HSP72在视网膜的表达及其对抗细胞凋亡作用的研究

目的：检测锌离子诱导的HSP72在大鼠视网膜的表达及HSP72对视网膜缺血再灌注损伤所致细胞病理性凋亡的抑制作用。方法：生理盐水前房加压灌注制作视网膜缺血再灌注（RIR）损伤模型．以腹腔注射硫酸锌诱导HSP72表达作为实验组,以腹腔注射HSP表达抑制荆——槲皮素作为实验对照组．以不作任何处理的大鼠为正常对照组。于不同时间段对视网膜HSP72免疫组化染色．Tunel染色、计数凋亡细胞,比较各组差异。结果：实验组在注射硫酸锌后10h即见视网膜节细胞层有HSP72阳性表达．16h达高峰,注射后7d仍呈弱阳性表达,HSP72主要在神经节细胞（RGC）胞浆表达;正常对照组及实验对照组均呈阴性表达。Tunel染色显示,RIR后12h即有病理性细胞凋亡现象,24h明显增多,实验组凋亡细胞计数少于对照组且有统计学意义（P〈0．05）。结论：（1）腹腔注射锌离子可诱导大鼠视网膜HSP72表达．注射槲皮素可抑制此作用,HSP72主要在RGC胞浆表达;（2）RIR能导致视网膜病理性细胞凋亡．HSP72具有对抗凋亡的作用。     

HSP9Oα、HSP7O在喉癌中的表达和临床意义

研究HSP90α、HSP70在喉癌中的表达与预后的关系。方法用免疫组化方法检测46例喉 鳞状细胞癌石蜡包埋标本的HSP90a、HSP70。结果喉癌鳞状细胞中HSP90a、HSP70阳性率分别为69.56％ 和67.34％,HSP90α、HSP70的表达与病理分级和预后有关。结论HSP90α、HSP70可作为判断患者预后的 独立指标。     

热休克蛋白70与头颈部肿瘤

热休克蛋白70 （heat shock protein 70,HSP70）是细胞受应激原刺激后诱导所产生的一种应激蛋白,它在进化上高度保守,具有多种生物学功能,包括分子伴侣、细胞保护及抗细胞凋亡的功能,参与多种生理病理过程.近年研究发现HSP70与肿瘤的发生、发展、预后以及机体对肿瘤治疗药物耐药性的发生等都有密切关系,而头颈部肿瘤中也有大量的HSP70表达.本文以HSP70与肿瘤发生、发展的最新进展做一综述,探讨头颈部肿瘤与HSP70的关系.     

鼻咽癌HSP70和HSP90β的表达及临床意义

目的:探讨鼻咽癌组织中HSP70和HSP90β的表达及其与组织分级、预后之间的关系。方法:应用免疫组织化学SP法分别检测50例鼻咽癌组织中HSP70和HSP90β的表达情况,并分析二者表达与鼻咽癌临床与预后的关系。另取10例鼻咽部正常组织作为正常对照。结果:鼻咽癌中HSP70和HSP90β阳性率高于正常组织,而且鼻咽癌分化程度越低,HSP70和HSP90β的表达越强;HSP90β和HSP70阴性表达者5年生存率明显高于阳性者(P<0.05)。结论:HSP90β和HSP70的过度表达与鼻咽癌的发生、发展以及预后有关。     

榄香烯联合热疗对Hep-2细胞周期进程及凋亡率的实验观察

目的观察榄香烯联合热疗对体外培养的Hep-2细胞株（人喉癌细胞株）增殖抑制作用。方法应用四甲基偶氮唑蓝（MTT）比色法检测不同组别对Hep-2细胞增殖抑制率,流式细胞仪检测Hep-2细胞周期进程的变化及凋亡率,透射电子显微镜观察Hep-2细胞亚细胞水平变化。结果单纯榄香烯组、单纯热疗组及榄香烯联合热疗组,对Hep-2细胞增殖抑制率分别为28.8%、25.7%和47.1%,榄香烯联合热疗组能显著提高Hep-2细胞增殖抑制率。细胞周期进程发生明显改变：G1期细胞增多,S期细胞减少,G2/M期细胞相对增多,凋亡率上升。透射电子显微镜下可见线粒体肿胀,核染色质趋边凝集及凋亡小体。结论榄香烯联合热疗能够抑制Hep-2细胞增殖,抑制Hep-2细胞G1期向S期转化进程,诱导Hep-2细胞凋亡。     

热休克蛋白90β(HSP90β)真核表达载体的构建及体外表达

目的克隆人热休克蛋白90β(HSP90β)基因,构建其真核表达载体pcDNA3.1( )/hHSP90β,并检测其在体外的表达情况,为下一步研究鼻咽癌中HSP90β功能奠定基础。方法提取鼻咽癌总RNA,并经RT-PCR获得hHSP90βcDNA。用纯化的hHSP90βcDNA与PGEM Easy T Vector连接,构建中间载体PGEM-hHSP90β。将PGEM-hHSP90β及pcDNA3.1( )质粒经AflII和Xbal双酶切、胶回收纯化酶切片段后,体外连接酶切片段,构建hHSP90β真核表达载体pcDNA3.1( )/hHSP90β。经酶切和测序后,用脂质体包裹转染COS细胞,采用RT-PCR和Western blot检测HSP90β的表达。结果酶切及测序鉴定表明,hHSP90β与pcDNA3.1( )体外重组成功,命名为pcDNA3.1( )/hHSP90β。该表达质粒在体外转染COS细胞后可表达HSP90β分子。结论采用体外重组技术,成功地将人HSP90β插到了真核表达载体pcDNA3.1( )中;pcDNA3.1( )/hHSP90β表达质粒能在体外表达HSP90β分子。     

热休克蛋白72在青光眼视网膜神经节细胞中的保护机制研究进展

青光眼是以不可逆性、进行性视功能损害为特征的视神经退行性病变。研究认为,保护视网膜神经节细胞,使视神经免于继续受损已成为青光眼研究的重点。热休克蛋白72(HSP72)是热休克蛋白家族重要成员之一,是凋亡的抑制者,与神经保护有密切关系。研究HSP72在青光眼视网膜神经节细胞中的抗凋亡和细胞保护特性,对青光眼发病机制的理论研究和临床治疗具有重要的指导意义。就HSP72在青光眼视网膜神经节细胞中的保护作用进行综述。     

中耳细菌感染诱导的急性期HSP—70相关反应的免疫组织化学研究

探讨中耳细菌感染急性期时,哺乳动物中、内耳热休克反应的部位,以及中耳细菌感染诱生的热休克蛋白是否可能引发内耳自身免疫损伤。运用中耳注射肺炎克雷伯杆菌制成豚鼠中耳急性感染动物模型,分别于接种后第1、3、5、7天处死动物取材。应用免疫组化技术,研究了中耳粘膜和耳蜗表达热休克蛋白70（HSP－70）的部位。结果表明：正常状态下,中耳粘膜表层的上皮细胞和内耳膜迷路血管纹、螺旋韧带、Corti氏器均有弱的阳性反应,感染应激后,上述同样部位均有强的阳性显色。不同取材时段显示的阳性位置无差异。说明在中耳急性细菌感染期,中耳粘膜和内耳组织均表达了同源HSP－70蛋白分子,这些同源HSP－70为引发内耳自身免疫损伤提供了物质基础。     

实验性急性化脓性中耳炎中耳粘膜热休克蛋白的表达

目的 :探讨急性化脓性中耳炎时中耳粘膜发生的热休克反应。方法 :向豚鼠中耳注射肺炎克雷伯杆菌 ,制成中耳急性感染动物模型 ,分别于接种后第 1、3、5、7天处死动物取材。应用免疫转印技术动态分析中耳粘膜热休克蛋白 ( HSP)的表达。结果 :正常非应激状态下 ,豚鼠中耳粘膜仅有很弱的 HSP70的表达 ;中耳炎急性期 ,中耳粘膜 HSP70的表达明显升高 ,以第 3天为最强 ,第 3天开始表达 31k D分子 ,出现两条带 ;第 5天最强 ,至第 7天消失。 17k D处第 1～ 5天均有两条带 ,第 3天最高 ,第 7天减弱。结论 :观察发现 ,肺炎克雷伯杆菌感染豚鼠中耳后 ,中耳粘膜出现热休克反应 ,表达 HSP70及与之有相关表位的 HSP31k D和 HSP17k D分子     

人类嗅觉受体神经元的热休克蛋白70表达:年龄相关的趋势

热休克蛋白（heatshockprotein，HSP）是一组进化上高度保留的蛋白质，在机体正常生化过程中起重要作用。其中报道最多的HSP70。在应激状态下，其合成会显著增加，而增加的HSP水平对面临损伤的细胞具有保护作用。曾有学者报道过有关人类各种组织中的HSP免疫反应。该文用免疫细胞化学的方法研究了人类鼻粘膜HSP70表达的细胞分布及其与年龄的相关性。研究人群中共22具尸体，其年龄范围是出生前16周至90岁，包括3例Alzheimer’s病患者。研究表明，HSP70免疫反应性存在于嗅受体神经元、嗅上皮支持细胞的核上区及固有层Bowmann’s腺的腺…     

Study on the expression of HSP70 and HSP90beta in nasopharyngeal carcinoma and the clinical significance]

OBJECTIVE: To study the expression of HSP70 and HSP90beta and its clinical significance in human nasopharyngeal carcinom. METHOD: Immunohistochemical method SP was used to detect the expression of HSP70 and HSP90beta in 50 cases and its clinical significance were studied. RESULT: The positive rate of HSP70 and HSP90beta were 72% and 56% respectively. Analysis of patients, survival demonstrated that the prognosis of NPC with HSP70 (-) and HSP90beta (-) expression were significantly better than the others. CONCLUSION: The overexpression of HSP70 and HSP90beta were probably concerned with the occurrence, development and prognosis of nasopharyngeal carcinoma.     

Upregulation of HSP by geranylgeranylacetone protects the cochlear lateral wall from endotoxin-induced inflammation

We investigated whether an acyclic polyisoprenoid antiulcer drug, geranylgeranylacetone (GGA), induces the expression of HSP70 in the rat cochlea. Immunoblotting revealed upregulation of HSP70 in the cochlea at 12 h after transtympanic (local) or oral (systemic) administration of GGA, and this increased at 24 h after administration. Positive immunohistochemical staining of HSP70 was observed in the hair cells, the spiral ganglion, the stria vascularis, the spiral ligament, and the perivascular portion of modiolar vessels. We therefore subsequently studied the effects of GGA as an HSP-inducer on inner ear trauma due to inflammation. Damage to the lateral wall due to inflammation induced by lipopolysaccharide inoculation was protected against by pretreatment with GGA, as assessed physiologically by measurement of cochlear blood flow and morphologically by electron microscopy. The results of the present study suggest that GGA can protect the cochlea against other injuries including those induced by noise, ototoxic drugs, and ischemia by upregulating HSP70.     

Expression of HSP70 and Its Relation with Other Cytokines in Human Middle Ear Effusion

Objectives

While other cytokines are known to be associated with otitis media with effusion (OME), the involvement of heat shock protein 70 (HSP70) in middle ear effusion (MEE) is unknown. This study was undertaken to investigate the possibility of there being a HSP70 expression in human MEE and to determine its potential role as a cytokine in OME.

Methods

The levels of HSP70, tumor necrosis factor-alpha and interleukin-1beta were measured by enzyme-linked immunosorbent assay in the effusion of different groups of OME patient following collection of the MEE using our new collection system. The clinical characteristics of the OME patients and the MEE status were analyzed.

Results

HSP70 was expressed in all the types of MEE. The mucous and seromucous effusions showed higher HSP70 levels than that of the serous effusion. The HSP70 level was correlated with the levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in the effusions. The positive correlations between HSP70, TNF-α and IL-1β were statistically significant (P<0.05).

Conclusion

The highly elevated level of HSP70 in the seromucous and mucous effusions implicates this protein in the chronicity of OME.     

Expression of heat shock protein (HSP70) in oral lichen planus and non-dysplastic oral leucoplakia

The purpose of this study was to investigate the expression of heat shock protein (HSP70) in oral non-dysplastic leucoplakia and in relation to the clinical and pathological features of oral lichen planus. The expression of HSP70 was assessed in the epithelial compartment of normal mucosa (n = 5), oral lichen planus (n = 28) and benign leucoplakia (n = 11) using an immunohistochemical method. The immunostaining intensity distribution (IID) index was used to quantify the positivity of the staining. There was no association between HSP70 overexpression and clinical presentation of oral lichen planus. Oral lichen planus patients showed no statistically significant differences in the depth of the inflammatory infiltrate when expression of HSP70 was considered (X(i)- X(j) = 42.30; 95% confidence interval (95% CI) = -120.87-205.48). No statistically significant differences were identified in terms of HSP70 expression between oral lichen planus and normal buccal mucosal specimens (X(i)- X(j) = 4.07; 95% CI = -0.53-8.67). The IID index score for HSP70 expression in leucoplakia specimens was significantly higher than the one of the oral lichen planus group (X(i) - X(j)= 5.11; 95% CI = 1.73-8.48). It is concluded that there are no statistically significant differences in HSP70 expression between oral lichen planus and normal buccal mucosal specimens, suggesting that HSP70 does not play an obvious part in the pathogenesis of oral lichen planus. The expression of HSP70 was significantly higher in oral leucoplakia than in oral lichen planus, possibly because of differences in cellular activity or cell proliferation.     

喉癌组织中热休克蛋白和人乳头状瘤病毒的相关性

目的 探讨喉鳞状细胞癌（简称喉癌）组织中热休克蛋白70（heat shock protein 70,HSP70）和人乳头状瘤病毒1 6E7（human papiloma-virus 1 6E7,HPV16E7）蛋白的表达及其在喉癌发生和发展过程中的相互关系.方法 用免疫组织化学方法检测78例喉癌、24例声带息肉和10例癌周喉组织标本中HSP70和HPV16E7蛋白的表达.结果 在喉癌组织、声带息肉和癌周喉组织中HSP70的阳性表达率分别为69.2%,8.3%和0%,HPV16E7蛋白的阳性表达率分别为43.6%,4.2%和0%,HSP70和HPV16E7蛋白的阳性表达在喉癌组织与声带息肉、癌周喉组织之间均有显著性差异（P＜0.05）;HPV16E7阳性的喉癌组织中HSP70的阳性表达率为85.3%,HPV16E7阴性的喉癌组织中HSP70的阳性表达率为56.8%,二者之间有显著性差异（P＜0.05）.结论 HPV16E7可以诱导喉癌组织中HSP70的高表达,二者在喉癌的发生和发展过程中发挥重要作用;检测HSP70和HPV16E7蛋白的表达可用于喉癌恶性程度的评定.     

Expression of heat shock protein (HSP70) in oral lichen planus and non‐dysplastic oral leucoplakia

Expression of heat shock protein (HSP70) in oral lichen planus and non‐dysplastic oral leucoplakia The purpose of this study was to investigate the expression of heat shock protein (HSP70) in oral non‐dysplastic leucoplakia and in relation to the clinical and pathological features of oral lichen planus. The expression of HSP70 was assessed in the epithelial compartment of normal mucosa (n = 5), oral lichen planus (n = 28) and benign leucoplakia (n = 11) using an immunohistochemical method. The immunostaining intensity distribution (IID) index was used to quantify the positivity of the staining. There was no association between HSP70 overexpression and clinical presentation of oral lichen planus. Oral lichen planus patients showed no statistically significant differences in the depth of the inflammatory infiltrate when expression of HSP70 was considered ( X?_{i ? X?j = 42.30; 95% confidence interval (95% CI) = ?120.87–205.48). No statistically significant differences were identified in terms of HSP70 expression between oral lichen planus and normal buccal mucosal specimens ( X?i ? X?j = 4.07; 95% CI = ?0.53–8.67). The IID index score for HSP70 expression in leucoplakia specimens was significantly higher than the one of the oral lichen planus group ( X?i ? X?j} = 5.11; 95% CI = 1.73–8.48). It is concluded that there are no statistically significant differences in HSP70 expression between oral lichen planus and normal buccal mucosal specimens, suggesting that HSP70 does not play an obvious part in the pathogenesis of oral lichen planus. The expression of HSP70 was significantly higher in oral leucoplakia than in oral lichen planus, possibly because of differences in cellular activity or cell proliferation.     

Immunohistochemical assesment of heat shock protein 70 in adenoid tissue

OBJECTIVES: To evaluate the expression of heat shock protein 70 (HSP70) and its relation to histopathologic parameters in adenoid hypertrophy and hyperplasia. In addition, HSP70 expression in adenoid tissue was compared with in adult and childhood. METHODS: Paraffin-embedded adenoid tissue sections were obtained from 19 childhood and 15 adult patients. Expression of HSP70 was evaluated by immunohistochemical staining using anti-HSP70 monoclonal antibody and correlated with histopathologic parameters. RESULTS: Positive HSP70 expression was observed mainly in the mucosal epithelium, lymphocytes in germinal centers, interfollicular lymphocytes, subepithelial plasma cells and vascular endothelium. HSP70 immunoreactivity in the mucosal epithelium with severe intraepithelial lymphocytic infiltration in childhood patients was higher than in adult patients. Although, the immunoreactivity of HSP70 was detected strongly in whole layer of metaplastic squamous epithelium, it was stained only in basal layers in respiratuary epithelium, Lymphocytes stained with HSP70 in germinal center and interfollicular areas of adenoid tissues was higher in childhood patients than in adults. CONCLUSIONS: These data suggest that HSP70 expression may have an important role in pathogenesis of adenoid hyperplasia, especially, in childhood.     

Expression of heat shock protein, HSP72, in the guinea pig and rat cochlea after hyperthermia: immunochemical and in situ hybridization analysis.

The induction of the heat shock protein, HSP72, was studied in the cochlea of guinea pigs and rats subjected to a hyperthermic stress. Analyses were done by immunoblotting and immunocytochemistry at 6 and 12 h after heat shock, using a commercially available monoclonal antibody (Amersham), and by in situ hybridization 1 h after heat shock using an oligonucleotide probe. In guinea pig immunoblots of the cochlea, HSP72 was present in both unstressed and heat stressed animals and immunocytochemistry did not reveal any difference of staining between them. As opposed to guinea pig, HSP72 was not found in unstressed rat cochlea. Heat shock induced HSP72 expression in most inner ear tissues of the rat examined by immunoblotting. Immunocytochemistry and in situ hybridization localized HSP72 synthesis in ganglion neurons, Schwann cells, spiral limbus, spiral ligament and stria vascularis. The strongest immunoreactivity and highest density of silver grains were seen in the stria vascularis. All blood vessels were strongly immunoreactive and were outlined with silver grains. These results show that HSP72 synthesis can be induced by hyperthermia in rat cochlea and suggest that this protein could be a useful marker for assessment of the effects of specific stresses in this organ.