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1.
目的 了解Ad-热休克蛋白70(HSP70)对缺氧/复氧损伤后肠上皮细胞线粒体功能及能量代谢的影响.方法 取肠上皮细胞株IEC-6分别转染Ad-HSP70腺病毒载体和空腺病毒载体,蛋白质印迹法观察HSP70的表达.取IEC-6细胞分为正常对照组(不作任何处理)、缺氧/复氧组(给予缺氧/复氧处理)、Ad-HSP70转染组(转染Ad-HSP70腺病毒载体后,给予缺氧/复氧处理).采用噻唑蓝比色法检测细胞内线粒体脱氧酶的活性,高效液相色谱分析法测定细胞能量代谢.结果 与转染空腺病毒载体相比,转染Ad-HSP70腺病毒载体可显著增加细胞HSP70的表达.缺氧/复氧组细胞内线粒体脱氢酶的活性明显低于正常对照组(P<0.01),Ad-HSP70转染组该指标明显高于缺氧/复氧组(P<0.01).缺氧/复氧组细胞内腺苷三磷酸含量较正常对照组显著下降,而腺苷二磷酸、腺苷-磷酸含量显著升高;Ad-HSP70转染组细胞能量指标与正常对照组相似(P>0.05),较缺氧/复氧组有明显改善(P<0.05或P<0.01).缺氧/复氧组细胞能荷为0.615±0.060,明显低于正常对照组(0.748±0.012,P<0.01)、Ad-HSP70转染组(0.736±0.028,P<0.01).结论 Ad-HSP70腺病毒载体转染肠上皮细胞可诱导HSP70表达增加,显著提高细胞缺氧/复氧后胞内腺苷三磷酸的含量及细胞能荷,保护线粒体整体功能,提示线粒体是HSP70保护肠上皮细胞缺氧/复氧损伤的主要靶细胞器之一.  相似文献   

2.
Insulin potentiates expression of myocardial heat shock protein 70.   总被引:7,自引:0,他引:7  
OBJECTIVE: Since insulin stimulates nitric oxide (NO) production and an increase in NO following heat shock is required for myocardial heat shock protein 70 (Hsp70) synthesis, we hypothesized that insulin would enhance myocardial Hsp70 synthesis by augmenting NO signaling. We examined whether a physiologic dose of insulin increased myocardial Hsp70 in unstressed and heat shock treated rats. METHODS: Adult male Sprague-Dawley rats were assigned to groups: (1) control, (2) insulin injected (200 microU/gm body weight), (3) heat shock treated (core body temperature 42 degrees C for 15 min), (4) heat shock and insulin treated, (5) L-nitroarginine methyl ester (L-NAME) and heat shock and insulin treated, (6) sodium nitroprusside (SNP) and heat shock and insulin treated. Six hours later, myocardial Hsp70 content and localization was analyzed. RESULTS: Hsp70 was increased in heat shock treated hearts (120.6+/-16.8 ng/mg protein, P < 0.001) vs. control (12.9+/-2.0 ng/mg protein), or insulin treated hearts (15.5+/-0.83 ng/mg protein). In addition, Hsp70 was increased in the heat shock and insulin treated hearts (164.4+/-7.53 ng/mg protein) compared to control, insulin only (P = 0.001) or heat shock only treated hearts (P = 0.01). L-NAME did not abolish the insulin induced increase in Hsp70 in heat shocked hearts (195.2+/-13.4 ng/mg protein, P = 0.21) and SNP did not further enhance Hsp70 in the insulin and heat shocked group (188.9+/-8.2 ng/mg protein, P = 0.71). Western analysis and confocal microscopy revealed a lowlevel expression of myocardial Hsp70 in response to insulin. Hsp70 was localized primarily in blood vessels after insulin or heat shock treatments. CONCLUSIONS: Insulin caused a low-level expression of myocardial Hsp70 and potentiated Hsp70 synthesis in response to heat shock. The ability of insulin to potentiate Hsp70 after heat shock is independent of NO signaling as it was not altered by either LNAME or SNP pretreatment. Blood vessels appear to be the primary site of Hsp70 after insulin or heat shock treatment.  相似文献   

3.
目的 探讨热休克蛋白70(HSP 70)与雄激素非依赖性前列腺癌(PC a)发生的关系。方法 采用免疫组化方法测定2例正常前列腺组织、20例雄激素依赖性PCa和14例雄激素非依赖性PCa石蜡包埋标本HSP 70的表达。结果 HSP 70在正常前列腺基底细胞中呈阳性表达,分泌细胞中为阴性表达。20例雄激素依赖性PCa中16例(80.0%)无HSP 70表达,3例有微弱表达,1例为异质性表达。14例雄激素非依赖性PCa中13例(92.8%)呈弥漫、高水平的HSP 70表达,1例为阴性,与雄激素依赖PCa HSP70表达率的比较,差别有极显著性意义(P<0.01)。结论 HSP 70过表达与雄激素依赖性PCa向雄激素非依赖性PCa的进展有关。  相似文献   

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6.
An analysis of postoperative complications and survival was conducted in 31 patients undergoing pancreatoduodenectomy (PD) for carcinoma of the pancreas or periampullary carcinoma. Of them, 11 were over 70 years of age and 20 were under 70. Anastomotic leakage was the most common complication after PD. Definite pancreatic leakage was found in one patient in the over 70 group, and one case each of pancreatic, biliary, and gastric leakage were found in the under 70 group. All complications were treated conservatively without any further operative intervention. The overall morbidity rate was 41.9% (13/31), being 45.5% (5/11) in the over 70 group and 40.0% (8/20) in the under 70 group, and no operative deaths occurred within 30 days after surgery. The cumulative survival rate of the patients aged over 70 years with carcinoma of the pancreas or periampullary carcinoma did not differ significantly from the rate of those under 70. It was thus concluded that PD achieves an adequate prognosis and survival in patients over 70 years of age.  相似文献   

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8.
The objective of this study is to investigate the effects of mild electrical stimulation (MES) and heat stress (HS) on heat shock protein 70 (HSP70), that protects chondrocytes and enhances cartilage matrix metabolism, in chondrocyte and articular cartilage. Rabbit articular chondrocytes were treated with MES and/or HS. The safeness was assessed by LDH assay and morphology. HSP70 protein, ubiquitinated proteins and HSP70 mRNA were examined by Western blotting and real‐time PCR. Rat knee joints were treated with MES and/or HS. HSP70 protein, ubiquitinated proteins, HSP70 mRNA and proteoglycan core protein (PG) mRNA in articular cartilage were investigated. In vitro, HS increased HSP70 mRNA and HSP70 protein. MES augmented ubiquitinated protein and HSP70 protein, but not HSP70 mRNA. MES + HS raised HSP70 mRNA and ubiquitinated protein, and significantly increased HSP70 protein. In vivo, HS and MES + HS treatment augmented HSP70 mRNA. HS modestly augmented HSP70 protein. MES + HS significantly increased HSP70 protein and ubiquitinated proteins. PG mRNA was markedly raised by MES + HS. This study demonstrated that MES, in combination with HS, increases HSP70 protein in chondrocytes and articular cartilage, and promotes cartilage matrix metabolism in articular cartilage. MES in combination with HS can be a novel physical therapy for osteoarthritis by inducing HSP70 in articular cartilage. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 894–900, 2013  相似文献   

9.
BACKGROUND: We recently demonstrated that heat stress induction of heat shock protein 70 (HSP70) in donor animals before harvest decreases posttransplant ischemia-reperfusion injury in preserved rat lung isografts. The purpose of this study was to investigate the feasibility of HSP70 gene transfection into rat lung isografts using an adenoviral vector, and to study the effects of gene expression on subsequent ischemia-reperfusion injury. METHODS: In preliminary studies to determine the optimal titer, animals were injected with various titers of adenovirus-HSP70 (saline, 5 x 10(9), 1 x 10(10), and 2 x 10(10) plaque forming units [pfu]) and sacrificed 5 days after injection. To determine the optimal exposure time, animals were sacrificed at different times (0, 6, 24, and 72 hours) after intravenous injection of adenovirus-HSP70. In a subsequent series of transplant experiments, donors were allocated to three groups according to transfection strategy. Group 1 (n = 8) donors received 5 x 10(9) pfu adenovirus-HSP70 intravenously, group 2 (n = 7) donors received 5 x 10(9) pfu adenovirus-beta-galactosidase (as a virus control), and group 3 (n = 7) donors received saline and served as a negative control. Twenty-four hours after treatment all grafts were harvested and stored for 18 hours before orthotopic left lung transplantation. Twenty-four hours after implantation animals were sacrificed for assessment. The expression of HSP70 was assessed by Western blot analysis. RESULTS: In preliminary studies, HSP70 was detectable even at low titers (5 x 10(9) pfu) of adenovirus-HSP70, and was detectable at low levels as early as 6 hours after intravenous administration. Heat shock protein 70 expression was maximal at 24 hours. In transplant experiments, Western blot analysis showed that overexpression of HSP70 occurred in the HSP70-transfected lungs. The mean arterial oxygenation 24 hours after reperfusion in group 1 was superior in comparison with other groups (p < 0.05). Wet to dry weight ratio (p < 0.05) and myeloperoxidase activity (p < 0.05) were also significantly less in group 1 grafts compared with the other groups. CONCLUSIONS: This study demonstrates that in vivo, donor adenovirus-mediated gene transfer of HSP70 decreases subsequent ischemia-reperfusion injury in rat lung isografts.  相似文献   

10.
热休克蛋白70 mRNA在膀胱癌中的表达及与细胞凋亡的关系   总被引:4,自引:0,他引:4  
目的 探讨热休克蛋白 70 (HSP 70 )mRNA在膀胱移行细胞癌中的表达及与细胞凋亡的关系。 方法 应用原位杂交法对 6 0例膀胱癌组织中HSP 70mRNA进行检测 ,原位DNA末端标记法 (TUNEL)检测细胞凋亡情况。 结果  6 0例标本中 ,HSP 70mRNA阳性表达率为 5 6 .7% (34/ 6 0 )。HSP 70mRNA表达随肿瘤分级增高而增加 (P <0 .0 5 ) ;G1,G2 和G3 各级间两两比较 ,HSP 70mRNA表达差异也有显著性意义 (P <0 .0 5 )。随着肿瘤分期增高 ,HSP 70mRNA表达随之增加 (P <0 .0 5 ) ;Ⅰ ,Ⅱ和Ⅲ各期间两两比较 ,差异有显著性意义 (P <0 .0 5 )。肿瘤细胞凋亡指数 (AIs)随恶性程度的增高显著降低 (P <0 .0 5 ) ;HSP 70mRNA表达率和AIs呈负相关 (rs=- 0 .6 85 ,P <0 .0 5 )。 结论 HSP 70mRNA表达随膀胱癌恶性程度增加显著增高 ,细胞凋亡则显著降低 ,提示HSP 70表达能够抑制膀胱癌细胞的凋亡。  相似文献   

11.
目的:探讨热休克蛋白70(HSP70)敲低对结肠癌细胞在裸鼠体内生长的影响。方法:构建两种干扰HSP70的shRNA(HSP70 shRNA-1和HSP70 shRNA-2)质粒表达载体,分别转染到结肠癌HT29细胞,并以空质粒转染(阴性对照)和未转染的HT29细胞(空白对照)为对照,用RT-PCR和Western blot方法检测各组HT29细胞HSP70基因和蛋白的表达;将HSP70shRNA-2转染、空质粒转染及未转染的HT29细胞分别接种至裸鼠皮下建立移3组植瘤模型,观察肿瘤生长情况,3周后剥离瘤块,用HE染色、免疫组化和TUNEL法分别检测移植瘤组织形态学、增殖细胞核抗原(PCNA)的表达及细胞凋亡情况。结果:RT-PCR和Western blot结果显示HSP70 shRNA-1和HSP70 shRNA-2均能明显抑制HT29细胞HSP70基因和蛋白的表达,且HSP70 shRNA-2的抑制作用更为明显,空质粒转染对HSP70的表达无明显影响;与空白对照组比较,HSP70 shRNA-2转染组裸鼠移植瘤生长明显较明显减慢(P<0.01),瘤组织中心出现明显坏死,PCNA表达明显降低,细胞凋亡率明显增加(P<0.01),而空质粒转染组无明显上述改变(P>0.05)。结论:HSP70沉默能抑制结肠癌HT29细胞裸鼠移植瘤的生长,促进细胞凋亡,HSP70可能是治疗结肠癌有效靶点。  相似文献   

12.
目的:比较急性胆囊炎患者腹腔镜手术前后血液中热休克蛋白70(HSP70)的表达差异,从分子和蛋白质水平探讨HSP70与急性胆囊炎的关系。方法:以急性胆囊炎行腹腔镜手术的住院患者85例为研究对象,应用酶联免疫吸附法(ELISA)检测手术前后的静脉血中HSP70的量;随机抽取其中4例,提取其血液中的总RNA和蛋白质,分别用RT-PCR和Western-blotting检验HSP70在mRNA和蛋白质水平的表达,并用Bio-rad公司的quantity one软件进行图像数据分析。结果:ELISA结果显示,急性胆囊炎患者腹腔镜手术前HSP70为181.42±13.58,手术后为63.74±9.03,差异有显著性(P<0.05)。RT-PCR和Western-blotting结果显示,患者腹腔镜手术后HSP70水平较手术前显著降低。结论:HSP70与急性胆囊炎关系密切,可能系因急性炎症刺激机体的HSP70表达增高,而腹腔镜手术清除炎症病灶后HSP70的表达迅速降低。  相似文献   

13.
OBJECTIVE: Proctocolectomy and ileal pouch anal anastomosis (IPAA) has become the standard surgery for patients with mucosal ulcerative colitis (MUC). Although there is no absolute age limitation, there are concerns as to its use in elderly patients due to the risks of potential complications and poor function. The aim of this study was to assess the complications and outcome of patients over the age of 70 years with MUC who underwent IPAA. Results in these patients were compared to the results in a group of patients aged less than 70 years who had IPAA. METHODS: After Institutional Review Board approval, a retrospective review of the medical records of patients with MUC who underwent IPAA was undertaken. These patients were divided into four age groups: <30 years of age, 30-49 years, 50-69 years, >or=70 years. RESULTS: From 1989 to 2001, 330 patients underwent IPAA for preoperative clinical and histopathological and postoperative histopathologically confirmed MUC; 17 were aged>or=70 years. The mean hospital stay was 5.8 (SEM 0.7) days in the patients aged<70 years and 6.0 (SEM 0.4) days in the patients aged>or=70 years (P=0.911). Postoperative complications occurred in 39% of patients>or=70 years and in 40% in the <70 years group (P=0.08). Pouch failure occurred in two (11.8%) patients>or=70 years and in 6 (1.9%)<70 (P=0.2). CONCLUSION: IPAA is a safe and feasible option in MUC patients over the age of 70 with functional results similar to results seen in younger patients.  相似文献   

14.
目的 探讨血液透析对不同年龄段尿毒症患者血清热休克蛋白70的影响.方法 对2010年4月在我院进行维持性血液透析的尿毒症患者,根据年龄将92例患者分为3组(青年组、中年组、老年组).在透析前、后分别采用酶联免疫复合物法测定血清热休克蛋白70水平.结果 青年组透析前、后热休克蛋白70水平比较差异有统计学意义(P<0.05...  相似文献   

15.
The Diagnostic Related Group (DRG) hospital payment system may be inequitable for certain groups of Medicare patients. This study of 216 Medicare urology patients in the ten non-age stratified urology DRGs demonstrated that patients seventy years of age and older (70+) had higher resource consumption than patients under seventy years of age (70-). Findings were: (1) older patients (70+) had higher total hospital costs (+12,022 per patient) than younger patients (70-) (+9,872 per patient); (2) a longer hospital length of stay (14.2 days vs 11.6 days); (3) financial risk of +1,756 loss per (70+) patient vs +1,309 profit per (70-) patient (p less than 0.05); (4) more diagnoses and procedures per patient, and (5) a higher mortality (4.0% vs 3.3%). These findings suggest that the current DRG scheme may be inequitable vis-a-vis the older urology patient in non-age stratified DRGs, and thus could limit access and quality of care for these patients in the future.  相似文献   

16.
BACKGROUND: Heat shock protein 70 (HSP70) is induced by a wide variety of stresses in addition to hyperthermia. Recent studies have clarified that mechanical stretching and pressure overload can induce HSP70 in some tissues and cells. However, it remains unclear whether HSP70 is induced in stretch-subjected lungs, such as those under mechanical ventilation. This study was designed to investigate the effects of high peak airway pressure (PAP) ventilation on HSP70 expression in intact rat lungs. METHODS: Male Sprague-Dawley rats were randomly allocated to one of three groups: non-ventilated (anesthesia alone) control group; PAP 15 cm H(2)O group (P15); and PAP 30 cm H(2)O group (P30). The rats in the PAP groups were subjected to pressure-controlled assisted ventilation at the appropriate PAP for 30 min. Rats were killed at 12, 24 and 48 h after ventilation or anesthesia alone, and the lungs were removed. The lung tissues were processed for immunohistochemical and Western blotting analyses of HSP70. RESULTS: Following 30 min of pressure-controlled assisted ventilation, HSP70 expression in the P30 group was significantly up-regulated in bronchiolar cells and subepithelial tissues at 12 h, and this up-regulation continued throughout the observation period. In contrast, there were no significant differences between the control and P15 groups, although the expression of HSP70 was higher in the P15 group than in the control group at all time points. CONCLUSIONS: HSP70 was induced by high PAP ventilation, but its specific role and induction mechanism remain unclear. Therefore, further investigations should be encouraged.  相似文献   

17.
热休克蛋白70对肾脏缺血预处理保护作用的研究   总被引:3,自引:0,他引:3  
目的 :研究缺血预处理对肾脏缺血再灌注损伤的保护作用 ,观察热休克蛋白 70 (HSP70 )的表达变化并探讨其作用机制。方法 :建立大鼠肾脏缺血再灌注损伤模型并进行缺血预处理 ,实验分组 :假手术组 (S组 )、缺血再灌注组 (IR组 )和缺血预处理组 (PC组 )。各组再灌注后检测血清肌酐 (Scr)、肾组织丙二醛 (MDA)含量 ,肾组织石蜡切片苏木精伊红染色以及免疫组化染色。结果 :PC组Scr值、肾小管病理评分、肾组织中MDA含量明显低于IR组 (P <0 .0 5 ) ,PC组与S组比较差异无显著性意义 (P >0 .0 5 ) ;HSP70免疫组化染色 :S组未见明显的阳性反应产物 ,PC组和IR组肾小管上皮细胞胞质可见棕黄色阳性反应产物。计算机图像分析显示PC组灰度值显著高于IR组 (P <0 .0 1)。结论 :缺血预处理对肾脏缺血再灌注损伤有明显保护作用 ,其作用机制可能与HSP70本身的细胞保护作用及HSP70的细胞内抗氧化作用有关  相似文献   

18.
BACKGROUND: Although extracellular heat-shock protein 70 (Hsp70) potentially mediates an inflammatory response, the association of circulating Hsp70 with complications after surgery is poorly understood. METHODS: Perioperative plasma concentrations of Hsp70 and interleukin-6 were measured by immunoassays in 64 consecutive patients undergoing liver resection. RESULTS: Plasma concentrations of Hsp70 and interleukin-6 showed a striking increase immediately after surgery, and on postoperative day 1. The Hsp70 levels correlated significantly with operation time, hepatic ischemia time, postoperative alanine aminotransferase levels, and maximum interleukin-6 levels (P <0.01). The Hsp70 and interleukin-6 concentrations were associated significantly with postoperative infection (P <0.05); Hsp70 concentrations and blood loss but not interleukin-6 were associated significantly with postoperative organ dysfunction (P <0.05) in multivariate analyses. CONCLUSIONS: These results suggest that circulating Hsp70 and IL-6 potentially play a pivotal role in pathophysiology of postoperative infection, and that circulating Hsp70 and blood loss may represent a prognostic marker for postoperative organ dysfunction.  相似文献   

19.
目的 探讨热休克蛋白(HSP)70-2基因沉默对精原细胞的影响及其机制.方法 构建HSP70-2特异的短发夹RNA(shRNA)质粒载体,并转染大鼠精原细胞;应用逆转录-聚合酶链反应(RT-PCR)检测HSP70-2 mRNA表达,Western blot检测HSP70-2、Cyclin DI、bcl-2和box蛋白表达,应用流式细胞术分析细胞周期和凋亡.结果 HSP70-2基因沉默组的HSP70-2蛋白表达量为0.93±0.13,明显低于阴性对照组1.31±0.10(P<0.01);流式细胞术检测结果显示,与转染Scrambled的阴性对照组比较,转染pGenesil-1-HSP70-2重组质粒组的PC3细胞在G1期出现明显细胞周期阻滞和细胞凋亡,G1期阻滞细胞比例为(80.09±2.37)%,凋亡率为(7.44±2.41)%,而阴性对照组分别为(61.85±2.31)%和(1.09±0.57)%(P<0.05).与阴性对照组比较,转染后细胞bcl-2、Cyclin D1表达水平显著降低,而bax表达水平升高.结论 HSP70-2基因沉默能诱导精原细胞凋亡,其机制可能是通过Cyclin D1蛋白调节细胞周期并通过上调box蛋白表达促进细胞凋亡.  相似文献   

20.
BACKGROUND: The heat shock proteins are increasingly becoming associated with immunopathologic phenomena, being induced in response to inflammation. They are highly immunogenic and are postulated as playing a role in both innate and adaptive immunity. Their proposed role in peptide binding and antigen presentation could suggest a potential role in the alloreactive process that leads to graft-versus-host disease (GVHD) after bone marrow transplantation. METHODS: In this study we examined the expression of the widely studied heat shock protein 70 (hsp70) in an in vitro-generated graft-versus-host reaction in human skin, using streptavidin biotin immunohistochemistry and laser scanning confocal microscopy. RESULTS: Hsp70 expression was correlated with high graft-versus-host responses (P<0.001) and was confirmed using laser scanning confocal microscopy. Increased expression of hsp70 was further defined due to increases in the inducible form of hsp70. Expression of inducible hsp70 was predictive of both clinical acute GVHD (P=0.001) and incidence of chronic GVHD (P<0.001). CONCLUSIONS: This investigation has demonstrated for the first time the expression of hsp70 in a human model of GVHD, suggesting involvement in the pathogenesis of the disease and providing the basis for further investigation. Increased expression of inducible hsp70 in the model could provide a biologic marker for the prediction of clinical acute and chronic GVHD.  相似文献   

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