• Title/Summary/Keyword: heat shock protein 70

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Expression of HSP70 Immunoreactivity in EPO Treated Rat Kidney (콩팥에서 Erythropoietin 투여로 인한 HSP70의 발현 변화)

  • Jung, Ju-Young;Kim, Jin
    • Applied Microscopy
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    • v.37 no.3
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    • pp.167-174
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    • 2007
  • Heat shock protein (HSP) 70 functions as a molecular chaperon and reduces stress-induced denaturation and aggregation of intracellular proteins. Erythropoietin (EPO) plays an important role during acute renal failure repair process by rapidly correcting anemia and enhancing renal tubular regeneration. The purpose of this study was to examine the effect of EPO treatment on renal HSP70 expression. Male Sprague-Dawley rats were injected rHUEPO. Kidney were preserved by in vivo perfusion with paraformaldehyde-lysine-periodate (PLP) and processed for immunohistochemistry and electron microscopy. In control kidney, HSP70 was expressed in the cortex, outer medulla and inner medulla. Especially, HSP immunoreactiviy was mainly founded in descending thin limb of outer medulla and inner medullary collecting duct. In EPO treated kidney, HSP70 expression markedly increased in the descending thin limb of outer medulla and newly detected in cortical collecting duct. Electron microscopy showed the presence of HSP immunoreactivity on the intracelluar vesicles and Golgi complex of descending thin limb and cortical collecting duct. These findings suggest that EPO treatment leads to new production of HSP70 in renal tubular cells, and induction of HSP70 by rHuEPO is causally related to protective function.

Expression and Localization of Heat Shock Protein 70 in Frozen-thawed IVF and Nuclear Transferred Bovine Embryos

  • Chung, K.S.;Choi, Y.J.;Song, S.J.;Do, J.T.;Yoon, B.S.;Kim, Y.J.;Lee, H.T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.311-320
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    • 2002
  • The objective of this study was to assess the developmental potential in vitro produced embryos frozen-thawed with the various containers, and also examined expression and localization of heat shock protein 70 at these embryos. For the vitrification, 2-cell, 8-cell and blastocyst stage embryos produced by in vitro fertilization (IVF) and nuclear transfer (NT) were exposed the ethylene glycol 5.5 M freezing solution (EC 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop, and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min. and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid and cryo-loop. However, survival rates by straw were relatively lower than other containers. The use of cryo-loop resulted in only survival of nuclear transferred embryos (43.7%). Also, there embryos after IVF or NT were analysed by semi-quantitive reverse transcription-polymerase chain reaction (RT- PCR) methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNh were higher thawed embryos than control embryos. Immunocytochemistry used to localize the hsp 70 protein in embryos. Two and 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some frozen-thawed embryos. However, in the control, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform In distribution. Therefore, this result suggests that the exploiting Hsp 70 in the early embryos may be role for protection of stress condition for increase viability of embryos within IVF, NT and there frozen-thawed embryos.

돼지의 PSS 및 HSP70 유전자가 번식능력에 미치는 영향

  • 진현주;김인철;이장희;연성흠;김종대;조창연;정경용;정종현;위미순
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.59-59
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    • 2002
  • 돼지에서 스트레스관련 PSS(porcine stress syndrome) 및 HSP70(heat shock protein 70) 유전자형이 번식능력과 관련된 유전특성을 구명하기 위하여 PCR 용 프라이머의 염기서열을 설계하였다. PSS 는 Brenig등 (1992)의 RYR1 유전자 exon17 영역의 DNA에 근거한 18586-18659 영역과 GenBank (accession No. X68213) 의 HSP70 cDNA에 근거하여 290-512, 830-1424 및 1363-2041 영역으로 이루어졌다. (중략)

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Heat-Shock Protein 70 as a Tumor Antigen for in vitro Dendritic Cell Pulsing in Renal Cell Carcinoma Cases

  • Meng, Fan-Dong;Sui, Cheng-Guang;Tian, Xin;Li, Yan;Yang, Chun-Ming;Ma, Ping;Liu, Yun-Peng;Jiang, You-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8947-8950
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    • 2014
  • Immunological functions of heat shock proteins (HSPs) have long been recognized. In this study we aimed to efficiently purify HSP70 from renal cell carcinoma and test it as a tumor antigen for pulsing dendritic cells in vitro. HSP70 was purified from renal cell carcinoma specimens by serial column chromatography on Con A-sepharose, PD-10, ADP-agarose and DEAE-cellulose, and finally subjected to fast protein liquid chromatography (FPLC). Dendritic cells derived from the adherent fraction of peripheral blood mononuclear cells were cultured in the presence of IL-4 and GM-CSF and exposed to tumor HSP70. After 24 hours, dendritic cells were phenotypically characterized by flow cytometry. T cells obtained from the non-adherent fraction of peripheral blood mononuclear cells were then co-cultured with HSP70-pulsed dendritic cells and after 3 days T cell cytotoxicity towards primary cultured renal cell carcinoma cells was examined by Cell Counting Kit-8 assay. Dendritic cells pulsed in vitro with tumor-derived HSP70 expressed higher levels of CD83, CD80, CD86 and HLA-DR maturation markers than those pulsed with tumor cell lysate and comparable to that of dendritic cells pulsed with tumor cell lysate plus TNF-${\alpha}$. Concomitantly, cytotoxic T-lymphocytes induced by HSP70-pulsed dendritic cells presented the highest cytotoxic activity. There were no significant differences when using homologous or autologous HSP70 as the tumor antigen. HSP70 can be efficiently purified by chromatography and induces in vitro dendritic cell maturation in the absence of TNF-${\alpha}$. Conspecific HSP70 may effectively be used as a tumor antigen to pulse dendritic cells in vitro.

Evaluation of Thermo Tolerance of 'Nistari' an Indigenous Strain of Multivoltine Silkworm, Bombyx mori L.

  • Moorthy, S.M.;Das, S.K.;Mukhopadhyay, S.K.;Mandal, K.;Urs, S. Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.1
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    • pp.17-21
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    • 2007
  • An indigenous multivoltine silkworm, Nistari was evaluated for their thermo tolerance by exposing the larvae to various temperature regimes for eight hours. Among different temperature exposed, this strain has significant tolerance at $32^{\circ}C$. Analysis of heat shock protein revealed the expression of 70 kDa and 64 kDa polypeptides in fat body and midgut tissues. Interestingly esterase isozyme pattern in midgut showed characteristic expression of Est-1 and Est-3 at different temperatures signifying role in heat and cold shock.

The Expression of Heat Shock Protein in the Experimental Tooth Movement in Rats (백서의 실험적 치아이동시 열충격 단백의 발현)

  • Yoo, Dong-Whan;Kim, Eun-Cheol;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.31 no.2 s.85
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    • pp.249-259
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    • 2001
  • This study was designed to evaluate the expression of heat shock protein in tooth and surrounding tissue during the experimental movement of rat incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for heat shock protein. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats), to which 75g of force was applied from helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 0.5, 1, 4, 7, 14 and 28 days after force application, respectively. And the periodontal tissues of a control group and experimental groups were studied immunohistochemically. The results were as follows : 1. In control group, the expression of HSP47 was rare in gingiva, dentin and cementum, and mild in periodontal ligament and alveolar bone. But it was more evident than that of HSP70. 2. The expression of HSP47 or HSP70 was rare or mild in dentin, cementum and odontoblast of experimental group, regardless of the duration of force application, which was not different from that of control group. 3. In experimental group, the expression of HSP47 got to the highest degree in periodontal ligament and alveolar bone at 4 days after force application, and then decreased. And the expression was more evident in the pressure side than in the tension side of periodontal ligament 4. The expression of HSP70 began to increase at 12 hours after force application and got to the highest degree at 4 days, in the capillary of pulp and periodontal ligament. And the expression was more evident in the pressure side than in the tension side of periodontal ligament 5. The expression of HSP70 in alveolar bone of experimental group was rare, which was similar to that of control group.

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Sevoflurane Postconditioning Reduces Hypoxia/Reoxygenation Injury in Cardiomyocytes via Upregulation of Heat Shock Protein 70

  • Zhang, Jun;Wang, Haiyan;Sun, Xizhi
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1069-1078
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    • 2021
  • Sevoflurane postconditioning (SPostC) has been proved effective in cardioprotection against myocardial ischemia/reperfusion injury. It was also reported that heat shock protein 70 (HSP70) could be induced by sevoflurane, which played a crucial role in hypoxic/reoxygenation (HR) injury of cardiomyocytes. However, the mechanism by which sevoflurane protects cardiomyocytes via HSP70 is still not understood. Here, we aimed to investigate the related mechanisms of SPostC inducing HSP70 expression to reduce the HR injury of cardiomyocytes. After the HR cardiomyocytes model was established, the cells transfected with siRNA for HSP70 (siHSP70) or not were treated with sevoflurane during reoxygenation. The lactate dehydrogenase (LDH) level was detected by colorimetry while cell viability and apoptosis were detected by MTT and flow cytometry. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to detect HSP70, apoptosis-, cell cycle-associated factors, iNOS, and Cox-2 expressions. Enzyme-linked immuno sorbent assay (ELISA) was used to measure malondialdehyde (MDA) and superoxide dismutase (SOD). SPostC decreased apoptosis, cell injury, oxidative stress and inflammation and increased viability of HR-induced cardiomyocytes. In addition, SPostC downregulated Bax and cleaved caspase-3 levels, while SPostC upregulated Bcl-2, CDK-4, Cyclin D1, and HSP70 levels. SiHSP70 had the opposite effect that SPostC had on HR-induced cardiomyocytes. Moreover, siHSP70 further reversed the effect of SPostC on apoptosis, cell injury, oxidative stress, inflammation, viability and the expressions of HSP70, apoptosis-, and cell cycle-associated factors in HR-induced cardiomyocytes. In conclusion, this study demonstrates that SPostC can reduce the HR injury of cardiomyocytes by inducing HSP70 expression.

Difference of Gene Expression between Hypertrophic Scar Keratinocytes and Normal Keratinocytes (비후성 반흔 각질세포와 정상 각질세포의 유전자 비교분석)

  • Choi, Sung-Won;Chung, Ho-Yun;Lim, Young-Kook;Kim, Hoon-Nam;Oh, Ji-Won;Kim, Moon-Kyu;Jeon, Sae-Hwa;Hong, Yong-Taek
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.317-322
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    • 2010
  • Purpose: There is no clear evidence of the original cause of hypertrophic scar, and the effective method of treatment is not yet established. Recently the steps of searching in gene and molecular level are proceeding. we are trying to recognize the difference between keratinocytes of hypertrophic scar and normal skin. Then we do support the comprehension of the scar formation mechanism and scar management. Methods: Total RNAs were extracted from cultured keratinocytes from 4 hypertrophic scars and normal skins. The cDNA chips were prepared. A total of 3063 cDNAs from human cDNA library were arrayed. And the scanning data were analyzed. Results: On microarray, heat shock protein, pyruvate kinase, tumor rejection antigen were more than 2 fold intensity genes. Among them, heat shock 70 kd protein showed the strongest intensity difference. Conclusion: In this study, it can be concluded that heat shock proteins play an important role in the process of wound healing and scar formation. This study provides basic biologic information for scar research. The new way of the prevention and treatment of scar formation would be introduced with further investigations.

Analysis of heat, cold or salinity stress-inducible genes in the Pacific abalone, Haliotis discus hannai, by suppression subtractive hybridization

  • Nam, Bo-Hye;Park, Eun-Mi;Kim, Young-Ok;Kim, Dong-Gyun;Jee, Young-Ju;Lee, Sang-Jun;An, Cheul Min
    • The Korean Journal of Malacology
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    • v.29 no.3
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    • pp.181-187
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    • 2013
  • In order to investigate environmental stress inducible genes in abalone, we analyzed differentially expressed transcripts from a Pacific abalone, Haliotis discus hannai, after exposure to heat-, cold- or hyposalinity-shock by suppression subtractive hybridization (SSH) method. 1,074 unique sequences from SSH libraries were composed to 115 clusters and 986 singletons, the overall redundancy of the library was 16.3%. From the BLAST search, of the 1,316 ESTs, 998 ESTs (75.8%) were identified as known genes, but 318 clones (24.2%) did not match to any previously described genes. From the comparison results of ESTs pattern of three SSH cDNA libraries, the most abundant EST was different in each SSH library: small heat shock protein p26 (sHSP26) in heat-shock, trypsinogen 2 in cold-shock, and actin in hyposalinity SSH cDNA library. Based on sequence similarities, several response-to-stress genes such as heat shock proteins (HSPs) were identified commonly from the abalone SSH libraries. HSP70 gene was induced by environmental stress regardless of temperature-shock or salinity-stress, while the increase of sHSP26 mRNA expression was not detected in cold-shock but in heat-shock condition. These results suggest that the suppression subtractive hybridization method is an efficient way to isolate differentially expressed gene from the invertebrate environmental stress-response transcriptome.

Constitutive Expression of Small Heat Shock Protein Increases Thermotolerance in Transgenic Plant (저 분자량 Heat Shock Protein의 항상적 발현에 의한 형질전판 식물체의 고온내성 증가)

  • 이병현
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.13-18
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    • 2000
  • To investigate the function of chloroplast small HSP, transgenic tobacco plants (Nicotiana tabacum L. cv. Samsun) that constitutively overexpress the chloroplast small HSP (NtHSP21) from N. tabacum cv. Petit Havana SR1 were generated. Five homozygous lines of transformants showing different constitutive expression levels of the NtHSP21 were selected. To determine whether constitutive overexpression of NtHSP21 protein affects thermotolerance, wild-type and transformants were grown in Petri dishes, heat-stressed at 52$^{\circ}C$ for 45 min, and then incubated in normal growth condition. When heat-stressed wild-type plantlets were incubated at $25^{\circ}C$, leaf color gradually became white and all trio plantlets finally died within a week. As for the transformants, however, more than 70% of them remained green and survived under the conditions in which all the wild-type plants were dying. It was also found that the levels of NtHSP21 were correlated with the degree of thermotolerance. These results suggest that the NtHSP21 protein in transformants is responsible for the increase in thermotolerance.

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