Decreased serum heat shock protein 60 levels in newly diagnosed immune thrombocytopenia patients

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by peripheral thrombocyte destruction. In some autoimmune disorders, heat-shock proteins (HSP) are suggested to be an important antigenic factor. In this study, we demonstrated the serum free levels of HSP60, HSP70, anti-HSP60, and anti-HSP70 in ITP patients and healthy controls. Twenty-eight newly diagnosed ITP patients, 35 ITP patients in chronic phase, and 25 healthy controls were enrolled to this study. Serum levels of HSP60, HSP70, anti-HSP60, and anti-HSP70 were determined by the ELISA method. Serum HSP60 levels of newly diagnosed ITP patients were significantly decreased when compared with both chronic phase ITP patients and healthy controls. HSP60 levels of ITP patients (both newly diagnosed and chronic phase) with thrombocyte counts more than 30?×?10⁹/L were significantly increased compared with ITP patients with thrombocyte counts less than 30?×?10⁹/L and there was a positive correlation between thrombocyte counts and serum free HSP60 levels in ITP patients. This is the first study demonstrating the extracellular HSP levels in adult ITP patients. HSPs are shown to have a place in the pathogenesis of many autoimmune disorders. Low level of HSP60 may lead to lack of anti-inflammatory response due to less Treg activation, hence, could be a counterpart in the pathogenesis of ITP. Further studies are needed to understand the role of HSPs in the pathogenesis of ITP and whether they can be used for diagnosis, prognosis, and treatment of ITP.     

可溶性热休克蛋白60与动脉粥样硬化的相关性及药物干预的实验研究

目的:探讨可溶性热休克蛋白60(sHSP60)与实验大鼠主动脉粥样硬化的关系及药物对其影响。方法:建立大鼠动脉粥样硬化模型,随机分为对照组、高脂组、血脂康组及阿司匹林组,并对其血脂水平、血清sHSP60进行检测,应用苏木精-伊红及油红O染色观察各组血管形态学变化。结果:血脂康组血脂水平较高脂组及阿司匹林组明显降低,阿司匹林组sHSP60水平较高脂组及血脂康组明显下降;血脂康组和阿司匹林组血管脂质沉积较高脂组减少。结论:sHSP60可能是与血脂水平无关的动脉粥样硬化预测因子;血脂康、阿司匹林均可有效减少血管壁脂质沉积。     

HSP70,HSP90在结肠癌中表达及其和生物学行为的相关性

目的:探讨HSP70,HSP90在人结肠癌中的表达情况及其和结肠癌细胞生物学行为的相关性.方法:结肠癌患者手术标本40例,分别以癌组织、癌旁2cm组织和远离癌灶的正常黏膜组织作为病例组和对照组.用亲和免疫组织化学技术方法检测HSP70和HSP90,并收集该40例患者的临床Duke's分期,分析HSP70和HSP90蛋白的阳性程度和结肠癌临床分期是否存在相关性.结果:癌组织HSP70和HSP90的表达明显增高,癌组织、癌旁组织和正常黏膜组织三组之间HSP70和HSP90表达率有明显差异(HSP70:82.5%vs52.5%vs25%,χ2=26.58,P=0.000;72.5%vs42.5%vs22.5%,χ2=20.41,P=0.000);HSP70和HSP90蛋白的阳性程度和结肠癌临床分期存在相关性(HSP70:tau_b=0.392,P=0.006;HSP90:tau_b=0.396,P=0.006).结论:结肠癌细胞存在HSP70和HSP90蛋白的过度表达,HSP70,HSP90和结肠癌细胞的生物学行为存在相关性.     

Induction and intracellular localization of a 72-kDa heat shock protein in rat gastric mucosa after water-immersion stress

We investigated the expression and changes in the intracellular localization of a 72-kDa heat shock protein (HSP72) in rat gastric pyloric and fundic mucosa before and after water-immersion stress. Severe mucosal damage was found in the fundic mucosal area of the stomach after this stress. However, no mucosal lesion developed in the pyloric mucosal area. HSP72 in both the soluble and insoluble fractions of the pyloric and the fundic mucosal areas was significantly increased after water-immersion stress, peaking 6 h after the initiation of the stress. The increase in HSP72 was more significant in the pyloric mucosal area than in the fundic mucosal area under both normal and stress conditions. The increase of HSP72 in the pyloric mucosal cells occurred prior to the formation of the mucosal lesions, whereas the increase of HSP72 in the fundic mucosal cells was observed after ulcer formation. An immunohistochemical study showed that HSP72 was constitutively expressed in the cytoplasm of the gastric mucosal cells, and that the intranuclear induction of HSP72 was remarkably intense in the pyloric mucosal cells, especially in the proliferative zone, compared with the fundic mucosal cells. Our results may suggest that HSP72 has an important cytoprotective function in gastric mucosal cells and that there is a “biophysical” difference between pyloric and the fundic mucosal cells.     

Correlation between clinicopathology and expression of heat shock protein 70 and glucose-regulated protein 94 in human colonic adenocarcinoma

AIM: To investigate the correlation between clinicopathology and expression of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human colonic carcinoma. METHODS: The expression of HSP70 and grp94 was studied in 80 human colonic cancers with or without metastasis as well as in their adjacent mucous membrane by way of immunohistochemistry and pathology photograph analysis. RESULTS: The expression of HSP70 and grp94 was significantly higher in cancer than that in adjacent mucous membrane (92.5%, 85.0% vs 56.3%, 42.5%, P<0.01). HSP70 and grp94 expressed higher in moderately- and poorly-differentiated colonic cancers than that in their adjacent tissues (93.7%, 87.5%; 100%, 90% vs56.3%, 42.5%;P<0.01). Dukes C and D stages of colonic cancers showed higher positive rates than Dukes A and B stage groups (97.1%, 91.2%; 100%, 90.9%; vs 80%, 70%; 78.6%, 71.4%; P<0.05). There were definite differences in HSP70 and grp94 expression between metastasis groups and non-metastasis groups (100% vs 75%, 100% CONCLUSION: The HSP70 and grp94 expression rates in colonic cancer groups are significantly higher than that in their adjacent mucous membrane. The HSP70 and grp94 expression in poorly-differentiated colonic cancers with metastasis is significantly higher than well-differentiated cancers without metastasis. The overexpression of HSP70 and grp94 can be used as diagnostic or prognostic markers for colonic cancer.     

肠化胃黏膜病变组织HSP60的表达与幽门螺杆菌感染的相关性

目的:探讨HSP60在肠化胃黏膜组织中的表达及与H pylori感染之间的关系.方法:对171例肠化胃黏膜组织其中I型肠化54例、Ⅱ型肠化76例、Ⅲ型肠化41例,对照组浅表性胃炎40例,胃癌49例,采用SP免疫组织化学技术进行胃黏膜HSP60的检测,用ELISA,H pylori IgG抗体检测及H pylori-DNA PCR方法进行H pylori感染情况的判定.结果:HSP60在肠化胃黏膜组织中的表达主要为中等强度阳性,Ⅰ,Ⅱ,Ⅲ型肠化胃黏膜组织的阳性率分别为35.19%,51.32%,75.61%,过表达率分别为1.9%,3.9%和19.5%;浅表性胃炎组织中的表达多为弱阳性或中等强度阳性,阳性表达率为30.00%,与浅表性胃炎相比肠化胃黏膜组织中HSP60的表达显著增加(P＜0.01);HSP60在胃癌组织的表达以强阳性和中等强度阳性为主,阳性表达率为73.47%,过表达率为34.69%,明显高于Ⅰ,Ⅱ型肠化胃黏膜组织中的表达,与Ⅲ型肠化胃黏膜组织相比差异不显著(P＞0.05),肠化胃黏膜组织的Hpylori感染和HSP60表达有明显的相关性(r=-0.191,P=0.012)结论:HSP60的过度表达发生在胃黏膜病变的进展过程中;肠化胃黏膜组织中H pylori感染和HSP60表达有明显的相关性.     

Impaired humoral immune response against mycobacterial 65-kDa heat shock protein (HSP65) in patients with inflammatory bowel disease

Since only scarce data are available on the immune response against heat shock proteins (HSP) in inflammatory bowel disease (IBD), we have measured with an ELISA method serum levels of IgG, IgA, and IgM antibodies to mycobacterial HSP65 and human HSP60 in 66 patients with Crohn's disease (CD), 42 patients with ulcerative colitis (UC), and 126 age- and gender-matched healthy controls. Serum concentration [median (25th–75th percentiles) of IgG anti-HSP65 antibodies was substantially lower in patients with either CD (P < 0.01) or UC (P < 0.001) than in healthy controls, while no difference was found in the levels of anti-HSP60 antibodies. Low anti-HSP65 antibody levels were measured in patients with active CD and in both active and inactive UC, and only in IBD patients with no extraintestinal manifestations. In conclusion, our present findings indicate that an abnormal immune response to bacterial HSP65 or some epitopes of the protein may contribute to the dysregulation of host defenses against certain intestinal bacteria.     

热休克蛋白60及重组热休克蛋白60对U937细胞IL-6分泌量的影响

目的观察重组热休克蛋白60（hsp60）对U937细胞分泌细胞因子IL-6的影响。方法对幽门螺杆菌标准株26695的基因进行定点突变,利用重叠延伸PCR法使1398和1399位点的AG突变为GC,获得重组hsp60基因片段并构建原核表达质粒pEASY-E1-rhsp60,经IPTG诱导在大肠杆菌表达突变型rhsp60,将50、100、200μg／mL的rhsp60（rhsp60组）、hsp60（hsp60组）分别与U937细胞共培养,于12、18、24h收集上清液,ELISA法检测上清中细胞因子IL-6水平,设PBS对照组。结果rhsp60组、hsp60组IL-6分泌水平均明显高于对照组（P均〈0．05）,且同时点同浓度下rhsp60组IL-6分泌水平明显高于hsp60组（P均〈0．05）,IL-6的分泌随着蛋白刺激浓度的升高而增高。结论rhsp60与hsp60皆可促进U937细胞分泌IL-6,胃癌来源的rhsp60使IL-6分泌量更多。     

肺结核患者血浆中热休克蛋白70的表达及意义

目的通过检测肺结核患者外周血中热休克蛋白70(HSP70)的表达情况,旨在探讨HSP70与肺结核的关系。方法采用双抗体夹心ELISA法检测32例活动性肺结核组患者外周血中HSP70蛋白的表达情况,并与25例稳定性肺结核组及30例健康对照组进行对照分析。结果活动性肺结核患者血浆中HSP70蛋白表达水平为(7.5±1.3)ng/ml,显著高于稳定性肺结核组(5.5±1.7)ng/ml及健康对照组(5.3±0.8)ng/m(P<0.01);稳定性肺结核组与健康对照组比较差异无显著性(P>0.05)。结论活动性肺结核患者存在HSP70的过度表达,提示HSP70可能通过某些未明机制参与活动性肺结核的发病,并可辅助肺结核活动性的判断。     

Dynamics of heat shock protein 60 in endothelial cells exposed to cigarette smoke extract

Heat shock protein 60 (HSP60), expressed on the surface of endothelial cells (ECs) stressed by e.g. oxidized LDL or mechanical shear, was shown to function as an auto-antigen and thus as a pro-atherosclerotic molecule. The aim of this study was to determine whether cigarette smoke chemicals can lead to the activation of the “HSP60 pathway.” It was also our aim to elucidate the dynamics of HSP60 from gene expression to endothelial surface expression and secretion. Here we show for the first time that the exposure of human umbilical vein endothelial cells (HUVECs) to cigarette smoke extract (CSE) results in an up-regulation of HSP60 mRNA. Live cell imaging analysis of a HSP60-EYFP fusion protein construct transfected into ECs revealed that mitochondrial structures collapse in response to CSE exposure. As a result, HSP60 is released from the mitochondria, transported to the cell surface, and released into the cell culture supernatant. Analysis of HSP60 in the sera of healthy young individuals exposed to secondhand smoke revealed significantly elevated levels of HSP60. Cigarette smoking is one of the most relevant risk factors for atherosclerosis. Herein, we provide evidence that cigarette smoke may initiate atherosclerosis in the sense of the “auto-immune hypothesis of atherosclerosis.”     

The 60-kDa heat shock protein regulates energy rearrangement and protein synthesis to promote proliferation of multiple myeloma cells

To investigate the cellular mechanisms of multiple myeloma (MM), we used liquid chromatography-tandem mass spectrometry for proteomics analysis of CD138⁺ plasma cells from patients with MM and healthy controls. We found that the 60-kDa heat shock protein (HSP60, also known as HSPD1) was significantly upregulated in myeloma cells. HSP60 is an important chaperone protein that regulates the homeostasis of mitochondrial proteins and maintains mitochondrial function. Knockdown (KD) of HSP60 in myeloma cells resulted in inhibition of proliferation and reduced the quality of the mitochondria. Mitochondrial stress tests showed that HSP60 KD inhibited glycolysis and mitochondrial activity. Metabolomics showed a decrease in glycolysis and tricarboxylic acid cycle metabolites, and inhibited the formation of creatine and phosphocreatine by the reaction of S-adenosylmethionine (SAM) with amino acids mediated by demethyladenosine transferase 1, mitochondrial (TFB1M) and reduced energy storage substances. Moreover, HSP60 silencing influenced the synthesis of ribonucleotides and nicotinamide adenine dinucleotide phosphate (NADPH) by the pentose phosphate pathway to inhibit cell proliferation. HSP60 KD inhibited 5' adenosine monophosphate-activated protein kinase (AMPK), which inhibited the key enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), effecting the metabolism of fatty acids by inhibiting malonyl-coenzyme A. Our data suggest that reduced HSP60 expression alters metabolic reprogramming in MM, inhibits tumour progression and reduces mitochondrial-dependent biosynthesis, suggesting that HSP60 is a potential therapeutic target for MM treatment.     

热休克蛋白70对应激性胃溃疡黏膜的保护作用

目的:探讨热休克蛋白70(HSP70)表达是否随应激性胃渍疡病程的进展而变化.方法:收集30例应激性胃溃疡出血患者及10例术前患者的胃液及外周血,采用TRIzol一步法提取胃液脱落细胞及外周血白细胞总RNA,采用半定量RT-PCR检测HSP70的表达;同时,采用免疫组化技术检测外周血白细胞HSP70的表达.结果:RT-PCR检测结果提示应激性溃疡各期的HSP70表达明显高于对照组的表达(血标本:1期0.6289±0.1839,2期1.1322±0.3683,3期0.6855±0.1923,对照组0.3868±0.2071;胃液标本:1期0.7741±0.2442,2期1.2385±0.4558,3期0.8790±0.2722,对照组0.4626±0.2416:均P＜0.05),应激性溃疡2期HSP70表达显著高于1期和3期(P＜0.05),而1期和3期之间无统计学差异(P＞0.05).免疫组化结果为应激性溃疡患者外周血白细胞HSP70的表达水平在不同病期有所不同(1期40%,2期80%,3期30%,对照组10%,χ2=10.8383,P＜0.05),以2期表达水平最高,对照组表达最低.结论:应激性溃疡时患者外周血细胞和胃液脱落细胞均可检测到HSP70的表达,应激状态可诱导HSP70的表达,HSP70对胃黏膜具有保护作用,并可能促进应激性溃疡的愈合.     

热激蛋白70(HSP70)及血吸虫HSP70研究进展

热激蛋白(heat shock proteins,HSP)是细胞或生物体受到外界各种刺激时产生的一种具有重要生物学功能和具有高度保守性的多肽类蛋白质分子家族,其中HSP70是最受关注且研究较为深入的一种.该文阐述了HSP及血吸虫HSP70的研究进展,并对其研究意义作一综述.     

兔动脉粥样硬化易损斑块中高表达热激蛋白70细胞与细胞凋亡的关系

目的:探讨热激蛋白(HSP)的高表达和细胞凋亡在兔动脉粥样硬化斑块中的作用。方法:取兔动脉粥样硬化易损斑块28例,镜下将斑块分为4个区,分别为纤维帽区、脂核区、肩部区和中膜平滑肌区;采用免疫组化技术检测各区不同细胞HSP70的高表达情况、原位末端标记法(TUNEL)检测各区不同细胞凋亡情况。结果:纤维帽区与肩部区HSP70的表达阳性炎性细胞数均显著高于脂核区、中膜平滑区,纤维帽区平滑肌细胞HSP70的阳性表达显著高于其他3个区。脂核区TUNEL阳性细胞数明显高于其他3个区。纤维帽区与肩部中表达HSP70的阳性细胞数大于TUNEL阳性细胞数(P<0.05,P<0.01);脂核区内TUNEL阳性细胞数明显大于表达HSP70的阳性细胞数(P<0.01)。结论:易损斑块内HSP70的高表达主要在纤维帽区及肩部区,而细胞凋亡主要在斑块脂核周围。     

Regulation of 60-kDa heat shock protein expression by systemic stress and 5-hydroxytryptamine in rat colonic mucosa

Bowel dysfunction such as irritable bowel syndrome caused by stress is well described. Previous reports suggest that 5-hydroxytryptamine (5-HT) mediates alteration of bowel motility. In this study, the effects of water-immersion stress and the administration of 5-HT on the expression of a 60-kDa heat shock protein (HSP60) in rat colonic mucosa were investigated. The effect of YM-060, a 5-hydroxytryptamine 3 (5-HT₃) receptor antagonist, on the expression of this protein was also studied. Water-immersion stress and the administration of 5-HT induced synthesis of HSP60 in rat colonic mucosa. The induction of HSP60 and the number of defecations were clearly inhibited by the oral administration of YM-060. Our results suggest that the induction of HSP60 in rat colonic mucosa by water-immersion stress may be associated with gastrointestinal motility mediated by 5-HT, especially via 5-HT₃ receptors.     

艾灸足三里和梁门穴诱导热休克蛋白70抗大鼠胃黏膜氧化损伤作用

目的:观察艾灸足三里和梁门穴对应激性溃疡大鼠胃黏膜热休克蛋白70(HSP70)表达的影响,探讨艾灸足阳明经穴抗胃黏膜氧化损伤的作用机制.方法:将SD大鼠60只完全随机平均分为空白组、模型组、艾灸足三里等穴组和艾灸非穴对照点组,采用水浸-束缚应激法制备应激性溃疡模型.按Guth法计算胃黏膜损伤指数(UI),用激光多普勒血流仪测定胃黏膜血流量(GMBF),用免疫组织化学法和硫代巴比妥酸染色法对处理后大鼠检测其胃黏膜HSP70的表达和丙二醛(MDA)的含量.结果:与模型组和艾灸非穴组比较,艾灸足三里等穴可使应激性溃疡大鼠胃黏膜损伤指数明显下降(14.100±5.425vs26.800±9.807,26.200±7.729,P<0.01),HSP70表达上调(0.133±0.035vs0.077±0.057,0.059±0.038,P<0.01)、血流量增高(279.827±172.862mL/minvs139.489±33.133,141.512±58.450mL/min,P<0.05)、MDA含量减少(2.586±0.252μmol/Lvs3.906±0.768,3.464±1.502μmol/L,P<0.05).结论:艾灸足三里和梁门穴能诱导胃黏膜HSP70高表达并降低MDA含量,以达到其抗氧化损伤作用,并有相对的穴位特异性.     

Expression and significance of heat shock protein 70 and glucose-regulated protein 94 in human esophageal carcinoma

AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageai carcinoma and adjacent normal tissues. METHODS: The expression of HSP70 and grp94 in 78 human esophageai cancer and adjacent normal tissues was studied by immunohistochemistry and pathology photograph analysis. RESULTS: Both esophageai cancer and adjacent normal tissues could express HSP70 and grp94. Of the 78 cases of esophageai carcinoma, 95.0%(72/78) showed positive HSP70, mainly stained in nuclei, while grp94 was mainly stained in cell plasma, and the positive rate was 71.8% (56/78).There was a significant difference in the expression of HSP70 and grp94 between esophageai cancer and adjacent normal tissues (P<0.01). Compared with adjacent normal tissues, there was a significant difference between differential types and HSP70 expression (P<0.01). CONCLUSION: HSP70 and grp94 express differently in cell plasma and nuclei. The expression intensity of HSP70 is related to the differentiation of esophageai carcinoma.