共查询到20条相似文献,搜索用时 15 毫秒
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Ding X.Z. Tsokos G.C. Smallridge R.C. Kiang J.G. 《Molecular and cellular biochemistry》1997,177(1-2):145-152
Thermotolerant cells display attenuated heat shock protein 70 kD (HSP-70) gene expression and signal transduction such as intracellular Ca2+ concentration and inositol trisphosphate in response to sublethal heat. To further investigate the regulation of heat shock gene expression, we developed constructs containing human HSP-70 and HSF1 genes and transfected human epidermoid A-431 cells. These cells were chosen because skin cells are especially vulnerable to heat shock and other environmental stressors. We report that A431 cells can be successfully transfected with HSP-70 and HSF1 genes as shown by the elevated levels of respective message and protein. Overexpression of HSP-70 in cells transfected with HSP-70 gene led to a down-regulation of the HSF1 gene expression. Interestingly, transfection of cells with the HSF1 gene was not associated with increased expression of HSP-70. Exposure of HSF1 gene-transfected cells to heat resulted in a transient but significant increase in HSP-70 gene expression as compared to that found in vector-transfected cells, which was completely inhibited by treatment with staurosporine. In conclusion, we have demonstrated successful transfection of human A-431 cells with HSF1 and HSP-70 genes, where the regulation of their expression can be studied. (Mol Cell Biochem 167: 145-152, 1997) 相似文献
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WANG Xiaolin XU Cunshuan WANG Xiujie WANG Dongjie WANG Qingshang ZHANG Baochen 《中国科学C辑(英文版)》2006,49(5):500-512
The mammal’s high elevation (hypoxia) adaptation was studied by using the immunological and the molecular biological methods
to understand the significance of Hsp (hypoxia) adaptation in the organic high elevation, through the mammal heat shock response.
(1) From high elevation to low elevation (natural hypoxia): Westem blot and conventional RT-PCR and real-time fluorescence
quota PCR were adopted. Expression difference of heat shock protein of 70 (Hsp70) and natural expression of brain tissue of
Hsp70 gene was determined in the cardiac muscle tissue among the different elevation mammals (yak). (2)From low elevation to high
elevation (hypoxia induction): The mammals (domestic rabbits) from the low elevation were sent directly to the areas with
different high elevations like 2300, 3300 and 5000 m above sea level to be raised for a period of 3 weeks before being slaughtered
and the genetic inductive expression of the brain tissue of Hsp70 was determined with RT-PCR. The result indicated that all of the mammals at different elevations possessed their heat shock
response gene. Hsp70 of the high elevation mammal rose abruptly under stress and might be induced to come into being by high
elevation (hypoxia). The speedy synthesis of Hsp70 in the process of heat shock response is suitable to maintain the cells’
normal physiological functions under stress. The Hsp70 has its threshold value. The altitude of 5000 m above sea level is
the best condition for the heat shock response, and it starts to reduce when the altitude is over 6000 m above sea level.
The Hsp70 production quantity and the cell hypoxia bearing capacity have their direct ratio. 相似文献
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Xiaolin Wang Cunshuan Xu Xiujie Wang Dongjie Wang Qingshang Wang Baochen Zhang 《中国科学:生命科学英文版》2006,49(5):500-512
The mammal's high elevation(hypoxia) adaptation was studied by using the immu-nological and the molecular biological methods to understand the significance of Hsp(hypoxia) ad-aptation in the organic high elevation,through the mammal heat shock response.(1) From high ele-vation to low elevation(natural hypoxia) :Western blot and conventional RT-PCR and real-time fluo-rescence quota PCR were adopted.Expression difference of heat shock protein of 70(Hsp70) and natural expression of brain tissue of Hsp70 gene was determined in the cardiac muscle tissue among the different elevation mammals(yak) .(2) From low elevation to high elevation(hypoxia induction) :The mammals(domestic rabbits) from the low elevation were sent directly to the areas with different high elevations like 2300,3300 and 5000 m above sea level to be raised for a period of 3 weeks be-fore being slaughtered and the genetic inductive expression of the brain tissue of Hsp70 was deter-mined with RT-PCR.The result indicated that all of the mammals at different elevations possessed their heat shock response gene.Hsp70 of the high elevation mammal rose abruptly under stress and might be induced to come into being by high elevation(hypoxia) .The speedy synthesis of Hsp70 in the process of heat shock response is suitable to maintain the cells' normal physiological functions under stress.The Hsp70 has its threshold value.The altitude of 5000 m above sea level is the best condition for the heat shock response,and it starts to reduce when the altitude is over 6000 m above sea level.The Hsp70 production quantity and the cell hypoxia bearing capacity have their direct ratio. 相似文献
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Heat shock proteins of chicken lens 总被引:12,自引:0,他引:12
The presence of heat shock proteins HSP-40, HSP-70, and HSc-70 in adult and embryonic chicken lenses were determined. The epithelium, cortex, and nucleus of adult chicken lens were separated and tested for the presence of heat shock proteins (hsps) by western blot, using specific antibodies for HSP-40, HSP-70, and HSc-70. Water soluble (WSF) and water insoluble fractions (WIF) of embryonic chicken lenses were isolated and tested for the presence of HSP-40, HSP-70, and HSc-70 by immunoblot. Embryonic chicken lens sections were also analyzed for the presence of heat shock proteins by immunofluorescence technique. Data obtained from these experiments revealed that HSP-40, HSP-70, and HSc-70 are present in all areas of both adult and embryonic chicken lens. Presence of hsps protein in the deep cortex and nucleus is intriguing as no detectable metabolic activities are reported in this area. However it can be proposed that hsps HSP-40, HSP-70, and HSc-70 can interact with protein of these areas and protect them from stress induced denaturation. 相似文献
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Heat shock proteins of adult and embryonic human ocular lenses. 总被引:11,自引:0,他引:11
We investigated the presence and distribution of heat shock proteins, HSP-70 [Horwitz, J. 1992. Proc Natl Acad Sci 89:10449-10453], HSP-40, HSc-70, HSP-27, and alphabeta-crystallin in different regions of adult and fetal human lenses and in aging human lens epithelial cells. This study was undertaken because heat shock proteins may play an important role in the maintenance of the supramolecular organization of the lens proteins. Human adult and fetal lenses were dissected to separate the epithelium, superficial cortex, intermediate cortex, and nucleus. The water soluble and insoluble protein fractions were separated by SDS-PAGE, and transferred to nitrocellulose paper. Specific antibodies were used to identify the presence of heat shock proteins in distinct regions of the lens. HSP-70 [Horwitz, 1992], HSP-40, and HSc-70 immunoreactivity was mainly detected in the epithelium and superficial cortical fiber cells of the adult human lens. The small heat shock proteins, HSP-27 and alphabeta-crystallin were found in all regions of the lens. Fetal human lenses showed immunoreactivity to all heat shock proteins. An aging study revealed a decrease in heat shock protein levels, except for HSP-27. The presence of HSP-70 [Horwitz, 1992], HSP-40, and HSc-70 in the epithelium and superficial cortical fiber cells imply a regional cell specific function, whereas the decrease of heat shock protein with age could be responsible for the loss of optimal protein organization, and the eventual appearance of age-related cataract. 相似文献
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Cell signaling and heat shock protein expression 总被引:5,自引:0,他引:5
Exposure of cells and organs to heat shock is associated with numerous changes in various cellular metabolic parameters and overexpression of proteins collectively known as heat shock proteins (HSP). In this communication we review the cell-signaling events that are altered in response to heat shock as they relate to the subsequent induction of HSP 70 kd (HSP-70) expression. We also review the mechanisms by which HSP-70 is involved in conferring cytoprotective effects. The possibility of altering HSP expression through manipulations of the cell-signal process has clinical importance.The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or reflecting the views of the Department of the Army or Department of Defense. 相似文献
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Kiang JG Kiang SC Juang YT Tsokos GC 《American journal of physiology. Gastrointestinal and liver physiology》2002,282(3):G415-G423
The nitric oxide (NO) synthase inhibitor N(omega)-nitro-L-arginine (L-NNA) inhibits heat stress (HS)-induced NO production and the inducible 70-kDa heat shock protein (HSP-70i) in many rodent organs. We used human intestinal epithelial T84 cells to characterize the inhibitory effect of L-NNA on HS-induced HSP-70i expression. Intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured using fura-2, and protein kinase C (PKC), and PKA activities were determined. HS increased HSP-70i mRNA and protein in T84 cells exposed to 45 degrees C for 10 min and allowed to recover for 6 h. L-NNA treatment for 1 h before HS inhibited the induction of HSP-70i mRNA and protein, with an IC(50) of 0.0471 +/- 0.0007 microM. Because the HS-induced increase in HSP-70i mRNA and protein is Ca(2+) dependent, we measured [Ca(2+)](i) after treating cells with L-NNA. L-NNA at 100 microM significantly decreased resting [Ca(2+)](i). Likewise, treatment with 1 microM GF-109203X or H-89 (inhibitors of PKC and PKA, respectively) for 30 min also significantly decreased [Ca(2+)](i) and inhibited HS-induced increase in HSP-70i. GF-109203X- or H-89-treated cells failed to respond to L-NNA by further decreasing [Ca(2+)](i) and HSP-70i. L-NNA effectively blocked heat shock factor-1 (HSF1) translocation from the cytosol to the nucleus, a process requiring PKC phosphorylation. These results suggest that L-NNA inhibits HSP-70i by reducing [Ca(2+)](i) and decreasing PKC and PKA activity, thereby blocking HSF1 translocation from the cytosol to the nucleus. 相似文献
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Genistein inhibits herbimycin A-induced overexpression of inducible heat shock protein 70 kDa 总被引:1,自引:0,他引:1
Kiang JG 《Molecular and cellular biochemistry》2003,245(1-2):191-199
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Katada K Naito Y Shimozawa M Mizushima K Kuroda M Takagi T Kokura S Ichikawa H Yoshida N Matsui H Yoshikawa T 《Redox report : communications in free radical research》2004,9(6):337-342
Recent investigations have demonstrated that the signaling of hypoxia-re-oxygenation is a major contributing pathway leading to gastric mucosal injury induced by stress, non-steroidal anti-inflammatory drugs, and Helicobacter pylori. The aim of the present study was to perform a gene expression analysis on the gastric mucosal cellular response to hypoxia-reoxygenation using a high-density oligonucleotide array. Cells were subjected to hypoxia with 95% N(2) and 5% CO(2) at 37 degrees C for 2 h. Reoxygenation was initiated by placing the cells in an environment of normoxia for 2 h. Total RNA was extracted, and differences in gene expression profiles between the normoxia and hypoxia-reoxygenation groups were investigated using a GeneChip of Rat Toxicology U34 array (Affymetrix). Hypoxia-reoxygenation up-regulated the stress-related genes (heat shock protein-70 [HSP-70], catalase). The enhanced expression of HSP-70 was confirmed by Western blot analysis. In conclusion, these results suggest that up-regulation of the HSP-70 gene after reoxygenation may play a role in maintaining cell survival and supporting cell function as a molecular chaperone. 相似文献
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Cerebellar granule cells (CGC) die apoptotically after five days in culture (DIV) at physiological concentrations of potassium (5 mM; K5). When CGC are depolarized (K25) or treated with NMDA (150 M) cell survival is increased. CGC changed from K25 to K5 die after 24–48 h. It is known that heat shock protein (HSP) may protect from cell death. Here, we found that cells in K5 showed an increase in HSP-70 levels after 3 DIV. Similarly, in cells changed from K25 to K5, HSP-70 levels were increased after 6 h. Neither NMDA nor K25 treatment affected HSP-70 levels from 2–7 DIV. Ethanol or thermal stress induced HSP-70, but cell survival was not affected in K5 medium. These results suggest that HSP, particularly HSP-70, are not involved in the mechanisms by which NMDA and KCl promote cell survival. 相似文献
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Glutamine's protection against cellular injury is dependent on heat shock factor-1 总被引:12,自引:0,他引:12
Morrison AL Dinges M Singleton KD Odoms K Wong HR Wischmeyer PE 《American journal of physiology. Cell physiology》2006,290(6):C1625-C1632
Glutamine (GLN) has been shown to protect cells, tissues, and whole organisms from stress and injury. Enhanced expression of heat shock protein (HSP) has been hypothesized to be responsible for this protection. To date, there are no clear mechanistic data confirming this relationship. This study tested the hypothesis that GLN-mediated activation of the HSP pathway via heat shock factor-1 (HSF-1) is responsible for cellular protection. Wild-type HSF-1 (HSF-1+/+) and knockout (HSF-1/) mouse fibroblasts were used in all experiments. Cells were treated with GLN concentrations ranging from 0 to 16 mM and exposed to heat stress injury in a concurrent treatment model. Cell viability was assayed with phenazine methosulfate plus tetrazolium salt, HSP-70, HSP-25, and nuclear HSF-1 expression via Western blot analysis, and HSF-1/heat shock element (HSE) binding via EMSA. GLN significantly attenuated heat-stress induced cell death in HSF-1+/+ cells in a dose-dependent manner; however, the survival benefit of GLN was lost in HSF-1/ cells. GLN led to a dose-dependent increase in HSP-70 and HSP-25 expression after heat stress. No inducible HSP expression was observed in HSF-1/ cells. GLN increased unphosphorylated HSF-1 in the nucleus before heat stress. This was accompanied by a GLN-mediated increase in HSF-1/HSE binding and nuclear content of phosphorylated HSF-1 after heat stress. This is the first demonstration that GLN-mediated cellular protection after heat-stress injury is related to HSF-1 expression and cellular capacity to activate an HSP response. Furthermore, the mechanism of GLN-mediated protection against injury appears to involve an increase in nuclear HSF-1 content before stress and increased HSF-1 promoter binding and phosphorylation. knockout cells; amino acid; heat stress mechanism 相似文献
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Inducible heat shock protein 70 kD and inducible nitric oxide synthase in hemorrhage/resuscitation-induced injury 总被引:1,自引:0,他引:1
Kiang JG 《Cell research》2004,14(6):450-459
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Heat shock of rabbit synovial fibroblasts increases expression of mRNAs for two metalloproteinases, collagenase and stromelysin 总被引:2,自引:0,他引:2
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Two metalloproteinases, collagenase and stromelysin, are produced in large quantities by synovial fibroblasts in individuals with rheumatoid arthritis. These enzymes play a major role in the extensive destruction of connective tissue seen in this disease. In this study, we show that heat shock of monolayer cultures of rabbit synovial fibroblasts increases expression of mRNA for heat shock protein 70 (HSP-70), and for collagenase and stromelysin. We found that after heat shock for 1 h at 45 degrees C, the mRNA expression for HSP-70 peaks at 1 h and returns to control levels by 3 h. Collagenase and stromelysin mRNA expression is coordinate, reaching peak levels at 3 h and returning to control levels by 10 h. The increase in mRNA is paralleled by an increase in the corresponding protein in the culture medium. 3 h of heat shock at a lower temperature (42 degrees C) is also effective in inducing collagenase and stromelysin mRNAs. Concomitant treatment with phorbol myristate acetate (PMA; 10(-8) or 10(-9) M) and heat shock is not additive or synergistic. In addition, all-trans-retinoic acid, added just before heat shock, prevents the increase in mRNAs for collagenase and stromelysin. Our data suggest that heat shock may be an additional mechanism whereby collagenase and stromelysin are increased during rheumatoid arthritis and perhaps in other chronic inflammatory stress conditions. 相似文献