脂多糖介导的人牙周膜成纤维细胞损伤过程中TRAF-6蛋白表达的研究

目的观察在不同浓度脂多糖(LPS)干预下,肿瘤坏死因子受体相关分子(TRAF)6在人牙周膜成纤维细胞(HPLFs)损伤过程中表达的情况。方法取原代培养至5～7代的HPLFs,以LPS梯度浓度干预,采用免疫组织化学方法检测TRAF-6在HPLFs中的表达。结果TRAF-6在正常的HPLFs中呈阴性表达,随LPS干预浓度由0.1μg/ml增加到100μg/ml,TRAF-6表达有先逐渐增高然后又降低的趋势。结论TRAF-6在LPS引发的HPLFs炎症损伤中发挥重要的作用,为细胞因子调控网络增添新资料。     

蛋白激酶Cδ亚型抑制剂对脂多糖诱导肺微血管内皮细胞损伤的影响

目的研究蛋白激酶C(PKC)的亚型PKCδ在大鼠肺微血管内皮细胞(RPMVEC)中的表达及其抑制剂Rottlerin对脂多糖(LPS)导致RPMVEC单层通透性增高的影响。方法取体外培养的RPMVEC进行PKCδ的W estern b lot和免疫组化检测,用针头式滤器检测LPS及LPS+Rottlerin干预后RPMVEC单层滤过系数(K f)的变化。结果PKCδ在RPMVEC中有广泛的表达,其表达部位主要位于胞质。Rot-tlerin能减低LPS导致的RPMVEC单层通透性增高。结论LPS导致RPMVEC损伤的机制与PKC的激活有关,PKCδ亚型抑制剂可减轻LPS诱导的RPMVEC单层通透性增高。     

热休克蛋白70在大鼠急性肺损伤中的表达

目的:探讨急性肺损伤(ALI)大鼠肺组织热休克蛋白70(HSP70)的表达。方法:雌性Wistar大鼠36只,随机分为两组:急性肺损伤组(A组)、对照组(B组),大鼠尾静脉注射脂多糖(LPS)复制ALI模型。所有的动物于注射LPS后2,4,6 h处死,测定肺组织HSP70的表达、超氧化物歧化酶(SOD)活性及丙二醛(MDA)的含量。结果:A组同B组相比,各时间点肺组织HSP70表达均增多,以2,6 h升高明显(P<0.01),MDA含量增高(P<0.05),SOD活性明显降低(P<0.05)。结论:HSP70在ALI后的表达增多可能与其参与LPS所致肺损伤后的组织保护有关。     

HSP70和P27在食管鳞癌中的表达及意义

目的探讨热休克蛋白70(HSP70)和P27在食管鳞状细胞癌中表达之间的相关性。方法采用SP免疫组化染色法检测65例食管鳞癌组织、30例正常食管组织及30例瘤旁组织中的HSP70和P27。结果正常食管上皮、癌旁组织、食管鳞癌中HSP70的阳性表达率分别是16.67%(5/30)、46.67%(14/30)、86.15%(56/65),正常食管上皮、癌旁组织及食管鳞癌中P27的表达率分别是96.67%(29/30)、53.33%(16/30)、41.53%(27/65)。在食管鳞癌中,HSP70和P27的表达呈负相关(rs=-0.541,P=0.000)。结论在食管癌中,P27可能随着HSP70表达的上调而下降,其表达率与多个临床病理指标相关。     

MK-801对大鼠脊髓损伤后Caspase-3、HSP27表达的影响

目的探讨大鼠脊髓损伤后应用兴奋性氨基酸受体非竞争性拮抗剂地卓西平马来酸盐(MK-801)对热休克蛋白27(HSP27)、半胱氨酸蛋白酶3(Caspase-3)表达及细胞凋亡的影响。方法 20只SD大鼠平均分为2组,采用Al-len’s法造成SD大鼠脊髓损伤模型,分别给予MK-801及生理盐水治疗,24h后取材,用免疫组化染色检测Caspase-3阳性细胞及HSP27阳性细胞,用流式细胞学检测细胞凋亡。结果两组均发现Caspase-3、HSP27表达及凋亡细胞,损伤组Caspase-3表达及神经细胞凋亡指数均高于MK-801组(P<0.01),而HSP27表达MK-801组高于损伤组(P<0.01)。结论脊髓损伤后,MK-801能上调HSP27的表达,抑制Caspase-3表达及神经细胞凋亡。     

葛根素对急性脑缺血模型大鼠脑细胞损伤后HSP70及Fas蛋白表达的干预作用研究

目的:探讨葛根素对急性脑缺血模型大鼠脑细胞损伤后热休克蛋白70(HSP70)及Fas蛋白表达的干预作用机制。方法:将12只急性脑缺血模型大鼠随机分为单纯缺血组(生理盐水)和葛根素干预组,各6只,每只大鼠再分别按缺血侧(实验组)和非缺血侧(对照组)进行自身对照,用HE染色及免疫组织化学SP法测定HSP70表达情况;另将脑缺血模型大鼠随机分为对照组(生理盐水、缺血侧)、葛根素干预组(缺血侧)、正常组(非缺血侧),各6只,用HE染色及免疫组织化学SP法测定Fas表达情况。结果:单纯缺血对照组及葛根素干预对照组HSP70呈阴性或弱阳性表达,而在单纯缺血实验组及葛根素干预实验组中HSP70均表达增强(P<0.01),葛根素干预实验组的表达则明显强于单纯缺血实验组(P<0.01);Fas蛋白在各组中均有不同程度的表达,阳性细胞绝大部分为神经元,以锥体细胞表达强度最强,各组间阳性细胞数均无显著差异(P>0.05);HE切片中,对照组和葛根素组死亡细胞数均明显多于正常组(P<0.01),且对照组明显多于葛根素组(P<0.01)。结论:葛根素对急性脑缺血损伤具有保护作用,主要通过上调HSP70的表达来实现,而与Fas蛋白的表达及细胞凋亡等相关不强。     

热休克蛋白27在食管上皮癌变过程中的表达及其意义

目的 了解热休克蛋白27(HSP27)在食管鳞癌、癌旁组织和切缘正常食管黏膜中的表达,分析其表达差异,并探讨其与食管癌发生发展的关系.方法 应用免疫组化Envision二步法,观察HSP27在食管鳞癌、癌旁组织和切缘正常食管黏膜中的表达,并比较其间是否存在差异.结果 食管鳞癌、癌旁组织和切缘正常食管黏膜中HSP27的阳性表达率分别为73.3％、53.5％和25.3％;HSP27在食管鳞癌中的阳性表达率高于在癌旁组织(P＜0.05),在癌旁组织中的阳性表达率高于在切缘正常食管黏膜(P ＜0.001).随着食管鳞癌高、中、低分化程度的不同,HSP27的阳性表达率逐渐降低,但差异无统计学意义(P＞0.05).结论 HSP27可能与食管鳞癌的发生发展有关.     

HSP27在甲基强的松龙治疗脊髓损伤中的作用

热休克蛋白27(HSP27)是低分子量热休克蛋白家族(HSPs)中的代表蛋白之一,在甲基强的松龙(MP)治疗脊髓损伤(SCI)中有积极的作用。随着相关研究的深入,了解SCI中HSP27对神经细胞的保护机制,将为临床治疗SCI提供积极的治疗依据。本文对HSP27在MP治疗脊髓损伤中的作用作一综述。     

脂多糖干预后成牙本质细胞系MDPC-23中肿瘤坏死因子受体相关因子6基因表达的研究

目的观察不同浓度脂多糖(LPS)干预后,成牙本质细胞中肿瘤坏死因子受体相关因子6(TRAF6)基因表达的情况。方法体外培养小鼠永生化成牙本质细胞系MDPC-23,采用对照组不干预(即0μg/ml组)、10μg/ml和20μg/mlLPS干预MDPC-23后,通过反转录聚合酶链反应法(RT-PCR)和计算机图像分析方法,观察其TRAF6基因的表达变化。结果TRAF6在小鼠成牙本质细胞系MDPC-23中呈阳性表达,当LPS干预后,TRAF6基因表达呈现上调趋势。结论本实验表明TRAF6参与调控LPS引发的成牙本质细胞炎症损伤过程中的细胞因子网络,推测TRAF6可能影响龋病的形成和发展过程。     

大鼠脑出血灶周HSP70凋亡表达及GM_1对其变化的影响

目的:研究大鼠脑出血灶周边组织HSP70的表达机制,探讨GM1抗细胞凋亡作用及对HSP70表达的影响。方法:1Wistar大鼠96只,随机分为:对照组,脑出血组,神经节苷酯干预组。大脑立体定位法复制脑出血模型。2免疫组化染色法检测脑组织HSP70表达。3吉姆萨染色法检测神经细胞凋亡。结果:1脑出血周边组织6h见HSP70表达,在6~48h呈上升趋势。2脑出血周边组织6h见凋亡细胞,在6~48h呈上升趋势。3GM1干预后HSP70表达与脑出血组比较显著上升,而凋亡细胞数下降,差异均具有显著性(P<0.05)。4GM1干预组中,局部比腹腔各时间点HSP70表达阳性细胞数增高,而凋亡细胞数下降,差异具有显著性(P<0.05)。结论:1HSP70参与脑出血周边组织损伤应答反应。2GM1可增强脑出血急性期HSP70的表达,同时具有抗细胞凋亡作用。3GM1可改善脑出血预后,且局部用药优于腹腔给药。     

热休克蛋白70在内毒素血症大鼠心肌中的表达及已酮可可碱的影响

目的：探讨热休克蛋白70在内毒素血症大鼠心肌组织中的表达及已酮可可碱的影响。方法：采用直接注射内毒素的方法建立大鼠急性内毒素血症模型。18只Wistar大鼠随机分成3组，假手术组，内毒素组和PTX组。分别采用免疫组织化学法和免疫印迹法检测心肌组织HSP70蛋白。结果：内毒素组免疫组织化学法与免疫印迹法检测心肌组织HSP70蛋白水平与假手术组比较都明显增强（P〈0．01）；予PTX干预后，免疫组织化学法与免疫印迹法检测心肌组织HSP70蛋白水平与内毒素组比较也都明显增强（P〈0．01）。结论：内毒素血症大鼠心肌组织内的HSP70蛋白表达可明显增高，PTX干预可以进一步增高内毒素血症大鼠心肌组织内的HSP70蛋白表达，从而进一步加强对心肌的保护作用。     

Expression of a small heat shock protein 27 (HSP27) in mouse skin tumors induced by UVB-irradiation

We investigated the expression of heat shock protein 27 (HSP27) at intermediate stages of a cutaneous tumor induced by UVB-irradiation stress (290-380 nm, max. 312 nm) using an immunostaining method. After 15-20 weeks of chronic exposure to UVB irradiation at a dose of 2 kJ/m2, HSP27 was found in the upper cell layers of bowenoid multilayers of epidermis, in areas of the lesions where normal stratification seems to be conserved. After 25 weeks, HSP27 was weakly expressed in squamous cell carcinoma (SCC). The HSP27 distribution patterns during cutaneous tumor progression resemble that of cytokeratin 10, a differentiation marker in keratinocytes. In SCC, a low degree of HSP27 expression was detected in the well-differentiated carcinomatous areas, but not in the poorly differentiated areas. These results indicate that the level of HSP27 decreases significantly as epithelial carcinoma growth progresses upon UVB-exposure. The expression of HSP27 may be associated with the onset of skin keratinocyte differentiation, but not with progression of SCC.     

Heavy metals chromium and neodymium reduced phosphorylation level of heat shock protein 27 in human keratinocytes

Heavy metals may exert their acute and chronic effects on the human skin through stress signals. In the present study, 2DE-based proteomics was used to analyze the protein expression in human keratinocytes exposed to heavy metals, chromium and neodymium, and 10 proteins with altered expression were identified. Among these proteins, small heat shock protein 27 (HSP27) was up-regulated significantly and the up-regulation was validated by Western blot and immunofluorescence. In addition, the mRNA expression level of HSP27 markedly increased as detected by quantitative PCR. More interestingly, the ratio of phosphorylated HSP27 and total HSP27 significantly decreased in keratinocytes treated with the heavy metals. These findings suggested that heavy metals reduced the phosphorylation level of HSP27, and that the ratio of p-HSP27 and HSP27 may represent a potential marker or additional endpoint for the hazard assessment of skin irritation caused by chemical products.     

5-氟尿嘧啶对口腔鳞癌细胞株热休克蛋白27／70表达的影响

目的：为探讨热休克蛋白27和热休克蛋白70在口腔鳞状细胞癌化疗中的意义。本实验选用口腔鳞状细胞癌细胞株OSC-4作为研究对象t随机分组。实验组加100μg／mL 5-氟尿嘧啶。方法：用MTT法检测细胞存活率,流式细胞仪检测细胞凋亡,Western Blot检洲HSP27和HSP70的表达。结果：OSC-4细胞经5-Fu处理后,MTT和流式细胞仪显示5-Fu抑制OSC-4细胞的存活率（P〈0．05）．并存在时间和剂量依赖。Western Blot结果显示HSP27和HSP70在OSC-4中均有表达,在OSC-4细胞中5-Fu能诱导HSP27和HSP70的表达（P〈0．05）。HSP27和HSP70在不同口腔鳞状细胞癌细胞株中均有表达,表达有差异（P〈0．05）。结论：5-Fu能诱导OSC-4细胞的凋亡,并呈现时间与剂量依赖;5-Fu能上调HSP27和HSP70的表达。     

Thyroxine pretreatment increases basal myocardial heat-shock protein 27 expression and accelerates translocation and phosphorylation of this protein upon ischaemia

Thyroxine pretreatment increases the tolerance of the heart to ischaemia, and heat-shock protein 27 (HSP27) is considered to play an important role in cardioprotection. The present study investigated whether long-term thyroxine administration can induce changes in the expression, translocation and phosphorylation of HSP27 at baseline and upon ischaemic stress. L-Thyroxine (T(4)) was administered to Wistar rats (25 microg/100 g/day s.c.) for 2 weeks, while normal animals served as controls. Hearts from normal and thyroxine-treated rats were perfused in Langendorff mode and subjected to 10 or 20 min of zero-flow global ischaemia only or to 20 min of ischaemia followed by 45 min of reperfusion. Total and phospho-HSP27 expression were assessed at different times in the Triton-soluble (cytosol-membrane), S fraction, and the Triton-insoluble (cytoskeleton-nucleus) fraction, P fraction. Postischaemic recovery of left ventricular developed pressure at 45 min of reperfusion was expressed as % of the initial value. In hearts from thyroxine-treated animals, the levels of basal total HSP27 and phospho-HSP27 in the P fraction were significantly increased as compared to normal. In response to ischaemia, in hearts from thyroxine-treated rats, the levels of total HSP27 and phospho-HSP27 were found to be significantly increased in the P fraction at 10 and 20 min of ischaemia as compared to preischaemic values, whereas in normal hearts, the levels of total HSP27 and phospho-HSP27 were significantly increased at 20 min only. Postischaemic functional recovery was significantly greater in thyroxine-treated than in untreated hearts. In summary, long-term thyroxine pretreatment results in an increased basal expression and phosphorylation of HSP27 and in an earlier and sustained redistribution of HSP27 from the S to the P fraction in response to ischaemia. This effect might be of important therapeutic relevance.     

Riboflavin protects mice against liposaccharide-induced shock through expression of heat shock protein 25

Riboflavin (vitamin B2) is a water-soluble vitamin essential for normal cellular functions, growth and development. This study aimed to investigate the effects of vitamin B2 on the survival rate, and expressions of tissue heat shock protein 25 (HSP25) and heat shock factor 1 (HSF1) in mice undergoing lipopolysaccharide (LPS) induced shock. Mice were assigned to four groups, saline vehicle, LPS, LPS plus low dose of vitamin B2 (LB2) and LPS plus high dose of vitamin B2 (HB2). Vitamin B2 (1 and 10 mg/kg BW) was administered intraperitoneally at 2 and 0 h before the i.p. administration of LPS. At the end of the experiment, the survival rate monitored was 10, 20, 60, and 100% for LPS, LB2, HB2, and saline mice, respectively. HSP25 expressions in the heart and lung were significantly enhanced in a time-dependent manner in the HB2 mice as compared to the saline mice (p < 0.05), but not altered in the LB2 mice. In the HB2 mice, plasma riboflavin concentrations reached 300 nM at 6 h post LPS and returned to the 0 h level at 72 h. The results showed that high dose of riboflavin could decrease LPS-induced mortality through an increased expression of HSP25.     

Introduction of antisense oligonucleotides to heat shock protein 47 prevents pulmonary fibrosis in lipopolysaccharide-induced pneumopathy of the rat

Acute respiratory distress syndrome (ARDS) confers high morbidity, and in part due to pulmonary fibrosis. The 47-kDa heat shock protein 47 (HSP 47) is a collagen-specific molecular chaperone that has been shown to play a major role in the processing and secretion of procollagen. We examined the effect of antisense oligonucleotides against HSP 47 in Wistar rats with lipopolysaccharide (LPS)-induced pulmonary fibrosis. These rats expressed heat shock protein (HSP) 47 and collagen in response to LPS. The distribution of HSP 47 was similar to that of collagen, and all control rats displayed pulmonary fibrosis after intratracheal administration of 20 mg/kg LPS alone. Antisense oligonucleotides (100 nmol/kg dissolved in saline) were administered with the LPS among experimental subjects. Subsequent immunoblot analysis confirmed the inhibition of HSP 47 by the administration of antisense oligonucleotides. The oligonucleotides significantly improved pulmonary fibrosis among those rats administered LPS, but the oligonucletides themselves did not produce any significant changes in the behavior or histology of the lungs among control rats. These findings suggest that HSP 47 antisense oligonucleotides improve lung fibrosis among rats with LPS-induced pneumopathy.