以SD大鼠为试验动物,利用摇床模拟公路运输过程中的摇晃、高温、噪音、饥渴、拥挤、碰撞6个主要的刺激因子,构建了大鼠运输应激模型.应激条件为60 r/min,35℃,每天应激2h.体重为270±20 g,20只SD大鼠被随机均分为4组:对照组、应激1d组、应激2d组、应激3d组.在应激结束后,应激组大鼠体温均显著升高(P<0.05),体重显著下降(P<0.05),血清中皮质醇含量显著升高(P<0.05).试验结果与实际公路运输结果一致,表明运输应激模型构建成立.形态学研究显示,运输应激造成了大鼠空肠组织损伤,与对照组相比从应激第1天到应激第3天空肠绒毛脱落逐渐加剧.荧光定量PCR研究结果则显示,Hsp27,Hsp70和Hsp90 mRNA的表达量也剧烈的升高.本试验研究表明,随着应激时间增加,大鼠空肠损伤逐渐加重,且Hsps mRNA表达量升高,表明Hsps的表达量与运输应激造成空肠的损伤严重程度有关.运输应激后表达量急剧升高的保护性分子Hsps,是动物抵抗运输应激的重要机制之一.  相似文献   

4.
通过分析冷刺激对大鼠血清中细胞因子、皮质酮(CORT)、促肾上腺皮质激素(ACTH)的含量以及外周血淋巴细胞中热休克蛋白70(Hsp70)表达量的影响,构建大鼠急性冷应激模型,旨在为其相关研究提供研究基础。本试验急性冷刺激时间为12h。利用ELISA方法检测各组大鼠血清中IL-2、IL-4、IL-6、IL-10、TNF-α、CORT、ACTH的含量;采用Western blot和qRT-PCR方法检测各组大鼠外周血淋巴细胞中HSP70及mRNA表达量。结果表明,与对照组相比,急性冷刺激组大鼠血清中IL-2和ACTH的含量显著升高(P0.05),IL-4、IL-6、TNF-α和CORT的含量极显著升高(P0.01),IL-10无显著变化(P0.05);外周血淋巴细胞内HSP70及其mRNA表达量呈显著升高水平(P0.05)。成功构建大鼠急性冷应激模型。  相似文献   

5.
本试验通过检测大鼠肠道中热休克蛋白基因的表达,探究不同剂量香苇二石口服液对热应激的预防和治疗作用。45只健康雄性大鼠随机分为对照组、热应激组、自然恢复组、高剂量预防组、中剂量预防组、低剂量预防组、高剂量治疗组、中剂量治疗组、低剂量治疗组,建立高温应激模型,利用Real time-PCR技术检测大鼠小肠中HSP27、HSP70、HSP90 m RNA的表达变化。结果表明,高温刺激能极显著增加HSPs m RNA的表达量(P<0.01);香苇二石口服液能显著(P<0.05)和极显著(P<0.01)抑制HSPs m RNA的高表达。表明香苇二石口服液高剂量有极显著预防作用,中剂量有极显著治疗作用。  相似文献   

6.
急性冷、热应激对肉鸡肝脏和心脏组织中HSPs表达的影响   总被引:2,自引:0,他引:2  
将45只28日龄罗斯308肉鸡随机均分为三组进行急性应激处理:热应激组在急性热应激下持续饲养6 h(温度为39℃,湿度为90%);冷应激组在急性冷应激刺激下持续饲养6 h(温度为11℃,湿度为90%);对照组鸡群在正常饲养条件下饲养(温度为26℃,湿度为60%)。采用实时荧光定量PCR(q RT-PCR)分别对肝脏和心脏组织中的4种热应激蛋白基因(HSP60、HSP70、HSP90、HSP110)的表达量进行对比分析。结果表明:在心脏组织中,HSP60表达量在冷、热应激下均无显著变化(P0.05);HSP90表达在冷应激下显著下降(P0.05);HSP70表达在冷、热应激下均显著上升(P0.05、P0.01);HSP110表达在冷应激下无显著变化,在热应激下极显著上升(P0.01)。在肝脏组织中,HSP60在冷应激下表达量显著下降(P0.05),热应激下表达无显著变化(P0.05);HSP70和HSP110在热应激条件下表达量极显著上升(P0.01);HSP90表达量在冷、热应激下均无显著变化(P0.05)。结果揭示:急性热应激相比于急性冷应激对应激蛋白的表达影响更显著;在急性热应激条件下,心脏和肝脏组织中HSP70和HSP110表达量均极显著上升。  相似文献   

7.
选取15头体况相似的健康肉牛作为研究对象,随机分为运输0h组、运输3h组和运输6h组,每组5头肉牛,采用公路运输方式运输(平均速度为80km/h)。采集各组牛肝脏和脾脏组织样本,用酶联免疫吸附试验(ELISA)和荧光定量PCR(qRT-PCR)2种试验方法检测各组肝脏和脾脏组织中应激蛋白-热休克蛋白70(HSP70)和急性期蛋白-C反应蛋白(CRP)含量的变化。qRT-PCR检测结果显示:与运输0h组相比,运输3h和6h肝脏组织中HSP70mRNA的表达量极显著升高(P0.01);而脾脏组织中HSP70mRNA只在运输应激6h后表达量显著增加(P0.05)。与0h组相比,运输应激后肝脏组织中CRP mRNA的表达量极显著升高(P0.01),脾脏组织中CRP mRNA的表达量显著升高(P0.05)。ELISA检测结果显示:与0h组相比,运输应激3h和6h肝脏中的HSP70质量浓度极显著升高(P0.01),脾脏中HSP70的质量浓度在运输6h后显著升高(P0.05);肝脏和脾脏CRP的表达在运输应激6h后极显著升高(P0.01)。结果表明:不同运输时间的运输应激可导致肝脏和脾脏中HSP70和CRP的浓度和mRNA转录水平升高,且肝脏中HSP70和CRP的表达量高于脾脏。HSP70和CRP可作为肉牛运输应激候选诊断标志物。  相似文献   

8.
为了研究复方中药对热应激肉鸡肝脏HSP70mRNA表达的调控效果;试验选取120只18日龄AA+肉仔鸡,随机分为4组,即复方中药组、维生素组、纯中药组和高温对照组进行试验,试验后每组迅速宰杀3只试验鸡,取肝脏组织进行HSP70mRNA表达的检测。结果表明:高温应激后,复方中药冲剂组和纯中药冲剂组鸡肝脏中HSP70 mRNA表达显著增加,分别比高温对照组、维生素组高2.98倍、3.28倍和4.66倍、5.13倍,说明本试验复方中药可通过上调HSP70 mRNA表达,提高机体热耐受水平,达到保护细胞作用。  相似文献   

9.
研究蛋鸭热应激条件下不同组织中热休克蛋白70(HSP70)基因mRNA表达规律,为揭示蛋鸭热应激反应机理提供参考。60只绍兴蛋鸭25℃饲养20d,其中30只进行40℃热应激处理1h,采取荧光定量PCR方法分别测定不同组织中HSP70mRNA的表达水平。试验组与对照组相比,肝脏、脾脏和胰腺组织HSP70mRNA表达差异不显著,其他6种组织HSP70基因表达量均有不同程度的增加,试验组心脏、腿肌、垂体、下丘脑、胸肌、肾脏中HSP70基因表达量分别高于对照组3.6倍、2.8倍、2.7倍、2.4倍、2.2倍和1.57倍。  相似文献   

10.
选取1日龄健康雁鹅60只,随机分为6组,单笼饲养于环控仓内。6周龄时,A组置于(22±1)℃作为对照,B、C、D、E、F组分别施于2、4、6、8 h和10 h(40±1)℃的急性热应激处理,处理后立即剖杀,取心、肝、肺和肾脏组织,实时荧光定量PCR测定HSP70 mRNA表达量,免疫组织化学方法测定HSP70表达量。结果表明:急性热应激处理可显著提高各组织中HSP70 mRNA及HSP70蛋白的表达水平,但不同组织出现显著变化的时间以及随热应激时间延长的变化规律存在差异。心脏组织HSP70 mRNA和蛋白表达量均在热应激2 h后出现显著升高(P0.05),热应激4 h后达到最高值,8 h后降至对照组水平,遵循二次曲线回归模式;肝脏热应激4~10 h间HSP70及其mRNA均显著高于对照组(P0.05),但未表现出显著的线性与二次回归趋势;肺脏HSP70 mRNA表达水平在热应激2~10 h间显著高于对照组(P0.05),HSP70则从热应激4 h起才显著升高,但二者随热应激时间延长均呈显著的线性升高规律;与肺相似,肾脏HSP70及其mRNA表达量随热应激时间的变化趋势也遵循线性回归模式,两者均在热应激4~10 h显著升高(P0.05)。  相似文献   

11.
The aim of the present study was to investigate the pathological injury and the expression of heat shock proteins in the caprine lung, trachea and bronchus under transport stress. 12 healthy male goats were selected and randomly divided into three groups. The control group (non-transported group), 2 hr transport-treated group and 6 hr transport-treated group. Morphological changes as well as the expression of heat shock proteins (HSPs, mainly HSP27, HSP70 and HSP90) in three parts of the respiratory tract were examined. Our results showed swollen mucosa and congestive blood vessels in mucous layer and submucosa, inflammatory cell infiltration as well as degeneration and necrosis of mucosal epithelial cells in trachea and bronchus of the transport-treated groups. The epithelial cells were degenerated, and the exfoliated cells and debris could be seen in the alveolar cavity. The results of immunohistochemistry showed that HSP27 and HSP70 were strongly expressed in tracheal and bronchial epithelium, glandular epithelium, vascular endothelium and bronchiole epithelium. And the amount of positive inflammatory cells was increased in transport-treated groups. Western blot results indicated that the expression of all three proteins had no obvious difference among the three groups in bronchi (> .05). In trachea, there was no significant difference in the expression of heat shock proteins among the three groups except that the expression of HSP70 which was obviously higher in the two transported groups than the control group (p < .05). The expression level of HSP70 in the 2 hr transport-treated group was significantly higher than the 6 hr group (p < .05) and control groups (p < .05). However, there was no significant difference in the expression level of HSP27 and HSP90 in three groups (> .05). In conclusion, our data showed that transport stress could damage the caprine respiratory system.  相似文献   

12.
本试验以谷氨酰胺作为热休克蛋白70(HSP70)诱导剂,从细胞凋亡线粒体信号转导途径探讨HSP70对热应激猪小肠上皮细胞(IPEC-J2)凋亡相关因子的影响。试验以体外培养的猪小肠上皮细胞为模型,分别加入不同浓度(1、2、4、6、8、10 mmol/L)谷氨酰胺的培养基中培养24 h,提取总RNA和总蛋白,用实时荧光定量PCR和Western blot检测HSP70 mRNA和蛋白的相对表达量,筛选出谷氨酰胺诱导HSP70表达的最佳浓度。取对数生长期的细胞分为3组,分别为对照组(Con组,37℃培养12 h)、热应激组(Hs组,42℃培养12 h)、谷氨酰胺组(Gln组,6 mmol/L谷氨酰胺、42℃培养12 h)。采用Annexin V/PI双染法检测细胞凋亡水平,实时荧光定量PCR和Western blot分别检测HSP70、凋亡酶激活因子-1(Apaf-1)、半胱氨酸天冬氨酸蛋白水解酶-9(Caspase-9)、半胱氨酸天冬氨酸蛋白水解酶-3(Caspase-3)、B淋巴细胞瘤-2(Bcl-2)的mRNA和蛋白的相对表达量。结果表明:谷氨酰胺对HSP70上调表达呈先上升后下降的趋势,6 mmol/L的谷氨酰胺诱导时HSP70 mRNA和蛋白相对表达量最高,显著高于其他浓度(P<0.05),作为后续试验中Gln组的诱导条件。与Con组相比,Hs组细胞凋亡率极显著增加(P<0.01),HSP70、Apaf-1、Caspase-3、Caspase-9 mRNA和蛋白的相对表达量极显著升高(P<0.01),而Bcl-2 mRNA和蛋白的相对表达量极显著降低(P<0.01)。与Hs组相比,Gln组细胞凋亡率极显著降低(P<0.01),但仍极显著高于Con组(P<0.01);Gln组HSP70 mRNA和蛋白的相对表达量极显著升高(P<0.01);Gln组Apaf-1、Caspase-3、Caspase-9 mRNA和蛋白的相对表达量极显著降低(P<0.01),但极显著高于Con组(P<0.01);Gln组Bcl-2 mRNA和蛋白的相对表达量极显著升高(P<0.01),但极显著低于Con组(P<0.01)。由此可见,上调HSP70的表达可有效缓解热应激导致的猪小肠上皮细胞凋亡。  相似文献   

13.
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。  相似文献   

14.
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。  相似文献   

15.
以东北野猪成纤维细胞为研究对象,利用实时荧光定量PCR技术,分析4、15、25和32℃不同温度冷应激条件下细胞内HSPT0mRNA表达变化规律。结果显示在冷应激的低温处理过程中,HSP70mRNA转录量在各温度均为见显著增加;在冷应激的复温培养过程中,HSP70mRNA转录量在4℃和15℃处理后4h后,复温培养的4~8h显著增加(P〈O.05);在25℃和32℃处理后4h后,复温培养未能诱导细胞内HSP70mRNA转录量显著增加(P〉0.05);细胞在4℃和15℃处理2、4、6和8h后,复温培养4h,HSP70mRNA转录量随处理温度的减低和时间的延长而增加,处理4~8h组显著高于对照组(P〈0.05),在25℃和32℃处理2、4、6和8h后,复温培养未能显著诱导HSP70mRNA的转录(P〉0.05)。结果表明,冷应激诱导东北野猪成纤维细胞内HSP70mRNA转录量的增加,不是发生在低温处理的应激阶段,而是发生在复温后的细胞应激阶段,温和冷应激(25~32℃)未能诱导复温后HSP70mRNA转录量的显著增加,强度冷应激(4~15℃)诱导复温后HSP70mRNA转录量的显著增加,且与冷应激的强度和时间成正比。  相似文献   

16.
To contribute to early diagnosis and treatment of gastric ulcer of foals, we examined the gastric mucosa of healthy and affected foals using an endoscope. In healthy foals, the characteristic changes in the development of the squamous mucosa were seen mainly in the squamous mucosa, and maturation of the squamous mucosa in the greater curvature (GC-S) occurred more slowly than that of the squamous mucosa in the lesser curvature (LC-S). Epithelial desquamation in the LC-S and GC-S was observed between 6 and 90 days but was not observed in the LC-S at about 60 days, whereas it was observed in the GC-S until 90 days old. These findings suggest that there is a difference in the development of the gastric mucosa by region and that desquamation continues over a term longer than studies have reported in the past. In the affected foals, the minimum age at which gastric ulcer was observed was 4 days old. Gastric ulcers formed predominantly in the squamous mucosa (LC-S and GC-S) of foals with an immature mucosa before the weaning period, and the peak incidence occurred between 61 and 90 days old. The differences in the ulceration sites were considered to depend on the difference in the development (maturation) stage of the squamous mucosa. The grading score of the gastric ulcer increased with the growth of the affected foals. The gastric ulcer might be enhanced greatly by stress in the weaning period.  相似文献   

17.
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。  相似文献   

18.
为探讨金属硫蛋白(MT)对热应激奶牛血淋巴细胞凋亡基因和HSP70基因表达的影响及其机理,选取25头中国荷斯坦奶牛经产泌乳母牛,随机均分为A、B、C、D、E 5组,分别按每头0(对照),16.0,24.0,32.0和40.0 mg颈静脉注射Zn-MT,于试验第1(注射Zn-MT之前),7和14天逐头采取血样,测定不同剂量的外源性MT对热应激奶牛血淋巴细胞HSP70、Bcl-2、Baxp53基因表达水平的影响。结果表明,补给外源性MT后,4个试验组奶牛的产奶性能都有不同程度提高,其中B组奶牛的产奶量比A、C、D和E组分别高出20.94%(P<0.05),15.83%(P>0.05),10.94%(P>0.05)和8.85%(P>0.05);各试验组HSP70基因的表达水平都有所上升,其中D组比对照组高出130.00%,达到显著(P<0.05)水平;各试验组Bcl-2基因的表达水平均高于对照组,其中C组达到显著(P<0.05)水平;B、D组Bax基因的表达水平和C、E组p53基因的表达水平较对照组分别降低了22.00%,13.00%和22.00%,34.00%,但均未达到显著(P>0.05)水平。说明外源性MT能改善热应激奶牛的产奶性能并有效调控奶牛血淋巴细胞HSP70、Bcl-2、Baxp53基因的表达水平,且表现出较为明显的剂量效应。  相似文献   

19.
Thoroughbred and Standardbred horses in race training were selected at random and examined for gastric ulcers. The mucosa of the stomach was examined by endoscopy and was scored for ulcers on a scale of 0 (no ulcers, normal mucosa) to 3 (severe ulceration with at least five deep lesions). A total of 79 horses were examined; 37 of these horses had one to four follow-up examinations, and 42 horses had only an initial gastroscopic examination. On the first examination, 33 of 79 (42%) horses had normal mucosa (ulcer score = 0), and 22 of 79 (28%) had ulcers with an ulcer score of 2 or greater. Seven horses (9%) had severe gastric ulceration (ulcer score = 3). The average ulcer score for all 79 horses on initial examination was 1.1 and 1.2 for the 37 horses with follow-up gastroscopic examination. Males (including male castrates) had slightly higher ulcer scores (1.2) than females (1.0), and 2-year-old horses had lower ulcer scores (0.7) than horses older than 3 years (1.4), but these differences did not reach statistical significance. Each of these average scores was significantly (P < .05) greater than zero. For the 37 horses with follow-up examinations, the males had higher scores (1.6) than females (0.7), and 2-year-old horses had higher ulcer scores (1.0) than 3-year-old horses (0.7) and lower ulcer scores than horses older than 3 (1.8), which were statistically significant. Follow-up examinations revealed ulcer scores ranging from 0 to 3; 23 of 37 (63%) horses had an ulcer score of 2 or greater, and 4 of 37 (12%) horses had severe ulcer disease. Average ulcer scores were significantly higher at subsequent examinations than at the initial examination. Risk of gastric ulceration increases with the length of time horses are in race training.  相似文献   

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