Effect Of Heat Shock Protein Research Articles

Cytoprotective Effect of Heat Shock Protein 27 Against Lipopolysaccharide-Induced Apoptosis of Renal Epithelial HK-2 Cells

Background: In response to various stimuli, heat shock protein 27 (Hsp27) functions as an anti-apoptotic or/and anti-inflammatory factor which confers a survival advantage to cells. This study was aimed to explore whether Hsp27 also has a cytoprotective role in human renal tubular epithelial cells, and to evaluate its potential in treating septic acute kidney injury (septic AKI). Methods: HK-2 cells were subjected to different concentrations (0-10 µg/mL) of lipopolysaccharide (LPS) for various times (0-24 h) to establish a septic AKI model in vitro. Before LPS administration, HK-2 cells were transfected either with vectors or siRNA against Hsp27, and the changes in cell viability and apoptotic cells rate were assessed using CCK-8 and flow cytometry. The expression changes in apoptosis-related proteins, proinflammatory cytokines and chemokine, as well as main factors in NF-κB and JNK pathways were mainly determined by Western blotting. Besides, the relationship between Hsp27 and Bcl-2 was detected by co-immunoprecipitation. Results: LPS remarkably damaged HK-2 cells by reduction of cell viability, induction of apoptosis, and stimulation of proinflammatory cytokines and chemokine release. Hsp27 overexpression significantly impaired LPS-induced damage in HK-2 cells. Hsp27 overexpression couldn’t alter the mRNA level of Bcl-2, but could interact with Bcl-2 at an endogenous level. Both NF-κB and JNK pathways were activated by LPS, while were blocked in Hsp27-overexpressing cells. Conclusion: Hsp27 overexpression conferred a survival advantage to LPS-injured HK-2 cells by controlling cell viability, apoptosis and inflammation, possibly via interaction with Bcl-2 and modulation of NF-κB and JNK pathways.

Extracellular HSP70/HSP70-PCs regulate hepatocarcinoma cell migration and invasion via RhoA.

The effects of heat shock protein 70 (HSP70)/HSP70-peptide complexes (HSP70-PCs) on the invasion and metastasis ability of hepatocellular carcinoma (HCC) Huh-7 cells were investigated. Wound healing assay revealed that cells treated with HSP70/HSP70-PCs healed faster than negative control cells. HSP70/HSP70-PCs-treated cells also exhibited better migration ability and higher invasion ability than control cells. HSP70/HSP70-PCs treatment did not alter the messenger RNA (mRNA) or protein levels of matrix metalloproteinase-9; the opposite was true for Ras homolog family member A (RhoA) mRNA and protein levels. RNA interference of RhoA attenuated the migration of HSP70/HSP70-PCs-treated cells. The present findings indicate that regulation of HCC cell migration by HSP70/HSP70-PCs occurs via regulation of RhoA expression.

Reduced Contractility and Motility of Prostatic Cancer-Associated Fibroblasts after Inhibition of Heat Shock Protein 90

Background: Prostate cancer-associated fibroblasts (CAF) can stimulate malignant progression and invasion of prostatic tumour cells via several mechanisms including those active in extracellular matrix; Methods: We isolated CAF from prostate cancer patients of Gleason Score 6–10 and confirmed their cancer-promoting activity using an in vivo tumour reconstitution assay comprised of CAF and BPH1 cells. We tested the effects of heat shock protein 90 (HSP90) inhibitors upon reconstituted tumour growth in vivo. Additionally, CAF contractility was measured in a 3D collagen contraction assay and migration was measured by scratch assay; Results: HSP90 inhibitors dipalmitoyl-radicicol and 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG) reduced tumour size and proliferation in CAF/BPH1 reconstituted tumours in vivo. We observed that the most contractile CAF were derived from patients with lower Gleason Score and of younger age compared with the least contractile CAF. HSP90 inhibitors radicicol and 17-DMAG inhibited contractility and reduced the migration of CAF in scratch assays. Intracellular levels of HSP70 and HSP90 were upregulated upon treatment with HSP90 inhibitors. Inhibition of HSP90 also led to a specific increase in transforming growth factor beta 2 (TGFβ2) levels in CAF; Conclusions: We suggest that HSP90 inhibitors act not only upon tumour cells, but also on CAF in the tumour microenvironment.

Intracellular formation of α-synuclein oligomers and the effect of heat shock protein 70 characterized by confocal single particle spectroscopy

Synucleinopathies such as dementia with Lewy bodies or Parkinson’s disease are characterized by intracellular deposition of pathologically aggregated α-synuclein. The details of the molecular pathogenesis of PD and especially the conditions that lead to intracellular aggregation of α-synuclein and the role of these aggregates in cell death remain unknown. In cell free in vitro systems considerable knowledge about the aggregation processes has been gathered. In comparison, the knowledge about these aggregation processes in cells is far behind. In cells α-synuclein aggregates can be toxic. However, the crucial particle species responsible for decisive steps in pathogenesis such as seeding a continuing aggregation process and triggering cell death remain to be identified. In order to understand the complex nature of intracellular α-synuclein aggregate formation, we analyzed fluorescent particles formed by venus and α-synuclein-venus fusion proteins and α-synuclein-hemi-venus fusion proteins derived from gently lyzed cells. With these techniques we were able to identify and characterize α-synuclein oligomers formed in cells. Especially the use of α-synuclein-hemi-venus fusion proteins enabled us to identify very small α-synuclein oligomers with high sensitivity. Furthermore, we were able to study the molecular effect of heat shock protein 70, which is known to inhibit α-synuclein aggregation in cells. Heat shock protein 70 does not only influence the size of α-synuclein oligomers, but also their quantity. In summary, this approach based on fluorescence single particle spectroscopy, that is suited for high throughput measurements, can be used to detect and characterize intracellularly formed α-synuclein aggregates and characterize the effect of molecules that interfere with α-synuclein aggregate formation.

Detrimental effects of geldanamycin on adults and larvae ofTrichinella spiralis

SummaryTrichinellosis is a zoonotic disease affecting mainly the temperate regions. The treatment is a challenge for the physician, and the available therapy is far from ideal. Therefore, this work aimed to evaluate the effect of heat shock protein 90 inhibitor, geldanamycin, on the adult worms and larvae ofTrichinella spiralis. This research comprised anin vivostudy in whichT. spiralis-infected mice were treated by two different doses of geldanamycin, thereafter larval count and pathological changes were determined in the muscles. Meanwhile, thein vitrostudy investigated the effect of two different concentrations of geldanamycin on adult worms and larvae ofT. spiralisvia transmission electron microscopy. Thein vivostudy showed significant reduction of muscle larval counts under the effect of geldanamycin. Moreover, characteristic changes were noted as regards the parasite and the inflammatory response. Thein vitrostudy revealed degenerative changes in the body wall of larvae and adults ofT. spiralisunder the influence of geldanamycin. In conclusion, heat shock protein 90 inhibitor, geldanamycin, seems to have detrimental effects on the adults and larvae ofT. spiralis. It, or one of its derivatives, could be an adjuvant to anthelmintic therapy of trichinellosis, but more studies are warranted to establish its usefulness.

The Effect of Heat Shock Protein 70 (HSP70) on the Activity of Notch Signaling Pathway in Myelin-Reactive CD4 T Cells (P5.332)

The Effect of Heat Shock Protein 70 (HSP70) on the Activity of Notch Signaling Pathway in Myelin-Reactive CD4 T Cells (P5.332)

Effect of heat shock protein 70 polymorphism on thermotolerance in Tharparkar cattle.

Aim:Out of various members of heat shock protein (HSP) superfamily which act a molecular chaperon by binding to the denaturing protein thus stabilizing them and preserving their activity, HSP70 are of major importance in thermotolerance development. Thus, present investigation aimed at a screening of HSP70 gene for polymorphisms and possible differences in thermotolerance in Tharparkar breed of cattle.Materials and Methods:A 295 bp fragment of HSP70 gene was subjected to polymerase chain reaction-single-strand conformation polymorphism (SSCP) followed by sequencing of different SSCP patterns in 64 Tharparkar cattle. A comparative thermotolerance of identified genotypes was analyzed using heat tolerance coefficients (HTCs) of animals for different seasons.Results:Three SSCP patterns and consequently two alleles namely A and B were documented in one fragment of HSP70 gene. On sequencing, one single-nucleotide polymorphism with G > T substitution was found at a position that led to a change of amino acid aspartate to tyrosine in allele A. It was found that in maintaining near normal average rectal temperature, genotype AA was superior (p≤0.01). Genotype AA, thus, was found to be most thermotolerant genotype with the highest HTC (p≤0.01).Conclusion:The polymorphism at HSP70 is expected to be a potent determinant for heat tolerance in cattle, which may aid in selection for thermotolerance in cattle.

Beyond Thermal Performance Curves: Modeling Time-Dependent Effects of Thermal Stress on Ectotherm Growth Rates.

Thermal performance curves have been widely used to model the ecological responses of ectotherms to variable thermal environments and climate change. Such models ignore the effects of time dependence-the temporal pattern and duration of temperature exposure-on performance. We developed and solved a simple mathematical model for growth rate of ectotherms, combining thermal performance curves for ingestion rate with the temporal dynamics of gene expression and protein production in response to high temperatures to predict temporal patterns of growth rate in constant and diurnally fluctuating temperatures. We used the model to explore the effects of heat shock proteins on larval growth rates of Manduca sexta. The model correctly captures two empirical patterns for larval growth rate: first, maximal growth rate and optimal temperature decline with increasing duration of temperature exposure; second, mean growth rates decline with time in diurnally fluctuating temperatures at higher mean temperatures. These qualitative results apply broadly to cases where proteins or other molecules produced in response to high temperatures reduce growth rates. We discuss some of the critical assumptions and predictions of the model and suggest potential extensions and alternatives. Incorporating time-dependent effects will be essential for making more realistic predictions about the physiological and ecological consequences of temperature fluctuations and climate change.

Advances in research of the effect of heat shock proteins on meat quality during postmortem aging

Effect of postmortem aging on meat quality is one of the focuses in the field of meat science,but there are still academic differences about myofibril protein degradation mechanism during postmortem aging. Heat shock proteins( HSPs) can be activated in the condition of ischemia and hypoxia after slaughter. HSPs can affect meat quality during postmortem aging because they can act as chaperone proteins,anti- apoptotic factors,play a role in maintaining protein structure and stability.At present,the impact of HSPs on meat quality,especially of s HSPs,have been widely reported,however,those conclusions are inconsistent. This review mainly describes several important characteristics of HSPs molecular structure and function,and discusses the relationship between HSPs and meat color,tenderness,water holding capacity during postmortem aging.

Combined PI3K/Akt and Hsp90 targeting synergistically suppresses essential functions of alloreactive T cells and increases Tregs.

Acute graft-versus-host disease is still a major cause of transplant-related mortality after allogeneic stem cell transplantation. It requires immunosuppressive treatments that broadly abrogate T cell responses, including beneficial ones directed against tumor cells or infective pathogens. Inhibition of the heat shock protein of 90 kDa has been demonstrated to eliminate tumor cells, as well as alloreactive T cells while preserving antiviral T cell immunity. Here, we show that the suppressive effects of heat shock protein of 90 kDa inhibition on alloreactive T cells were synergistically enhanced by concomitant inhibition of the PI3K/Akt signaling pathway, which is also strongly activated upon allogeneic stimulation. Molecular analyses revealed that this antiproliferative effect was mainly mediated by induction of cell-cycle arrest and apoptosis. In addition, we observed an increased proportion of activated regulatory T cells, which critically contribute to acute graft-versus-host disease control, upon combined heat shock protein of 90 kDa/Akt isoforms 1 and 2 or heat shock protein of 90 kDa/PI3K/p110δ isoform inhibition. Moreover, antiviral T cell immunity was functionally preserved after combined heat shock protein of 90 kDa/Akt isoforms 1 and 2 inhibition. Taken together, our data suggest that the combined heat shock protein of 90 kDa/PI3K/Akt inhibition approach represents a reasonable dual strategy to suppress residual tumor growth and efficiently deplete alloreactive T cells and thus, provide a rationale to prevent and treat acute graft-versus-host disease selectively without impairing pathogen-specific T cell immunity.

Geldanamycin Reduces Aβ-Associated Anxiety and Depression, Concurrent with Autophagy Provocation.

Neurodegenerative disorders are generally characterized by abnormal aggregation and deposition of specific proteins. Amyloid beta (Aβ)-associated neurodegenerative disorder is characterized by an oxidative damage that, in turn, leads to some behavioral changes before the establishment of dementia such as depression and anxiety. In the current study, we investigated the effect of heat shock protein 90 inhibitor geldanamycin (GA) administration 24 h before Aβ injection. In our experiment, 7 days after Aβ injection, elevated plus maze and forced swimming test were conducted to assess anxiety and depression-like behaviors. Levels of autophagy markers and malondialdehyde (MDA) and also activity of catalase in the hippocampus of rats were evaluated. Our behavioral analyses demonstrated that GA pretreatment can significantly decrease anxiety- and depression-like behaviors in Aβ-injected rats. Also, levels of autophagy markers including Atg12, Atg7, and LC3-II increased, while MDA level decreased and the activity of catalase increased in rats pretreated with GA compared to Aβ-injected rats. Thus, we assumed that GA, at least in part, ameliorated Aβ-mediated anxiety and depression by inducing autophagy and improving antioxidant defense system.

Cytokine effects and cellular signaling pathways of grass carp HSP70 in head kidney leukocytes.

In mammals, the cytokine effects of heat shock protein 70 (HSP70) have been well documented. However, the precise role and molecular mechanism of fish HSP70 in cytokine production are poorly understood. In the present study, recombinant grass carp HSP70 (rgcHSP70) was prepared by using the E. coli expression system and its effect on interleukin (IL)-1β (gcil1β) and tumor necrosis factor (TNF)-α (gctnfα) mRNA expression was examined in grass carp head kidney leukocytes (HKLs). Compared with its heated-inactivated protein, rgcHSP70 enhanced the mRNA levels of these two cytokines in a dose- and time-dependent manner. More importantly, the results of enzyme-linked immunoabsorbent assay (ELISA) showed that rgcHSP70 markedly stimulated gcIL-1β and gcTNF-α secretion from the HKLs, for the first time indicating the ability of HSP70 to regulate cytokine release in fish. Moreover, rgcHSP70 displayed more potential to increase the cytokines' release instead of their transcription in the same cell model. Subsequently, the signaling mechanisms governing the cytokine effect of rgcHSP70 were investigated, showing that rgcHSP70-induced gcil1β mRNA expression was suppressed by inhibiting p38 MAPK and NF-κB pathways, while the stimulation of rgcHSP70 on gctnfα mRNA expression were blocked by inhibiting JNK MAPK and NF-κB pathways. In parallel, rgcHSP70 was also shown to activate NF-κB pathway through regulating grass carp IκBα protein levels in a time-dependent oscillation. These findings provide further understanding of cytokine effect and regulatory mechanism of HSP70 in fish and new insights into the roles of HSP70 in teleost immunity.

Effects of the novel heat shock protein 90 inhibitor AUY922 in renal cell carcinoma ACHN and 786-O cells.

The aim of the present study was to investigate the effects of the heat shock protein (HSP)90 inhibitor AUY922 on the proliferation and migratory ability of renal cell carcinoma (RCC) cells. The expression of HSP90 was measured in vitro using western blotting and quantitative polymerase chain reaction in the ACHN and 786-O cell RCC lines, and also in the immortalized normal human proximal tubule epithelium HK-2 cell line. The effects of the time and concentration of AUY922 administration were investigated in the ACHN and 786-O cells, and the cell proliferation was measured using an MTT assay. A Transwell assay was performed to evaluate the migratory ability of ACHN cells following treatment with AUY922 at concentrations of 10, 50 and 100 nM. Western blot analysis and reverse transcription polymerase chain reaction revealed that HSP90 mRNA and protein were overexpressed in the two RCC cell lines compared with the HK-2 cell line. AUY922 inhibited the proliferation of the ACHN and 786-O cells in a time- and concentration-dependent manner, and the migratory ability of the ACHN cells was markedly suppressed subsequent to treatment with AUY922. The present data suggest that the pathogenesis of human RCC may be mediated by HSP90. It was also indicated that the specific HSP90 inhibitor AUY922 plays a therapeutic role in the treatment of RCC, and therefore, HSP90 may be a selective target for molecular-targeted treatments of RCC.

Co-inhibition effect of 17-DMAG and oxaliplatin on proliferation and invasion of colorectal cancer cells

To explore the effect of heat shock protein 90 (HSP90) inhibitor (17-DMAG) and oxaliplatin on the proliferation and invasion of colorectal cancer. After 17-DMAG, oxaliplatin and half-dose combination of 2 drugs processing colorectal cancer SW480 and HCT116 cell lines, CCK8 assay was applied to detect cell viability. RT-PCR and Western blot were used to detect the expression level of the apoptosis-related molecules. Transwell chemokine axis experiment and Western blot were employed to detect cell invasion ability and the expression level of tumor metastasis-associated protein. The growth of SW480 and HCT116 cells was inhibited after the administration of 17-DMAG and oxaliplatin(P<0.05) in dose- and time-dependent manner. Processed by 17-DMAG 100 nmol/L, oxaliplatin 50 mg/L and half-dose combination of 2 drugs, transcription level of the apoptosis inhibitory gene (Bcl-2) in SW480 and HCT116 cells was decreased, the level of apoptosis promoting gene (Bax) transcription and protein PARP-1 spliceosome expression was increased, and the above trend was more obvious when using half-dose combination of 2 drugs. Transwell chemokine axis experiments showed the penetrating relative percentage and expression level of MMP9 and integrin β3 decreased, especially for half-dose combination of 2 drugs. 17-DMAG and oxaliplatin can co-inhibit the proliferation and invasion of colorectal cancer.

Diet and the Anti-inflammatory Effect of Heat Shock Proteins

Stress proteins or heat shock proteins (HSPs) have a critical role in gut health and immune regulation. They have a functional significance as molecular chaperones for cell skeleton proteins and intercellular tight junction proteins. Herewith HSPs ensure gut epithelium integrity and effective intestinal barrier function. In addition, stress protein molecules such as HSP70 are a target for anti-inflammatory regulatory T cells (Tregs). Inflamed sites in the body feature inflammatory-stress induced enhanced levels of HSPs, which enable the immune system to target Tregs selectively to sites of inflammation. We have shown in experimental models of inflammatory diseases that both microbial HSP and endogenous (self) HSP molecules are capable of inducing the expansion of disease suppressive Tregs. Since the gut associated lymphoid tissue (GALT) is well poised towards the induction of regulation and tolerance, we set out to promote HSP expression and induction of Tregs in the gut lymphoid tissues by the oral administration of HSP co-inducing compounds. For the identification, selection and characterization of such compounds we have developed assay systems, such as reporter cell-lines, HSP specific T cell hybridomas and a transgenic mouse model (expression a HSP specific T cell receptor). The introduction of HSP coinducers into the diet constitutes a novel food based preventive or possibly even therapeutic approach in inflammatory diseases.

The HSP90 inhibitor 17-AAG exhibits potent antitumor activity for pheochromocytoma in a xenograft model.

This study aims to investigate the effect of heat shock protein 90 (HSP90) inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG) in the malignant pheochromocytoma using a xenograft mouse model. Treatment with 17-AAG induced a marked reduction in the volume and weight of PC12 pheochromocytoma cell tumor xenografts in mice. Furthermore, 17-AAG also significantly inhibited the expression of HSP90 and its client proteins. Our results validated HSP90 as an important target in pheochromocytoma and provided rationale for the testing of HSP90 inhibitors as a promising therapeutic agent in the antitumor therapy of pheochromocytoma.

Implications of astrocytes in mediating the protective effects of Selective Estrogen Receptor Modulators upon brain damage

Implications of astrocytes in mediating the protective effects of Selective Estrogen Receptor Modulators upon brain damage

Metabolic Reconstruction of Glutamate-Glutamine Cycling: A Flux Balance Approach

Event Abstract Back to Event Metabolic Reconstruction of Glutamate-Glutamine Cycling: A Flux Balance Approach Nelson E. Vega-Vela1, Cynthia M. Jimenez1, George E. Barreto1* and Janneth Gonzalez1 1 Pontificia Universidad Javeriana, Colombia Outline - Astrocyte/Neuron metabolic relationships: A short overview - Metabolic reconstruction: Basic principles - Computational systems biology databases and toolkits: Short review - Metabolic modelling and flux balance analysis of glutamate-glutamine cycle between neurons and astrocytes - Metabolic network analysis and visualization Abstract Cells, tissues, organs and organisms can be understood as a large number of interconnected networks of biochemical pathways, genes, transcripts and proteins which give rise to emergent and specific functions and behaviors in a complex biological system. The complexity exhibited by these systems imposes considerable challenges to understand how they behave and work. For instance, diseases or patho-physiological states are complex conditions that must be considered in a holistic approach because they involve intricate interactions between thousands of components such as genes, transcripts and proteins. In this context, diseases are considered as perturbed states of these networks which can be used to identify components (e.g. metabolites, enzymes, etc.) and interactions that are gained or lost, as well as the biochemical pathways involved. Therefore, discrimination of components and interactions that are not relevant or informative is important to embrace the complexity for understanding biological processes. Systems biology, Network medicine and the recent Systems neuroscience has been successfully applied to make this complexity comprehensible in the different fields of biology, physiology, medicine and neuroscience. To illustrate these approaches within the framework of the workshop “Latin-American School on glial cells in the diseased brain”, we reconstruct the well-known glutamate/glutamine cycle to understand how glutamate can be a neurotoxic agent through glutamate-induced excitotoxicity in neurons and astrocytes. Figure 1 References Barreto GE, Santos-Galindo M, Garcia-Segura LM. Selective estrogen receptor modulators regulate reactive microglia after penetrating brain injury. Front Aging Neurosci, 6:132, 2014. Cabezas R, Ávila M, Gonzalez J, El-Bachá RS, Báez E, Garcia-Segura LM, Jurado Coronel JC, Capani F, Cardona-Gomez GP, Barreto GE. Astrocytic modulation of blood brain barrier: perspectives on Parkinson’s disease. Front. Cell. Neurosci. 8:211, 2014. Barreto GE, White RE, Xu L, Palm CJ, Giffard RG. Effects of heat shock protein 72 (Hsp72) on evolution of astrocyte activation following stroke in the mouse. Exp Neurol. 238(2):284-296, 2012. Barreto G , White RE, Ouyang Y, Xu L, Giffard RG. Astrocytes: targets for neuroprotection in stroke. Cent Nerv Syst Agents Med Chem, 11(2):164-73, 2011. Barreto GE , González J, Torres Y, Morales L. Astrocyte-neuronal crosstalk: Implications for neuroprotection from brain injury. Neurosc Res, 71(2):107-13, 2011. Keywords: astrocyte, Neurons, Metabolic Networks, Flux balance analysis, Neurotoxicity, Computational Biology Conference: Latin-American School on glial cells in the diseased brain (IBRO), Bogotá, Colombia, 13 Jul - 17 Jul, 2015. Presentation Type: Oral Presentation Topic: Neural excitability, synaptic transmission, glia: Cellular and molecular mechanisms Citation: Vega-Vela N, Jimenez CM, Barreto G and Gonzalez J (2015). Metabolic Reconstruction of Glutamate-Glutamine Cycling: A Flux Balance Approach. Conference Abstract: Latin-American School on glial cells in the diseased brain (IBRO). doi: 10.3389/conf.fncel.2015.35.00008 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 22 Feb 2015; Published Online: 16 May 2015. * Correspondence: Prof. George E. Barreto, Pontificia Universidad Javeriana, Bogotá, Colombia, gesbarreto@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Nelson E. Vega-Vela Cynthia M Jimenez George E. Barreto Janneth Gonzalez Google Nelson E. Vega-Vela Cynthia M Jimenez George E. Barreto Janneth Gonzalez Google Scholar Nelson E. Vega-Vela Cynthia M Jimenez George E. Barreto Janneth Gonzalez PubMed Nelson E. Vega-Vela Cynthia M Jimenez George E. Barreto Janneth Gonzalez Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

The synergistic effects of heat shock protein 70 and ginsenoside Rg1 against tert-butyl hydroperoxide damage model in vitro.

Neural stem cells (NSCs) transplanted is one of the hottest research to treat Alzheimer's disease (AD), but cholinergic neurons from stem cells were also susceptible to cell death which Heat shock protein 70 (HSP70) was affirmed to reverse. Related to cognitive impairment, cholinergic nervous cells should be investigated and ginsenoside Rg1 (G-Rg1) was considered to increase them. We chose tert-butyl hydroperoxide (t-BHP) damage model to study in vitro. Functional properties of our recombination plasmid pEGFP-C2-HSP70 were affirmed by SH-SY5Y cells. To opposite the transitory appearance of HSP70, NSCs used as the vectors of HSP70 gene overexpressed HSP70 for at least 7 days in vitro. After transfection for 3 days, G-Rg1 pretreatment for 4 hours, and coculture for 3 days, the expression of acetylcholinesterase (ChAT), synaptophysin, and the ratio of NeuN and GFAP were assessed by western blot; Morphological properties were detected by 3D reconstruction and immunofluorescence. ChAT was markedly improved in the groups contained G-Rg1. In coculture system, the ratio of neurons/astrocytes and the filaments of neurons were increased; apoptosis cells were decreased, compared to monotherapy (P < 0.05). In conclusion, we demonstrated that, as a safe cotreatment affirmed in vitro, overexpression of HSP70 in NSCs plus G-Rg1 promoted nervous cells regeneration from chronic oxidative damage.

Effects of heat shock protein 90 expression on pectoralis major oxidation in broilers exposed to acute heat stress

This study was conducted to determine the effects of heat shock protein 90 (HSP90) expression on pH, lipid peroxidation, heat shock protein 70 (HSP70), and glucocorticoid receptor (GR) expression of pectoralis major in broilers exposed to acute heat stress. In total, 90 male broilers were randomly allocated to 3 groups: control (CON), heat stress (HS), or geldanamycin treatment (GA). On d 41, the broilers in the GA group were injected intraperitoneally with GA (5 μg/kg of BW), and the broilers in the CON and HS groups were injected intraperitoneally with saline. Twenty-four hours later, the broilers in the CON group were moved to environmental chambers controlled at 22°C for 2 h, and the broilers in the HS and GA groups were moved to environmental chambers controlled at 40°C for 2 h. The pH values of the pectoralis major after 30 min and 24 h of chilling after slaughter of HS and GA broilers were significantly lower (P < 0.01) than those of the CON broilers. Heat stress caused significant increases in sera corticosterone and lactic dehydrogenase, the activity of malondialdehyde and superoxide dismutase, the expression of HSP90 and HSP70, and nuclear expression of GR protein in the pectoralis major (P < 0.05). Heat stress induced a significant decrease in GR protein expression in the cytoplasm and GR mRNA expression. Furthermore, the low expression of HSP90 significantly increased levels of lactic dehydrogenase and malondialdehyde and GR protein expression in the cytoplasm under heat stress (P < 0.01), and significantly decreased nuclear GR protein expression (P < 0.01). Heat shock protein 90 was positively correlated with corticosterone and superoxide dismutase activities (P < 0.01), and HSP90 mRNA was negatively correlated with pH after chilling for 24 h. The results demonstrated that HSP90 plays a pivotal role in protecting cells from oxidation.