Plasma exosomes are enriched in Hsp70 and modulated by stress and cortisol in rainbow trout

Exosomes are endosomally derived vesicles that are secreted from cells and contain a suite of molecules, including proteins and nucleic acids. Recent studies suggest the possibility that exosomes in circulation may be affecting recipient target cell function, but the modes of action are unclear. Here, we tested the hypothesis that exosomes are in circulation in fish plasma and that these vesicles are enriched with heat shock protein 70 (Hsp70). Exosomes were isolated from rainbow trout (Oncorhynchus mykiss) plasma using differential centrifugation, and their presence was confirmed by transmission electron microscopy and the exosomal marker acetylcholinesterase. Plasma exosomes were enriched with Hsp70, and this stress protein was transiently elevated in trout plasma in response to a heat shock in vivo. Using trout hepatocytes in primary culture, we tested whether stress levels of cortisol, the principle corticosteroid in teleosts, regulates exosomal Hsp70 content. As expected, a 1-h heat shock (+15°C above ambient) increased Hsp70 expression in hepatocytes, and this led to higher Hsp70 enrichment in exosomes over a 24-h period. However, cortisol treatment significantly reduced the expression of Hsp70 in exosomes released from either unstressed or heat-shocked hepatocytes. This cortisol-mediated suppression was not specific to Hsp70 as beta-actin expression was also reduced in exosomes released from hepatocytes treated with the steroid. Our results suggest that circulating Hsp70 is released from target tissues via exosomes, and their release is modulated by stress and cortisol. Overall, we propose a novel role for extracellular vesicular transport of Hsp70 in the organismal stress response.

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Glucocorticoid-mediated attenuation of the hsp70 response in trout hepatocytes involves the proteasome

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00125.2002 ◽

2002 ◽

Vol 283 (3) ◽

pp. R680-R687 ◽

Cited By ~ 36

Author(s):

Adrienne N. Boone ◽

Mathilakath M. Vijayan

Keyword(s):

Rainbow Trout ◽

Heat Shock ◽

Heat Shock Protein 70 ◽

Recovery Period ◽

Primary Cultures ◽

Hsp70 Expression ◽

Protein Breakdown ◽

Significant Drop ◽

Rainbow Trout Oncorhynchus Mykiss ◽

Hsp70 Synthesis

The physiological implication of elevated cortisol levels on cellular heat-shock protein 70 (hsp70) response was examined using primary cultures of rainbow trout ( Oncorhynchus mykiss) hepatocytes. Trout hepatocytes treated with cortisol, the predominant glucocorticoid in teleosts, responded to the heat shock (+15°C for 1 h) with a significant drop in hsp70 accumulation over a 24-h recovery period. [35S]methionine incorporation and pulse-chase studies confirmed that this cortisol impact was due to decreased hsp70 synthesis and not enhanced protein breakdown. Cortisol also significantly decreased glucocorticoid receptor (GR) expression in trout hepatocytes. This receptor downregulation was inhibited by the proteasomal inhibitors, lactacystin and MG-132, implying a role for the proteasome in GR downregulation by cortisol. Inhibiting the proteasome did not significantly modify heat-induced hsp70 accumulation in the absence of cortisol but significantly elevated hsp70 expression in the presence of cortisol in heat-shocked trout hepatocytes. Taken together, our results suggest proteasome-mediated GR degradation as a mechanism for the attenuation of hsp70 response by cortisol in heat-shocked hepatocytes.

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Effects ofHelicobacter pyloriand Heat Shock Protein 70 on the Proliferation of Human Gastric Epithelial Cells

Gastroenterology Research and Practice ◽

10.1155/2014/794342 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5 ◽

Cited By ~ 2

Author(s):

Liping Tao ◽

Hai Zou ◽

Zhimin Huang

Keyword(s):

Helicobacter Pylori ◽

Heat Shock ◽

Epithelial Cells ◽

Heat Shock Protein ◽

Mtt Assay ◽

Heat Shock Protein 70 ◽

Hsp70 Expression ◽

Gastric Epithelial Cells ◽

Ags Cells ◽

H Pylori

Infection ofHelicobacter pylori (H. pylori)changed the proliferation of gastric epithelial cells and decreased the expression of heat shock protein 70 (HSP70). However, the effects ofH. pylorion the proliferation of gastric epithelial cells and the roles of HSP70 during the progress need further investigation.Objective.To investigate the effects ofHelicobacter pylori (H. pylori)and heat shock protein 70 (HSP70) on the proliferation of human gastric epithelial cells.Methods. H. pyloriand a human gastric epithelial cell line (AGS) were cocultured. The proliferation of AGS cells was quantitated by an MTT assay, and the expression of HSP70 in AGS cells was detected by Western blotting. HSP70 expression in AGS cells was silenced by small interfering RNA (siRNA) to investigate the role of HSP70. ThesiRNA-treated AGS cells were cocultured withH. pyloriand cell proliferation was measured by an MTT assay.Results.The proliferation of AGS cells was accelerated by coculturing withH. pylorifor 4 and 8 h, but was suppressed at 24 and 48 h. HSP70 expression was decreased in AGS cells infected byH. pylorifor 48 h. The proliferation in HSP70-silenced AGS cells was inhibited after coculturing withH. pylorifor 24 and 48 h compared with the control group.Conclusions.Coculture ofH. pylorialtered the proliferation of gastric epithelial cells and decreased HSP70 expression. HSP70 knockdown supplemented the inhibitory effect ofH. pylorion proliferation of epithelial cells. These results indicate that the effects ofH. pylorion the proliferation of gastric epithelial cells at least partially depend on the decreased expression of HSP70 induced by the bacterium.

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Effects of ferulic acid on oxidative stress, heat shock protein 70, connexin 43, and monoamines in the hippocampus of pentylenetetrazole-kindled rats

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2016-0219 ◽

2017 ◽

Vol 95 (6) ◽

pp. 732-742 ◽

Cited By ~ 13

Author(s):

Abdelaziz M. Hussein ◽

Khaled M. Abbas ◽

Osama A. Abulseoud ◽

El-Hussainy M.A. El-Hussainy

Keyword(s):

Oxidative Stress ◽

Heat Shock ◽

Heat Shock Protein ◽

Ferulic Acid ◽

Group Treatment ◽

Heat Shock Protein 70 ◽

Connexin 43 ◽

Hsp70 Expression ◽

Neuroprotective Effects ◽

Cx43 Expression

The present study investigated the effects of ferulic acid (FA) on pentylenetetrazole (PTZ)-induced seizures, oxidative stress markers (malondialdehyde (MDA), catalase, and reduced glutathione (GSH)), connexin (Cx) 43, heat shock protein 70 (Hsp 70), and monoamines (serotonin (5-HT) and norepinephrine (NE)) levels in a rat model of PTZ-induced kindling. Sixty Sprague Dawley rats were divided into 5 equal groups: (a) normal group; (b) FA group: normal rats received FA at a dose of 40 mg/kg daily; (c) PTZ group: normal rats received PTZ at a dose of 50 mg/kg i.p. on alternate days for 15 days; (d) FA-before group: treatment was the same as for the PTZ group, except rats received FA; and (e) FA-after group: rats received FA from sixth dose of PTZ. PTZ caused a significant increase in MDA, Cx43, and Hsp70 along with a significant decrease in GSH, 5-HT, and NE levels and CAT activity in the hippocampus (p < 0.05). Pre- and post-treatment with FA caused significant improvement in behavioral parameters, MDA, CAT, GSH, 5-HT, NE, Cx43 expression, and Hsp70 expression in the hippocampal region (p < 0.05). We conclude that FA has neuroprotective effects in PTZ-induced epilepsy, which might be due to attenuation of oxidative stress and Cx43 expression and upregulation of neuroprotective Hsp70 and neurotransmitters (5-HT and NE).

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Επικεκαλυμμένες ενδαγγειακές προθέσεις και επαναστένωση μετά από αγγειοπλαστική

10.12681/eadd/36988 ◽

2014 ◽

Author(s):

Δημήτριος Λυσίτσας

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Heat Shock Protein 70 ◽

Low Dose ◽

High Dose ◽

Hsp 70

Εισαγωγή: Η υπερπλασία του έσω χιτώνα παίζει μείζων ρόλο στην επαναστένωση (in-stentrestenosis). Στην παρούσα μελέτη αξιολογήσαμε in vitro την επίδραση της D-24851(κυτταροτοξική ουσία που σταματά τον κυτταρικό κύκλο στο στάδιο G2-M) στονπολλαπλασιασμό των λείων μυϊκών κυττάρων και μελετήσαμε την ασφάλεια και τηνδραστικότητα μίας ενδαγγειακής πρόθεσης (stent) επικαλυμμένης με πολυμερή ουσία πουαπελευθερώνει την D-24851, στην αναστολή της υπερπλασίας του έσω χιτώνα χωρίς ναεμποδίζει την αναγεννητική ικανότητα του ενδοθηλίου σε in vivo πειραματικό μοντέλο.Υλικό και Μέθοδοι: Γυμνά μεταλλικά stent (n=6), stent επικαλυμμένα μόνο με πολυμερήουσία (polymer-coated, n=7) και stent επικαλυμμένα με πολυμερή ουσία πουαπελευθερώνουν 31±1μg (low-dose, n=7), 216±8 μg (high-dose, n=6) ή 1774±39 μg(extreme-dose, n=5) της D-24851 εμφυτεύτηκαν στις μηριαίες αρτηρίες λευκών New Zealandκουνελιών. Τα πειραματόζωα θυσιάστηκαν στις 28 ημέρες για ιστομορφομετρική ανάλυση.Για την αξιολόγηση της ενδοθηλιακής αναγέννησης στις 90 ημέρες, 12 πειραματόζωαχρησιμοποιήθηκαν για την τοποθέτηση polymer-coated (n=3), low dose (n=3), high dose(n=3) or extreme dose (n=3) ενδαγγειακών προθέσεων.Αποτελέσματα: In vitro η D-24851 αναστέλλει την υπερπλασία των λείων μυϊκών κυττάρωνκαι επάγει την απόπτωση τους χωρίς να αυξάνει την επαγωγή της heat shock protein 70(HSP-70), μία κυτταροπροστατευτική και αντι-αποπτωτική πρωτεΐνη. Η θεραπεία με lowdoseD-24851 stents συνδυάστηκε με 38% (P=0.029) μείωση της υπερπλαστικής περιοχήςτου έσω χιτώνα και 35% (P=0.003) μείωση της επι τοις εκατό στένωσης του αυλού σεσύγκριση με τα γυμνά μεταλλικά stents. Ο τραυματισμός και η φλεγμονή του αρτηριακού τοιχώματος δεν παρουσίασαν σημαντικές διαφορές μεταξύ των ομάδων. Τα επικαλυμμέναμόνο με πολυμερή ουσία stents εμφάνισαν παρόμοια ανάπτυξη νεοιστού σε σύγκριση με ταγυμνά μεταλλικά stents. Ωστόσο, όλες οι ομάδες των stents με D-24851 παρουσίασαν ατελήενδοθηλιοποίηση συγκρινόμενα με τα polymer-coated stents.Συμπεράσματα: Οι επικεκαλυμμένες ενδαγγειακές προσθέσεις με πολυμερή ουσία καιχαμηλη δόση D-24851 μειώνουν σημαντικά την υπερπλασιά του έσω χιτώνα. Λόγω τηςατελούς ενδοθηλιοποίησης, μακράς διάρκειας μελέτες είναι απαραίτητες για ναπιστοποιήσουν ότι η αναστολή του νεοιστού παραμένει και μετά τις 28 ημέρες.

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The effect of red-fleshed pitaya (Hylocereus polyrhizus) on heat shock protein 70 and cortisol expression in strenuous exercise induced rats

F1000Research ◽

10.12688/f1000research.17533.1 ◽

2019 ◽

Vol 8 ◽

pp. 130

Author(s):

Novita Sari Harahap ◽

Aznan Lelo ◽

Ambrosius Purba ◽

Awaluddin Sibuea ◽

Rina Amelia ◽

...

Keyword(s):

Body Weight ◽

Heat Shock ◽

Heat Shock Protein ◽

Heat Shock Protein 70 ◽

The Body ◽

Male Rats ◽

Strenuous Exercise ◽

Hsp70 Expression ◽

Average Weight ◽

Significant Difference

Background: Oxidative stress from exercise can contribute to damaging cells, increasing heat shock protein 70 (HSP70) and suppressing the immune system in the body. This research aimed to determine the antioxidant potential of red-fleshed pitaya extract on HSP70 and cortisol expression in rats which were subjected to strenuous exercise. Methods: The subjects of this research were 32 Sprague Dawley male rats, aged 3 months, with an average weight of 200 g. Red-fleshed pitaya extract was obtained from methanol extraction process; a maceration technique was performed and the extract was concentrated using an air-drying method. Rats were randomly divided into four groups. Group 1 were subjected to strenuous exercise and treated with distilled water only; while Groups 2, 3 and 4 were subjected to strenuous exercise and treated with 100 mg/kg body weight, 200 mg/kg body weight and 300 mg/kg body weight of red-fleshed pitaya extract, respectively. Strenuous exercises in rats was performed by intense swimming of 20 min/day, 3 days a week for 3 weeks. HSP70 expression and cortisol were measured with Enzyme-Linked Immune Sorbent Assay (ELISA) method. Results: There was a significant reduction of HSP70 (p=0.000) and cortisol expression (p=0.000) between the groups. Also, there was a significant difference in the average decreasing of HSP70 expression between group 4 and either groups 1 or 2 (p=0.000). However, a significant difference between groups 4 and 3 was not observed (p=0.813). Lastly, a significant difference was found in the average decrease of cortisol expression between groups 4 and 1 (p=0.000), 2 (p=0.000), and 3 (p=0.000) respectively. Conclusion: Red-fleshed pitaya is potential to be utilized as antioxidant to decrease the HSP70 and cortisol expression.

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Expression of heat shock protein 70 is insufficient to extend Drosophila melanogaster longevity

10.1101/753269 ◽

2019 ◽

Author(s):

Chengfeng Xiao ◽

Danna Hull ◽

Shuang Qiu ◽

Joanna Yeung ◽

Jie Zheng ◽

...

Keyword(s):

Heat Treatment ◽

Drosophila Melanogaster ◽

Heat Shock ◽

Heat Shock Protein ◽

Stress Resistance ◽

Heat Shock Protein 70 ◽

Biological Effect ◽

Hsp70 Expression ◽

Gene Encoding ◽

Shock Proteins

AbstractIt has been known for over 20 years that Drosophila melanogaster flies with twelve additional copies of the hsp70 gene encoding the 70 kDa heat shock protein lives longer after a non-lethal heat treatment. Since the heat treatment also induces the expression of additional heat shock proteins, the biological effect can be due either to HSP70 acting alone or in combination. This study used the UAS/GAL4 system to determine whether hsp70 is sufficient to affect the longevity and the resistance to thermal, oxidative or desiccation stresses of the whole organism. We observed that HSP70 expression in the nervous system or muscles has no effect on longevity or stress resistance but ubiquitous expression reduces the life span of males. We also observed that the down-regulation of Hsp70 using RNAi did not affect longevity.

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Synthesis of stress protein 70 (Hsp70) in rainbow trout (Oncorhynchus mykiss) red blood cells

Journal of Experimental Biology ◽

10.1242/jeb.200.3.607 ◽

1997 ◽

Vol 200 (3) ◽

pp. 607-614 ◽

Cited By ~ 6

Author(s):

S Currie ◽

B Tufts

Keyword(s):

Protein Synthesis ◽

Rainbow Trout ◽

Heat Shock ◽

Oncorhynchus Mykiss ◽

Red Blood Cells ◽

Blood Cells ◽

Stress Proteins ◽

Stress Protein ◽

Rainbow Trout Oncorhynchus Mykiss ◽

Hsp70 Synthesis

Unlike enucleated mammalian red blood cells (rbcs), the nucleated rbcs of lower vertebrates are capable of protein synthesis and may, therefore, serve as a valuable model to investigate the adaptive significance of stress protein synthesis in cells. This study examined the synthesis of stress protein 70 (Hsp70) in rbcs of the temperature-sensitive rainbow trout Oncorhynchus mykiss in response to heat shock and anoxia. Through western blot analysis, we have demonstrated that rainbow trout rbcs synthesize Hsp70 both constitutively and in response to an increase in temperature. Radioisotopic labelling experiments indicated that the temperature at which Hsp70 synthesis was induced in fish acclimated to 10 °C was between 20 and 25 °C. Actinomycin D blocked de novo Hsp70 synthesis, implying that synthesis of Hsp70 is regulated at the level of transcription in rainbow trout rbcs. Since trout rbcs rely heavily on aerobic metabolism, but may also experience very low oxygen levels within the circulation, we also examined the relative importance of (1) anoxia as a stimulus for Hsp70 synthesis and (2) oxygen as a requirement for protein synthesis under control and heat-shock conditions. We found that trout rbcs were capable of protein synthesis during 2 h of anoxia, but did not increase Hsp70 synthesis. Moreover, rbcs subjected to combined anoxia and heat shock exhibited increases in Hsp70 synthesis that were similar in magnitude to those in cells exposed to heat shock alone. The latter results suggest that rainbow trout rbcs are (1) able to synthesize non-stress proteins during anoxia, (2) capable of tolerating periods of reduced oxygen availability without increased synthesis of stress proteins and (3) able to maintain the integrity of their heat-shock response even during periods of anoxia.

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Heat shock protein 70 / MAGE-1 tumor vaccine can enhance the potency of MAGE-1–specific cellular immune responses in vivo

Cancer Immunology Immunotherapy ◽

10.1007/s00262-004-0536-6 ◽

2004 ◽

Vol 53 (9) ◽

pp. 825-834 ◽

Cited By ~ 13

Author(s):

Jing Ye ◽

Guang-Sheng Chen ◽

Hong-Ping Song ◽

Zeng-Shan Li ◽

Ya-Yu Huang ◽

...

Keyword(s):

Heat Shock ◽

Immune Responses ◽

Heat Shock Protein ◽

Heat Shock Protein 70 ◽

Tumor Vaccine ◽

Cellular Immune Responses ◽

Cellular Immune

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In Vivo Chaperone Activity of Heat Shock Protein 70 and Thermotolerance

Molecular and Cellular Biology ◽

10.1128/mcb.19.3.2069 ◽

1999 ◽

Vol 19 (3) ◽

pp. 2069-2079 ◽

Cited By ~ 145

Author(s):

Ellen A. A. Nollen ◽

Jeanette F. Brunsting ◽

Han Roelofsen ◽

Lee A. Weber ◽

Harm H. Kampinga

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Heat Resistance ◽

Cell Line ◽

Heat Shock Protein 70 ◽

Chaperone Activity ◽

Luciferase Reporter ◽

Reporter Protein ◽

Regulated Gene Expression

ABSTRACT Heat shock protein 70 (Hsp70) is thought to play a critical role in the thermotolerance of mammalian cells, presumably due to its chaperone activity. We examined the chaperone activity and cellular heat resistance of a clonal cell line in which overexpression of Hsp70 was transiently induced by means of the tetracycline-regulated gene expression system. This single-cell-line approach circumvents problems associated with clonal variation and indirect effects resulting from constitutive overexpression of Hsp70. The in vivo chaperone function of Hsp70 was quantitatively investigated by using firefly luciferase as a reporter protein. Chaperone activity was found to strictly correlate to the level of Hsp70 expression. In addition, we observed an Hsp70 concentration dependent increase in the cellular heat resistance. In order to study the contribution of the Hsp70 chaperone activity, heat resistance of cells that expressed tetracycline-regulated Hsp70 was compared to thermotolerant cells expressing the same level of Hsp70 plus all of the other heat shock proteins. Overexpression of Hsp70 alone was sufficient to induce a similar recovery of cytoplasmic luciferase activity, as does expression of all Hsps in thermotolerant cells. However, when the luciferase reporter protein was directed to the nucleus, expression of Hsp70 alone was not sufficient to yield the level of recovery observed in thermotolerant cells. In addition, cells expressing the same level of Hsp70 found in heat-induced thermotolerant cells containing additional Hsps showed increased resistance to thermal killing but were more sensitive than thermotolerant cells. These results suggest that the inducible form of Hsp70 contributes to the stress-tolerant state by increasing the chaperone activity in the cytoplasm. However, its expression alone is apparently insufficient for protection of other subcellular compartments to yield clonal heat resistance to the level observed in thermotolerant cells.

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