热休克蛋白27、热休克蛋白65和肽素在青年缺血性脑卒中患者血清中的表达及其临床意义

目的 探讨热休克蛋白(HSP) 27、HSP65和肽素在青年缺血性脑卒中患者血清中的表达及其临床意义.方法 选取135例体检合格的健康人群为对照组,124例青年缺血性卒中患者为实验组,采用酶联免疫吸附试验(ELISA)检测HSP27、HSP65和肽素表达水平的变化.结果 青年缺血性脑卒中的发病与患者的年龄、身高、体重和身体质量指数(BMI)没有显著的关系(F=26.38,P＞0.05).青年缺血性脑卒中患者和对照组相比,患者血清中HSP27、HSP65和肽素的表达水平显著升高(F=55.71,P＜0.05);女性脑卒中患者和男性脑卒中患者相比,患者血清中HSP27和肽素的表达水平较高,而HSP65表达水平较低(F=62.65,P＜0.05).结论 HSP27、HSP65和肽素可能是青年缺血性脑卒中的危险因素.     

应激相关的热休克信号转导通路的研究进展

随着社会的发展和医疗技术的不断进步与完善,人们的健康水平有了大幅度的提高。但急性梗塞(如脑梗、心梗)、感染性疾病(如败血症)等仍有较高的发病率和死亡率。特别是随着我国社会和经济的发展,老年人群的结构比例明显增高,心脑血管疾病和感染引起的疾病将进一步增加。而且随着     

Heat shock protein 27 expression in patients with chronic liver damage

The aim of this study was to evaluate a possible relationship between lymphomonocyte expression of heat shock proteins (HSP) 60/27 and plasma levels of pro-inflammatory cytokines (tumor necrosis factor-alpha and interleukin-6) and markers of antioxidant/oxidative status [glutathione (GSH), alpha glutathione-S-transferase activity (alpha GST), malonyldialdeyde (MDA), 4-hydroxinonenal (4-HNE), and S-nitrosothiols (S-NO)] in patients with chronic liver diseases. Entered into the study were 47 subjects: 10 healthy controls, 16 patients with HCV-related chronic hepatitis (CH), and 16 patients with HCV-related and 5 with alcohol-related liver cirrhosis (10 Child A and 11 Child B+C). HSP60 was clearly expressed only in 5% of patients and lowly in the control group. HSP27 was clearly expressed in 46.7% of CH and 71.4% of cirrhotic patients but was lowly present in healthy subjects. A significant difference was found between patients with a low expression of HSP27 (negative patients) and those with a high HSP27 expression (positive patients) of plasma levels both of antioxidants (GSH, p < 0.05), and of markers of enhanced production of free radicals and cytokines (alpha GST, TNF-alpha and IL-6, p < 0.05; MDA, 4-HNE and S-NO, p < 0.01) as well as for alcohol use and degree of liver impairment. The present data are the first showing that, particularly in conditions of enhanced oxidative stress, lymphomonocytes from liver disease patients present an increased expression of HSP27.     

热休克蛋白在人乳腺癌中的作用

目的探讨不同类型和亚型热休克蛋白（Ｈｓｐｓ）在人乳腺癌增殖、分化中的作用及意义。方法应用免疫组织化学ＳＰ法、电镜、原位杂交和ＲＴ－ＰＣＲ法对１１５例浸润性导管癌Ｈｓｐ７０、Ｈｓｐ９０、泛素（ｕｂｉｑｕｉｔｉｎ）进行观察。结果Ｈｓｐ９０ｍＲＮＡ在癌组织较非癌组织中的表达显著增强。此外，癌组织中的Ｈｓｐ９０αｍＲＮＡ表达水平与增殖细胞核抗原（ＰＣＮＡ）指数密切相关。提示Ｈｓｐ９０α同功异构体表达增加在细胞增殖过程中起一定作用。另外，在分化差的癌组织中也有Ｈｓｐ９０βｍＲＮＡ表达。Ｈｓｐ７０和泛素在细胞中的分布部位一致。在Ｈｓｐ７０核内染色阳性的标本中，ＰＣＮＡ指数显著增加。Ｈｓｐ７０家族中的Ｈｓｐ７３ｍＲＮＡ在ＰＣＮＡ高表达的癌组织比非癌组织有较高水平的表达。结论本研究显示：Ｈｓｐ９０α在癌细胞增殖中起重要作用，而Ｈｓｐ９０β与细胞分化和结构构建有关。此外，Ｈｓｐ７０家族中，尤其是与泛素有关的Ｈｓｐ７３可能是癌细胞增殖的一个指标。     

Induction of heat shock protein 72 synthesis by endogenous tumor necrosis factor via enhancement of the heat shock element-binding activity of heat shock factor 1

Endogenous tumor necrosis factor (enTNF) acts as a resistance factor against cytotoxicity caused by heat by inducing manganous superoxide dismutase (MnSOD), thereby scavenging reactive oxygen free radicals. On the other hand, it is also well known that heat shock proteins (HSP) which are induced by heat stress behave as cytoprotective factor against this stress. However, the relationship of these two resistance factors is not elucidated yet. In the present study, we therefore proposed the possibility that enTNF enhances HSP72 expression. Heat-sensitive L-M (mouse tumorigenic fibroblast) cells, which normally do not express enTNF, were transfected with a nonsecretory-type human TNF-α expression vector to produce enTNF. Stable transfectants showed resistance to heat treatment and an increase of HSP72 expression. Conversely, when HeLa (human uterine cervical cancer) cells, which normally produce an appreciable amount of enTNF, were transfected with an antisense TNF-α mRNA expression vector to inhibit enTNF synthesis, their heat sensitivity was enhanced and HSP72 expression was reduced by half. Although enTNF caused no difference in the level of heat shock factor (HSF) 1 in these cells, enTNF expression correlated well with the binding activity of HSF-1 to a ³²P-labeled synthetic oligonucleotide containing the human heat shock element (HSE). These results indicate that enTNF participates not only in intrinsic resistance against heat via induction of MnSOD but also via enhancement of the HSE-binding activity of HSF 1 followed by augmentation of HSP72 expression.     

热休克蛋白对树突状细胞成熟的影响研究

目的:探讨热休克蛋白(HSP)对树突状细胞(DCs)成熟的影响,同时对其形态学动态变化进行研究。方法:从肝癌组织中提取HSP(gp96 )抗原肽复合物;肝癌细胞进行热休克处理诱导其表面表达HSP,再用DiI荧光标记;将两种HSP与DCs混合培养,动态观察DCs捕获抗原和成熟过程中的形态学变化。流式细胞仪检测不同情况下热休克蛋白对DCs成熟表型的影响。结果:使用DiI标记的方法良好地显示了DCs摄取抗原的形态学变化;DCs通过直接接触、包绕、形成囊泡和伸出伪足且末端形成囊泡4种方式摄取抗原;热休克蛋白可促进DCs成熟表型的表达。结论:DiI是适合DCs形态学研究的良好的荧光标记物;DCs捕获抗原有4种不同方式;不同热休克蛋白均可促进DCs成熟,但提取的热休克蛋白作用更显著。     

Immunohistochemical localisation of the 25 kDa heat shock protein in unstressed rats: Possible functional implications

The distribution of the 25 kDa heat shock protein (hsp 25) in a number of tissue types from unstressed rats was investigated. Immunohistochemical analysis showed that hsp 25 was not found in the thymus, brain (cerebral cortex and cerebellum), testis, adrenal, liver, spleen, or kidney. A number of cells in the anterior pituitary showed strong staining. These cells were tentatively identified as being either gonadotropes or thyrotropes. Strong staining was also observed in the blood vessels within these tissues. Hsp 25 was found to be localised predominantly to intestinal smooth muscle of the duodenum and colon and to vascular smooth muscle. Smooth muscle from other sites, such as the trachea, was also intensely stained. Lower and more variable amounts of staining were observed in cardiac and skeletal muscle. These observations suggest that hsp 25 is associated with cytoskeletal elements in muscle, and that the high staining intensity in smooth muscle might be due to the lack of internal architecture present in this muscle type. © 1993 Wiley-Liss, Inc.     

阿魏酸钠对血管平滑肌细胞热休克蛋白27磷酸化的影响

目的： 研究阿魏酸钠对血管紧张素II（angiotensinⅡ,AngII）与血小板源性生长因子-BB（platelet derived growth factor-BB,PDGF-BB）诱导的血管平滑肌细胞（vascular smooth muscle cells,VSMCs）中热休克蛋白27（heat shock protein 27,HSP27）磷酸化的影响。方法：采用体外培养大鼠胸主动脉平滑肌细胞；用特异性的单克隆phospho-HSP27抗体的蛋白印迹法（Western blotting）检测HSP27的活性。结果：阿魏酸钠以剂量依赖性方式抑制AngII和PDGF-BB诱导的HSP27磷酸化，在10-4 mol/L〖JP2〗时抑制作用最明显，最大抑制率分别为39.0%（P<0.05）和56.8%（P<0.01）。结论：阿魏酸钠具有明显抑制由AngII和PDGF-BB诱导的VSMCs HSP27磷酸化的作用。     

Exogenous heat shock protein 27 uniquely blocks differentiation of monocytes to dendritic cells

Circulating heat shock protein (HSP)-27 is associated with tumor progression and increased post-injury infection. Extracellular HSP-27 might alter monocyte (MO)-derived DC and/or MPhi function to mediate immunosuppression. HSP-27 treatment inhibited expression of CD1a and CD1b/c, antigen uptake, and allogeneic T cell induction (MLR) by IL-4 + GM-CSF-differentiated human DC while increasing some MPhi characteristics ( upward arrowCD14, upward arrowCD16, upward arrowCD163). MO cytokine receptor profiles elicited by 24-h exogenous HSP-27 treatment remained supportive of immature DC (iDC) emergence ( upward arrowIL-4R, downward arrowIL-6R, downward arrowM-CSFR). IL-10, IL-6, and M-CSF (which promote MPhi differentiation) were significantly increased in IL-4 + GM-CSF + HSP-27 MO-->iDC differentiation cultures. However, HSP-27 treatment during MO differentiation to DC increased programmed cell death ligand 1 coinhibitor and depressed CD86 costimulator expression in parallel to decreased iDC MLR activity. This suggested that increased MPhi differentiation was not solely responsible for HSP-27 reduction of differentiating DC activity. HSP-27 treatment actually depressed the phagocytic capacity of MO differentiated to MPhi by IL-10 or M-CSF culture. CD163 (hemoglobin receptor) expression was depressed on M-CSF + HSP-27 MO-derived MPhi. HSP-27-mediated inhibition of MO-->iDC differentiation was reversed by p38alpha & beta inhibitor (SB202190) addition or TLR4 receptor modulation. HSP-27 impaired appropriate MO-->iDC and MO-->MPhi differentiation modulating expression of receptors necessary for their proper functions. This suggests that endogenous HSP-27 has immunoregulatory activities which could contribute to immunopathology.     

Serum heat shock protein 70 level as a biomarker of exceptional longevity

Heat shock proteins are highly conserved proteins that, when produced intracellularly, protect stress exposed cells. In contrast, extracellular heat shock protein 70 (Hsp70) has been shown to have both protective and deleterious effects. In this study, we assessed heat shock protein 70 for its potential role in human longevity. Because of the importance of HSP to disease processes, cellular protection, and inflammation, we hypothesized that: (1) Hsp70 levels in centenarians and centenarian offspring are different from controls and (2) alleles in genes associated with Hsp70 explain these differences. In this cross-sectional study, we assessed serum Hsp70 levels from participants enrolled in either the New England Centenarian Study (NECS) or the Longevity Genes Project (LGP): 87 centenarians (from LGP), 93 centenarian offspring (from NECS), and 126 controls (43 from NECS, 83 from LGP). We also examined genotypic and allelic frequencies of polymorphisms in HSP70-A1A and HSP70-A1B in 347 centenarians (266 from the NECS, 81 from the LGP), 260 NECS centenarian offspring, and 238 controls (NECS: 53 spousal controls and 106 septuagenarian offspring controls; LGP: 79 spousal controls). The adjusted mean serum Hsp70 levels (ng/mL) for the NECS centenarian offspring, LGP centenarians, LGP spousal controls, and NECS controls were 1.05, 1.13, 3.07, 6.93, respectively, suggesting that a low serum Hsp70 level is associated with longevity; however, no genetic associations were found with two SNPs within two hsp70 genes.     

血清热休克蛋白70与妊娠的相关性研究

目的探讨妊娠期间母体与血清热休克蛋白之间的关系。方法用双抗体夹心ELISA法检测114例孕妇血清热休克蛋白含量,按孕3、5、7、8个月和足月共分五组,与非孕对照组比较。结果孕3-8个月组血清HSP70蛋白的含量与非孕对照组比较无差异（P〉0.05）,足月孕妇组血清HSP70含量明显升高,与非孕对照组比较有非常显著性差异（P〈0.001）;重度妊高征患者血清热休克蛋白70含量明显高于同孕龄正常孕妇组（P〈0.001）。结论了解不同孕月健康孕妇血清HSP70含量,可为妊娠期间某些疾病引起的血清HSP70升高,提供参考依据。     

致白内障基因Hsf4b K65R位点突变对下游热休克蛋白表达的影响

目的:探讨致白内障基因热休克因子4b(Hsf4b)K65R位点突变对其调控下游热休克蛋白(HSP)表达的影响。方法:采用KOD-Plus-Mutagenesis-Kit试剂盒构建pWZL-blast-HA-Hsf4b/K65R赖氨酸突变质粒;通过慢病毒感染小鼠晶状体上皮细胞mLEC构建稳定表达Hsf4b/K65R突变质粒的细胞株;Western blotting检测Hsf4b在mLEC K65R点突变细胞株和野生株中的表达;Western blotting及real-time PCR检测K65R点突变后下游蛋白Hsp70、Hsp90、Hsp27和CryAB表达的变化。结果:阳性克隆PCR及基因测序证明慢病毒载体pWZL-blast-HAHsf4b/K65R构建成功。K65R点突变后不影响Hsf4b在小鼠晶状体上皮细胞mLEC中的表达,但能影响下游蛋白CryAB、Hsp27、Hsp70i和Hsp90a的表达。结论:pWZL-blast-HA-Hsf4b/K65R载体可用于慢病毒感染稳定细胞株构建。Hsf4b K65R位点突变能显著影响其对热休克蛋白的调控功能。     

沙眼衣原体感染后热休克蛋白与稽留流产相关性研究

目的探讨沙眼衣原体(CT)感染后热休克蛋白-60(HSP-60)与稽留流产的关系。方法采用荧光定量聚链酶反应(PCR)方法,对72例稽留流产患者和正常人工流产患者进行沙眼衣原体检测,将正常人工流产CT-DNA阴性者随机选择72例作为对照组。再采用免疫组化方法检测蜕膜组织内HSP-60表达,采用HPLAS-1000高清晰度彩色病理图文报告管理系统检测单位面积中各项免疫组化阳性颗粒平均光密度值。结果HSP-60阳性细胞光密度值,A组:116.36±22.63,B组:139.67±32.26,对照组:221.12±29.87;A组HSP-60表达强于对照组,对比有明显差异(P<0.001)A组与B组对比无明显差异(P>0.005)。结论沙眼衣原体感染后,HSP-60蜕膜组织内表达强度与稽留流产呈正相关性;可认为是稽留流产发生的重要原因之一。     

Induction of heat shock protein 27 by hydroxyurea and its relationship to experimental metastasis

Treatment of tumor cells with hydroxyurea (HU) has been shown to increase the experimental metastatic potential of these cells. We have previously described the induction of stress proteins (antioxidants) by in B16 murine melanoma cells and their relationship to the metastatic process. We have now investigated the induction by HU of another set of stress proteins, the heat shock proteins, and their role in experi-mental metastasis. HU markedly increased the cellular content of heat shock protein (hsp) 27 but not hsp 90, 72/73, or 60 as measured by immunoblotting. The induction of hsp27 protein was preceded specific increase in hsp27 mRNA. Furthermore, HU-treated cells were more thermotolerant. To investigate the functional role of hsp27, human hsp27 cDNA was constitutively overexpressed in B16 cells at seen in HU-treated cells. In separate experiments, we induced a global increase in hsps by heat shock. Neither the hsp27 transfectants nor the heat-shocked cells demonstrated an increase in their experimental metastatic capacity. We conclude that hsp27 protein is increased by HU by the specific induction of hsp27 mRNA in B16 melanoma cells but increased hsp27 protein is not responsible for the increase in experimental metastasis. Since high levels of hsp27 are associated with metastatic disease in breast and ovarian cancers, but not in our experimental system, the functional role of hsp27 in metastasis requires further study. © Rapid Science 1998     

睡眠剥夺对HSP70表达及脑超微结构的影响

目的：观察睡眠剥夺（SD）后大鼠脑组织HSP70表达的变化及对超微结构的影响。方法：44只雄性SD大鼠随机分为11组，每组4只，免疫组织化学方法检测HSP70的表达，电镜观察海马超微结构的变化。结果：睡眠剥夺后12小时即可在大脑皮质及海马观察到HSP70阳性细胞，2—3天数量达到高峰，7天时明显下降。白天睡眠剥夺12小时（SDd12h）组HSP70阳性细胞数较夜晚睡眠剥夺12小时（SDn12h）组多（P〈0．05）。RS组大脑皮质HSP70阳性细胞数较白天睡眠剥夺1天（SDd1d）组减少（P〈0．05）。白天睡眠剥夺3天（SDd3d）海马出现超微结构改变，白天睡眠剥夺7天（SDd7d）后改变更加明显。结论：睡眠剥夺可影响大鼠脑组织HSP70表达及超微结构。     

Extracellular heat shock protein 60 (Hsp60) levels in children with septic shock

Objective and design: Recent data suggest that extracellular Hsp60 modulates the host innate immune response. We analyzed plasma Hsp60 levels in children admitted to a level III tertiary care PICU with septic shock. Materials and subjects: Blood samples were obtained from children meeting criteria for septic shock (n = 63), critically ill children without septic shock (n = 10), and healthy controls (n = 24). Treatment: Not applicable. Methods: Hsp60 levels were measured in the plasma using a commercially available ELISA. Differences between groups were analyzed with a Kruskal-Wallis one way ANOVA due to the non-parametric nature of the data. A p value ≤ 0.05 was considered significant. Results: Extracellular Hsp60 levels were significantly higher in children with septic shock (median, 16.7 ng/mL) compared to both critically ill children without septic shock (median, 0 ng/mL) and healthy controls (median, 0 ng/mL, p <0.001). Conclusions: Extracellular Hsp60 levels are significantly elevated in children with septic shock compared with both healthy controls and critically ill children without sepsis. Extracellular Hsp60 may play a role in the pathogenesis of sepsis in children. Received 3 July 2006; returned for revision 18 October 2006; accepted by K. Visvanathan 6 December 2006     

Inhibition of NaCl-induced heat shock protein 72 expression renders MDCK cells susceptible to high urea concentrations

 Exposure of Madin-Darby canine kidney (MDCK) cells to elevated extracellular NaCl concentrations is associated with increased heat shock protein 72 (HSP72) expression and improved survival of these pretreated cells upon exposure to an additional 600 mM urea in the medium. To establish a causal relationship between HSP72 expression and cell protection against high urea concentrations, two approaches to inhibit NaCl-induced HSP72 synthesis prior to exposure to 600 mM urea were employed. First, the highly specific p38 kinase inhibitor SB203580 was added (100 μM) to the hypertonic medium (600 mosm/kg H₂O by NaCl addition, 2 days of exposure), which significantly reduced HSP72 mRNA abundance and HSP72 content. Survival of these cells after a 24-h urea treatment (600 mM) was markedly curtailed compared with appropriate controls. Second, a pcDNA3-based construct, containing 322 bases of the HSP72 open reading frame in antisense orientation and the geneticine resistance gene, was transfected into MDCK cells. Clones with strong inhibition of HSP72 synthesis and others which express the protein at normal levels (comparable to nontransfected MDCK cells) after heat shock treatment or hypertonic stress were established. When these transformants were subjected to hypertonic stress for 2 days prior to exposure to an additional 600 mM urea for 24 h, cell survival was significantly reduced in those clones in which HSP72 expression was strongly inhibited. These results provide further evidence for the protective function of HSP72 against high urea concentrations in renal epithelial cells. Received: 14 September 1998 / Received after revision and accepted: 16 November 1998