• Title/Summary/Keyword: Heat-shock protein

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Vascular Smooth Muscle Cells Secrete CXCL10 in Response to Heat Shock Protein 90 (혈관평활근세포에서 HSP90에 의한 CXCL10 증가에 관여하는 인자에 대한 연구)

  • Rhim, Byung-Yong;Kim, Do-Hyung;Kim, Koan-Hoi
    • Journal of Life Science
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    • v.21 no.5
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    • pp.664-670
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    • 2011
  • Oxidative stress results in sustained release of heat shock protein 90 (HSP90) from vascular smooth muscle cells (VSMCs). We investigated whether extracellular HSP90 predisposed VSMCs to pro-inflammatory phenotype. Exposure of human aortic smooth muscle cells to HSP90 not only significantly enhanced CXCL10 secretion but also increased CXCL10 transcription. HSP90-mediated CXCL10 secretion was attenuated by OxPAPC, a TLR-2/4 inhibitor, and curcumin, a TLR-4 dimerization inhibitor. Inhibitors of diphenyleneiodium chloride and the Akt pathway also attenuated CXCL10 secretion in response to HSP90. The gene delivery of I${\kappa}$B using recombinant adenoviruses and treatment with resveratrol, which inhibit NF-${\kappa}$B activity, significantly attenuated HSP90-induced CXCL10 secretion from VSMCs. We propose that extracellular HSP90 contributes to an inflammatory reaction in the stressed vasculature by inducing CXCL10 expression of VSMCs, and that TLR-4, Akt, and NF-${\kappa}$B play active roles in the process.

Heat shock protein 90 inhibitor AUY922 attenuates platelet-derived growth factor-BB-induced migration and proliferation of vascular smooth muscle cells

  • Kim, Jisu;Lee, Kang Pa;Kim, Bom Sahn;Lee, Sang Ju;Moon, Byung Seok;Baek, Suji
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.3
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    • pp.241-248
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    • 2020
  • Luminespib (AUY922), a heat shock proteins 90 inhibitor, has anti-neoplastic and antitumor effects. However, it is not clear whether AUY922 affects events in vascular diseases. We investigated the effects of AUY922 on the platelet-derived growth factor (PDGF)-BB-stimulated proliferation and migration of vascular smooth muscle cells (VSMC). VSMC viability was detected using the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) reagent. To detect the attenuating effects of AUY922 on PDGF-BB-induced VSMCs migration in vitro, we performed the Boyden chamber and scratch wound healing assays. To identify AUY922-mediated changes in the signaling pathway, the phosphorylation of protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) 1/2 was analyzed by immunoblotting. The inhibitory effects of AUY922 on migration and proliferation ex vivo were tested using an aortic ring assay. AUY922 was not cytotoxic at concentrations up to 5 nM. PDGF-BB-induced VSMC proliferation, migration, and sprout outgrowth were significantly decreased by AUY922 in a dose-dependent manner. AUY922 significantly reduced the PDGF-BB-stimulated phosphorylation of Akt and ERK1/2. Furthermore, PD98059 (a selective ERK1/2 inhibitor) and LY294002 (a selective Akt inhibitor) decreased VSMC migration and proliferation by inhibiting phosphorylation of Akt and ERK1/2. Greater attenuation of PDGF-BB-induced cell viability and migration was observed upon treatment with PD98059 or LY294002 in combination with AUY922. AUY922 showed anti-proliferation and anti-migration effects towards PDGF-BB-induced VSMCs by regulating the phosphorylation of ERK1/2 and Akt. Thus, AUY922 is a candidate for the treatment of atherosclerosis and restenosis.

High Temperature Stress of Summer Chinese Cabbage in Alpine Region (고랭지 여름배추의 고온장해 원인 해석)

  • Hwang, Seon-Woong;Lee, Ju-Young;Hong, Sung-Chang;Park, Yang-Ho;Yun, Seung-Gil;Park, Moon-Hee
    • Korean Journal of Soil Science and Fertilizer
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    • v.36 no.6
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    • pp.417-422
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    • 2003
  • The objective of this study was to assess the regime of heat shock protein and leaf temperature caused by high temperature stress in chinese cabbage grown in alpine region. In monsoon period, high temperature and heavy rain have caused a stress condition for the cultured higher plants. Chinese cabbages were grown in different altitude, i.e. 600 m and 1,100 m. It was demonstrated that heat shock protein (Hsp 90) in alpine chinese cabbage leaf was actively expressed by high temperature and surplus nitrogen application. As a results of thermo-graphically observed leaf temperatures, chinese cabbage grown in high altitude region were ranged from 20.5 to $24.3^{\circ}C$ while in low altitude from 24.0 to $31.5^{\circ}C$. Furthermore, analysis of assimilated nutrients indicated that total nitrogen content was higher in plant grown under high temperature than under low temperature.

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.

Expression of the Heat Shock Protein 70 Gene and External Developmental Traits of Two Bivalvia Species, Crassostrea gigas and Mytilus galloprovincialis, under Aquaculture Environments (사육환경에 따른 이매패류 (Crassostrea gigas, Mytilus galloprovincialis)의 외부형질 성장과 Heat Shock Protein 70 유전자 발현)

  • Kim, Won-Seok;Park, Kiyun;Kim, Jong Kyu;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.49 no.1
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    • pp.22-30
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    • 2016
  • Environmental changes exert harmful effects on organisms inhabiting coastal regions. These changes are also associated with reduced production in aquaculture farms. In this study, we investigated internal and external responses of two Bivalvia species (Crassostrea gigas and Mytilus galloprovincialis) in Gamak Bay under stressful environmental conditions in aquaculture farms. We investigated external responses such as weight, size, and environment exposure time, and analyzed the expression of the HSP70 gene. C. gigas HSP70 gene expression level was significantly high in the C3 aquaculture farm site, but the weight and size of C. gigas were high in the C2 aquaculture farm site. The response of C. gigas HSP70 mRNA was associated with the environmental exposure time in each aquaculture farm. Expression of M. galloprovincialis HSP70 gene was found to be significantly higher in the M2 aquaculture farm site than in the M1 site, whereas the weight of M. galloprovincialis was observed to be higher in the M1 site. The size and environmental exposure time of M. galloprovincialis were similar between M1 and M2 sites. In addition, HSP70 sequences of C. gigas and M. galloprovincialis showed high similarity with that of another marine species. According to our results, there were differences in internal responses following environmental stress in aquaculture farms, with respect to HSP70 gene expression. The results suggest that the HSP70 gene is a useful molecular indicator for monitoring stress responses in Bivalvia species in the field.

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.

Anti-inflammatory effects of Grateloupia elliptica Fermenting Extracts Using Aureobasidium pullulans (흑효모를 이용한 참도박 발효 추출물의 항염 효과)

  • Vu, Van Vinh;Lee, Kyung Eun;Kang, Sang Gu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.2
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    • pp.123-131
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    • 2021
  • In this study, we investigated the biological functions of Grateloupia elliptica (G. elliptica) fermented with Aureobasidium pullulans (A. pullulans). Total phenolic contents (TPC) of the hot-water extract of the fermented G. elliptica increased 2.7 folds than that of the non-fermented G. elliptica. Furthermore, total flavonoid contents of both the hot-water extract and the ethanol extract increased maximum 2.4 folds amounts than non-fermented G. elliptica extracts.HaCaT cells were induced inflammation treated with LPS (1 ㎍/mL) or H2O2 (1mM) and examined with 100 ㎍/mL of G. elliptica extracts. The extraction of the fermented G. elliptica increased HaCaT cell proliferation in the maximum 10% than non-fermented G. elliptica extraction. Furthermore, investigating changes in protein expression associated with inflammation resulted in a significant reduction in the expression of cyclooxygenase-2 and 70 kDa heat shock proetin. Conclusively, the extracts of G. elliptica fermented with A. pullulans have bioactive functions both anti-oxidant to protect environmental stresses and anti-inflammation activity. Hence, G. elliptica fermented with A. pullulans would be a good natural resource as bioactive ingredients for cosmetics. Therefore, G. elliptica fermented with A. pullulans is useful as a astringent material with anti-inflammatory skin.

Heat Shock Protein 90 Gene Expression in Juvenile Sea Cucumber Apostichopus japonicus (Echinodermata; Holothuroidea) according to Releasing Methods (어린 돌기해삼 Apostichopus japonicus (Echinodermata; Holothuroidea) 방류 방법에 따른 열충격단백질90 유전자의 발현 분석)

  • Lee, Dong-Han;Lee, Seungheon;Jeong, Dong-Bin;Sohn, Young Chang
    • Journal of Marine Life Science
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    • v.7 no.1
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    • pp.29-36
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    • 2022
  • Sea cucumber, Aposticopus japonicus, is a major invertebrate species in the coastal regions of Korea. To evaluate the short-term stress levels according to the releasing methods, this study investigated the gene expression profiles of heat shock protein 90 (HSP90) by real-time quantitative polymerase chain reaction. When the juvenile sea cucumbers were packed in the vinyl bag with oxygen followed by transportation for 30 min or air-exposed for 1 h, the HSP90 gene expression levels in the experimental groups were significantly increased compared to those of the control groups (transported group, p=0.001; air-exposed group, p=0.032). The experimental group at 6 h post-release by seed-spreading method and at 2~6 h post-release by underwater hose-releasing method on board a fishing boat showed that the levels of HSP90 gene expression were not statistically significant but decreased slightly compared to the control group (seed-spreading group, p=0.069; hose-releasing group, p=0.093). On the other hand, the HSP90 gene expression showed an increasing pattern as the time passed (~6 h) after underwater release of juvenile sea cucumbers by divers (p=0.061). These results suggest that HSP90 gene expression can be used to investigate short-term stress response and effective releasing methods of juvenile sea cucumbers.

Heat Shock Protein Association with Clinico-Pathological Characteristics of Gastric Cancer in Jordan : HSP70 is Predictive of Poor Prognosis

  • Bodoor, Khaldon;Jalboush, Sara Abu;Matalka, Ismail;Abu-Sheikha, Aya;Waqfi, Rofieda Al;Ebwaini, Hanadi;Abu-Awad, Aymen;Fayyad, Luma;Al-Arjat, Jamal;Haddad, Yazan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3929-3937
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    • 2016
  • Gastric cancer (GC) is a major health problem worldwide and is one of the ten most commonly diagnosed cancers in Jordan. GC is usually diagnosed at late aggressive stages in which treatment options are limited. Recently, heat shock proteins (HSPs) were found to be overexpressed in a wide range of malignancies have been considered as promising candidate biomarkers for GC. The aim of this study was to investigate pathogenic roles of a panel of cytosolic HSPs including HSP90, HSP70, HSP60 and HSP27 in GC. Immunohistochemistry was used to assess the level of expression of these proteins in archived tumor samples (N=87) representing various pathological characteristics of GC. HSP90, HSP60 and HSP27 were expressed abundantly in gastric tumors. On the other hand, HSP70 was reduced significantly and also found to be associated with Helicobacter pylori infection in tissues collected from GC patients. Furthermore, HSP27 was found to be associated with the level of differentiation. Our findings indicate a role of HSP70 as a potential prognostic biomarker, patients harboring positive HSP70 expression displaying worse disease free survival than those with negative HSP70 expression. Differential expression of HSPs may play crucial roles in the initiation and progression of GC, and could be exploited as future therapeutic targets.

Proteomic Analysis of Protein Expression in Streptococcus pneumoniae in Response to Temperature Shift

  • Lee Myoung-Ro;Bae Song-Mee;Kim Tong-Soo;Lee Kwang-Jun
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.375-382
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    • 2006
  • From its initial colonization to causation of disease, Streptococcus pneumoniae has evolved strategies to cope with a number of stressful in vivo environmental conditions. In order to analyze a global view of this organism's response to heat shock, we established a 2-D electrophoresis proteome map of the S. pneumoniae D39 soluble proteins under in vitro culture conditions and performed the comparative proteome analysis to a 37 to $42^{\circ}C$ temperature up-shift in S. pneumoniae. When the temperature of an exponentially growing S. pneumoniae D39 culture was raised to $42^{\circ}C$, the expression level of 25 proteins showed changes when compared to the control. Among these 25 proteins, 12 were identified by MALDI-TOF and LC-coupled ESI MS/MS. The identified proteins were shown to be involved in the general stress response, energy metabolism, nucleotide biosynthesis pathways, and purine metabolism. These results provide clues for understanding the mechanism of adaptation to heat shock by S. pneumoniae and may facilitate the assessment of a possible role for these proteins in the physiology and pathogenesis of this pathogen.