Hsp70 mRNA Expression Research Articles

Effect of acclimation on thermal limits and hsp70 gene expression of the New Zealand sea urchin Evechinus chloroticus

Seawater temperature is projected to increase globally due to climate change, affecting physiological responses, fitness and survival of marine organisms. Thermal tolerance studies are critical to determine the ability of animals to adapt to future environmental conditions. In this study, we aimed to determine if the thermal limits of the New Zealand Evechinus chloroticus would shift with animal's thermal history. We tested the effect of six thermal regimes on the righting ability, temperature of loss of righting (TLOR), median lethal temperature (LT50), lethal temperature (LT) and the gene expression of the heat shock protein 70 (hsp70) of the New Zealand sea urchin E. chloroticus when exposed to a thermal shock of 1 °C day−1 (duration of 7–16 days depending on the treatment). Treatments consisted of laboratory acclimation for one and four weeks to 18 °C and 24 °C (mean winter (15 °C) and summer temperature (21 °C) + 3 °C of warming, respectively), compared to non-acclimated sea urchins collected during winter (14.6 °C) and summer seasons (20.4 °C). Thermal history did not have a significant effect on the righting ability of E. chloroticus (TLOR ranged between 28 and 29 °C for all treatments) and LT50 (ranged between 29 and 30 °C for all treatments). However, LT of E. chloroticus collected during winter season was significantly lower than animals acclimated for one week at 18 °C. Maximum expression of hsp70 mRNA (Tmax) was observed at around 27–28 °C regardless of treatment; however, relative hsp70 mRNA levels were significantly higher in animals acclimated for four weeks at 24 °C. Despite proving to be a thermotolerant species with LTs around 30 °C, E. chloroticus was unable to increase thermal tolerance and Tmax when acclimated to high temperatures, suggesting that E. chloroticus may have a limited adaptive capacity to modify its phenotype; however, evolutionary adaptations may allow E. chloroticus to adapt to future ocean temperatures.

A mixture of Chinese herbs and a commercial probiotic Bacillus species improves hemato-immunological, stress, and antioxidant parameters, and expression of HSP70 and HIF-1α mRNA to hypoxia, cold, and heat stress in Nile tilapia, Oreochromis niloticus

A 56-day feeding trial was performed to evaluate the effect of herbal-probiotic supplements comprising a mixture of Chinese herbs (CH) (i.e., Astragalus membranaceus, Angelica sinensis, and Crataegus hupehensis) and or commercially prepared probiotic Bacillus species (BS) (i.e., spores of B. subtilis and B.licheniformis) on the physiological response of Nile tilapia. Fish (50 ± 2 g) were fed six diets in duplicates including a control (CT) enriched with BS [at 10 g kg−1], CH [at 10 g kg−1], CHBS1 [CH at 3 g kg−1 and BS at 7 g kg−1], CHBS2 [CH at 5 g kg−1 and BS at 5 g kg−1], and CHBS3 [CH at 7 g kg−1 and BS at 3 g kg−1] for a period of 56 weeks. At the end of the feeding trial, some hemato-immunological parameters and antioxidant indices, as well as stress-related genes in tilapia, were evaluated; and afterwards exposed to hypoxia for 18 h and to cold and heat each for 2 h. The results showed increased hematocrit, red blood cells, haemoglobin, and white blood cells, in fish blood in most of the herbal-probiotic groups compared to the control group (P < 0.05). Lowered cortisol, glucose, as well pyruvate kinase, alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase, and increased myeloperoxidase, total antioxidant capacity, superoxide dismutase, and catalase activities were observed in the serum of all herbal-probiotic groups over the control group (P < 0.05). Besides, all herbal-probiotic groups showed increased expression of heat shock protein 70 and hypoxia-inducible factor 1 alpha mRNAs in the livers of fish (P < 0.05). Among the herbal-probiotic groups, tilapia fed with CHBS2 diet showed enhanced values in most of the indices measured under the different stress conditions (P < 0.05). These findings suggest CHBS2 supplement as a prophylactic against the suppressive effects of stress in tilapia.

Heat shock protein-27 and MiR-17-5p are novel diagnostic and prognostic biomarkers for hepatocellular carcinoma in Egyptian patients

BackgroundHepatocellular carcinoma (HCC) represents the fourth common cause of cancer related death worldwide. HCC prognosis is still miserable due to its late diagnosis and limited treatment strategies, especially in cases with underlying chronic liver diseases as liver cirrhosis (LC). ObjectiveTo check the utility of Hsp27and miR-17-5p expression as early non-invasive diagnostic biomarkers for HCC on top of liver cirrhosis and their association with clinico-pathological parameters. Design and methodsThis case control study included 95 HCC cases and 103 LC patients without HCC. Liver cirrhosis was diagnosed by abdominal ultrasound during clinical examination, liver function tests and liver biopsy. HCC was diagnosed by ultrasound and/or CT or MRI, Alpha Fetoprotein (AFP) and histopathology. Total RNA was extracted from cancerous tissue and surrounding tissues in HCC patients and from blood and serum of both groups. Expression of blood Hsp27 and serum MiR-17-5p were studied by quantitative real time PCR (qRT-PCR). ResultsHsp27 and miR-17-5p mRNA expression were significantly augmented in HCC patients in comparison to LC patients and in malignant tissues as compared to non-malignant adjacent tissues in HCC patients. In blood of HCC patients, Hsp27 mRNA expression was significantly elevated and correlated with big tumour size, number of tumour lesions, high histopathological grades, metastasis and HBV or HCV infection. Serum miR-17-5p mRNA level was significantly enhanced and associated with number of tumour lesions, vascular invasion, stage and metastasis. ConclusionHsp27 and miR17-5p expression levels are augmented in HCC patients and associated with some clinicopathological criteria. Therefore, they can be used as novel non-invasive biomarkers for HCC diagnosis and prognosis in Egyptian patients.

Effects of extracellular Hsp70, lipopolysaccharide and lipoteichoic acid on human monocyte-derived macrophages and differentiated THP-1 cells

Abstract Extracellular Hsp70 (eHsp70) is elevated in blood of patients with inflammatory diseases (e.g. chronic obstructive pulmonary disease), prone to bacterial infections. It stimulates pro-inflammatory effects in cells via TLR2 and TLR4. The main hypothesis was that eHsp70 induces inflammation and alters inflammatory responses to lipopolysaccharide (LPS) and lipoteichoic acid (LTA). We assessed pro-inflammatory cytokines concentrations, TLR2, TLR4 and Hsp70 mRNA expressions, NF-κB and MAPKs activation, and viability after treatment of monocyte-derived macrophages (MDMs) and THP-1 cells with recombinant human (rh) Hsp70, LPS and LTA. rhHsp70, LPS, LTA and their combinations induced IL-6, IL-8 and TNF-α from MDMs. LPS, LTA, and combinations with rhHsp70 increased TLR2 and decreased TLR4 in both cells. rhHsp70 increased TLR2 and TLR4 in THP-1 cells, and TLR2 in MDMs. rhHsp70, LPS and their combinations up-regulated Hsp70 in MDMs, while down-regulated it in THP-1. LPS and its combinations with rhHsp70 lowered cell viability in THP-1 cells and activated caspases-8 and -9. Involvement of NF-κB and MAPKs in inflammatory and survival mechanisms stimulated by rhHsp70 was demonstrated. eHsp70 alone and in combination with bacterial components could provoke inflammation and affect viability of MDMs and THP-1 cells leading to disturbed inflammatory responses in diseases accompanied with bacterial colonisations or infections.

Metformin inhibits cervical cancer cell proliferation by modulating PI3K/Akt-induced major histocompatibility complex class I-related chain A gene expression

BackgroundRecent studies have shown that the classic hypoglycemic drug metformin inhibits tumor growth; however, the underlying mechanism remains unclear. We previously showed that metformin disrupts the sponge effect of long non-coding RNA MALAT1/miR-142-3p to inhibit cervical cancer cell proliferation. In this study, we interrogated the ability of metformin to modulate the anti-tumor immune response in cervical cancer.MethodsThe cell counting kit-8 assay was used to detect the viability of cervical cancer cells. Flow cytometry assays were performed to measure cell apoptosis and cell cycle. Lactate dehydrogenase (LDH) cytotoxicity assay was used to detect NK Cell Cytotoxicity. Relative protein levels were determined by immunoblotting and relative gene levels were determined by quantitative real-time PCR. Tumor Xenograft Modeling was used to evaluate the effect of metformin in vivo.ResultsMetformin inhibited cervical cancer cell proliferation, cervical cancer xenograft growth, expression of PCNA, p-PI3K and p-Akt. Moreover metformin induced cervical cancer cell apoptosis and caused cancer cell cycle arrest. In addition, metformin upregulated the expression of DDR-1 and p53 in human cervical cancer cells. Furthermore, metformin also regulated the mRNA and protein expression of MICA and HSP70 on the surface of human cervical cancer cells via the PI3K/Akt pathway, enhancing NK cell cytotoxicity.ConclusionsIn conclusion, our results suggest that metformin may be used as immunopotentiator to inhibit cervical cancer progression and may be considered a viable candidate for combination therapy with immunotherapy.

Expression of HSP70 mRNA and Protein based on the Thermal Stress in the Primary Hepatocyte Culture of Walleye Pollock (&lt;i&gt;Gadus chalcogrammus&lt;/i&gt;)

Expression of HSP70 mRNA and Protein based on the Thermal Stress in the Primary Hepatocyte Culture of Walleye Pollock (&lt;i&gt;Gadus chalcogrammus&lt;/i&gt;)

Warm Gutta-Percha Techniques Regulate Cell Viability, Heat Shock, and Mineralized Tissue–associated Proteins of Cementoblasts

IntroductionThe aim of this study was to determine the effect of the continuous wave of condensation technique (CWCT) and the thermoplastic gutta-percha injection (TGI) technique on the messenger RNA (mRNA) expressions of heat shock proteins (HSPs) and mineralized tissue–associated proteins of the immortalized mouse cementoblasts (OCCM.30). MethodsCrowns of human premolar teeth with single and straight canals were removed. The root canals were prepared up to the ProTaper Next X5 file (Dentsply Maillefer, Ballaigues, Switzerland) in combination with 2 mL 2.5% sodium hypochlorite solution. Roots (12 ± 2 mm height) were sterilized (121°C for 20 minutes) and placed vertically to the cell culture dishes using a tissue culture insert by opening holes according to the root diameter after the removal of 1 mm from the apex for appropriate adaptation to the petri dish surfaces. Six groups were created: control 1 (without teeth), control 2 (with teeth), AH Plus (Dentsply DeTrey, Konstanz, Germany), single-cone obturation (SC), CWCT, and thermoplastic gutta-percha injection technique (TGI). The viability of the OCCM.30 cells was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability experiments at 24 and 96 hours. The mRNA expression of HSP27, HSP70, and HSP90 and mineralized tissue markers (bone sialoprotein, osteocalcin, runt-related transcription factor 2, type I collagen, and alkaline phosphatase) was evaluated by real-time polymerase chain reaction. ResultsReduced OCCM.30 cell viability was observed in all groups except the control groups. When the SC technique and CWCT and TGI groups were compared, it was observed that heat had a significant negative effect on cell viability (P < .05). A reduction in the mRNA expressions of HSP27, HSP70, and HSP90 was recognized in all test groups when compared with the control 1 group (P < .01). When the warm gutta-percha techniques were compared with the SC technique, a decrease in mRNA expression of HSP27 and HSP90 was noted (P < .01). The HSP70 transcript was similar in the CWCT group and the SC group. Higher HSP70 mRNA expression was observed in the TGI group compared with the SC group. In all groups except the control 1 group, bone sialoprotein, osteocalcin, runt-related transcription factor 2, type I collagen, and alkaline phosphatase mineralized tissue markers were affected, but this negative effect was higher in the heat-treated groups (P < .05). ConclusionsWithin the limitations of this study, it was concluded that warm gutta-percha techniques reduced the mRNA expressions of the genes for HSPs and mineralized tissue–associated proteins of cementoblasts. Further animal studies are needed to clarify the effect of heat on the behavior of cementoblasts histologically in short- and long-term periods.

Living on the upper intertidal mudflat: Different behavioral and physiological responses to high temperature between two sympatric cerithidea snails with divergent habitat-use strategies

Temperature plays a major role in controlling species' distributions, and small-scale variation in the thermal environment are potentially an important factor that governs distributions on a local scale. For untangling the roles of behavioral and physiological adaptations on species' distribution at a small-scale level, we carried out a comparative study of two mudflat snails (genus Cerithidea) by determining these congeners' burying behavior, lethal temperature, cardiac performance and heat-shock protein (hsp70) gene expression. These two sympatric snails occupy different microhabitats on the upper intertidal mudflat. During periods of emersion, C. cingulata inhabits the open mudflat and C. largillierti usually aggregates around small rocks on the upper intertidal mudflat. Our results indicate that the two Cerithidea congeners show different behavioral and physiological responses to high temperature. Compared to C. largillierti, C. cingulata prefers to bury into the mud, has a higher thermal limit and a higher level of inducible expression of hsp70 mRNA, implying important roles of behavioral and physiological adaptations to the harsh thermal environment on the open mudflat. Furthermore, results of generalized additive modelling (GAM) analysis of cardiac performance and coefficient of variation (CV) of hsp70 mRNA expression showed high inter-individual variation in C. cingulata. These results highlight the importance of behavioral and physiological adaptions in sympatric species' distributions on the mudflat and help to shed light on the mechanisms of how small-scale differences in the thermal environment shape sympatric species’ distributions.

Biomarkers of Planarian Dugesia japonica in Response to Herbicide Glyphosate Exposure.

As the worldwide top-selling herbicide, glyphosate is ubiquitously distributed in the natural environment, and its influence on the ecological safety and human health has being increasingly concerned. In this study, mRNA expressions of GPX and three heat shock protein genes in freshwater planarian Dugesia japonica in response to glyphosate were determined, and two oxidative stress parameters were measured. The results suggested that GPX activity can be used as a more sensitive biomarker in contrast with GPX gene expression, and mRNA expressions of Hsp70, Hsp90 genes are more sensitive than Hsp40 for planarians in response to glyphosate stress. Besides, the deduced T-AOC as well as varied GPX activity and mRNA expression levels of Hsps also indicated that glyphosate exposure would inhibit antioxidation and induce oxidative stress in D. japonica, while specific antioxidant systems and stress proteins tried to protect cells by their own regulation. The results of this study will be helpful to elucidate the stress response mechanisms of freshwater planarians to herbicide glyphosate.

Modulatory effect of heat stress on viability of primary cultured chicken satellite cells and expression of heat shock proteins ex vivo

Satellite cells promote muscle repairing and muscle growth. Thereby the intention of the present study was to investigate the beneficial effects of heat stress at different time intervals on chicken satellite cells’ viability. Satellite cells were isolated from 1-day-old chicks and treated at two different temperatures (37 °C and 41 °C) for various time periods (6 h, 12 h, 24 h, 48 h, and 72 h). Both temperatures significantly increased cell viability after 24 h and 48 h. After 12 h, cell viability was significantly increased at 41 °C compared to 37 °C. However, more apoptotic cells were observed at end of the experiment of 41 °C compared to 37 °C. In addition, more live cells were found at early of experimental period at 41 °C than 37 °C. Additionally, protein and mRNA expression of HSP70, HP60 and HSP47 were significantly upregulated throughout the experimental period at temperature of 41 °C compared to those at 37 °C. These results indicate that cell viability and expression of heat stress related proteins/genes are induced by high temperature of 41 °C via heat stress pathway whereas activation of heat stress related proteins/genes are lower at 37 °C. Thus, 41 °C can trigger satellite cells’ viability essential for better cell survival than 37 °C at early incubation time.

Biology behavior of head and neck squamous cell cancer cells changes after knocking down heat shock protein 27

This study aimed to observe the metastatic behavior of head and neck squamous cell carcinoma cells after knocking down heat shock protein (Hsp) 27. The experiment was divided into three groups: the lentivirus vector plasmid of pLenti-shRNA-Hsp27 was transfected into UM-SCC-22B cells as experimental group (shHsp27 group), routine culture of UM-SCC-22B cells as blank control (ctrl group), UM-SCC-22B cells transfection of pLenti-shRNA-ctrl lentivirus vector as negative control (shctrl group). Through real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot assay to detect the mRNA expression of Hsp27 in three groups. MTS assay was performed to detect cell-proliferation changes, wounding healing assay was performed to detect cell-migration changes, and Matrigel Transwell invasion assay was performed to detect cell-invasion changes. The expression of Hsp27 in shHsp27 group decreased signifi-cantly; MTS assay showed that UM-SCC-22B before and after Hsp27 knockdown had similar proliferation rates after being cultured for 24 or 48 h. Compared with the ctrl group, the shHsp27 group decreased the metastatic behavior by 4.38-fold in migration and 2.03-fold in cell invasion. Stably transfected lentivirus vector plasmid of pLenti-shRNA-Hsp27 can efficiently decrease Hsp27 expression and reduce the metastasis ability of UM-SCC-22B.

Love thy neighbor: Social buffering following exposure to an acute thermal stressor in a gregarious fish, the lake sturgeon (Acipenser fulvescens)

Social buffering is a phenomenon where the presence of conspecifics reduces an animal's stress response. Well known in mammals, social buffering was recently described in fishes exhibiting pronounced social hierarchies. Lake sturgeon (Acipenser fulvescens) are a gregarious rather than hierarchical fish. Therefore, we tested their capacity for social buffering following exposure to an acute thermal stress. Isolated or grouped (three or six similarly sized conspecifics) age-0 lake sturgeon were exposed to a critical thermal maximum (CTmax) test. We measured the endocrine and cellular response to acute thermal shock by assessing whole body cortisol concentration and mRNA expression of steroidogenic acute regulatory protein (StAR) and heat shock proteins (hsp90a, hsp90b, and hsp70) during recovery from the CTmax test. Isolation or grouping had no effect on CTmax. Whole body cortisol concentrations in isolated fish were approximately three-fold higher than in grouped fish 1 h post-CTmax and two-fold higher than grouped fish 20 h post-CTmax. Similarly, 1 h post-CTmax, mRNA expression of StAR, hsp90a, hsp90b and hsp70 were three to four-fold higher in isolated fish compared to groups of three and six fish. At 20 h post-CTmax, expression of StAR was approximately two-fold higher in isolated fish, but expression of hsp90a, hsp90b, and hsp70 was not significantly different between isolated and grouped fish. While conspecific presence had no effect on CTmax, the significant reduction of endocrine and cellular stress markers post-CTmax in grouped fish strongly suggests that lake sturgeon may use social buffering to combat potential deleterious effects of exposure to heat stress.

Effect of Glutamine on Antioxidant Capacity and Lipid Peroxidation in the Breast Muscle of Heat-stressed Broilers via Antioxidant Genes and HSP70 Pathway.

Simple SummaryBroilers are heat stress sensitive animals. Heat stress can reduce the meat quality and increase the oxidative stress of broilers. The aim of this study was to investigate the effect of dietary supplementation with Glutamine (Gln) on meat quality, antioxidant capacity, lipid peroxidation, antioxidant genes, and the HSP70 gene and protein expression in broilers exposed to cyclic heat stress. Results showed that Gln improved the meat quality in heat-stressed broilers. In conclusion, our study suggested that dietary Gln supplementation alleviated antioxidant capacity and lipid peroxidation in the breast muscle of heat-stressed broilers by antioxidant genes and the HSP70 pathway.This study investigated whether Glutamine (Gln) could be used as an additive to improve antioxidant capacity in the breast muscle of heat-stressed broilers. Two hundred and forty 22-day-old Arbor Acres broilers in the G1, G2, G3, and G4 groups (n = 60 each) were housed in a cyclic hot environment and fed the basal diet with 0%, 0.5%, 1.0%, and 1.5% Gln, respectively. Compared with the G1 group, dietary 1.5% Gln increased (p < 0.05) pH and b* values, but decreased (p < 0.05) L* cooking loss, drip loss, and water loss rate in breast meat of heat-stressed broilers. Malondialdehyde levels in the breast muscle were lower (p < 0.05) in 1.0% and 1.5% Gln groups than that of the heat-stress group. Compared with the G1 group, dietary 1.5% Gln increased (p < 0.05) catalase (CAT), glutathione, glutathione peroxidase (GSH-Px,) and total antioxidant capacity in the breast muscle of heat-stressed broilers. Furthermore, the CAT, GSH-Px, HSP70 mRNA expression levels, and HSP70 protein expression levels were increased (p < 0.05) in the G3 and G4 groups compared with the G1 group. In sum, Gln alleviated antioxidant capacity and lipid peroxidation in the breast muscle of heat-stressed broilers through antioxidant genes and HSP70 pathways.

MiR-485-5p/HSP90 axis blocks Akt1 phosphorylation to suppress osteosarcoma cell proliferation and migration via PI3K/AKT pathway.

Osteosarcoma (OS) is closely related to the dysregulation of various intracellular signaling pathways, especially the PI3K/Akt signaling pathway. Reportedly, HSP90 was responsible for phospho-Akt stabilization, and both AKT1 and HSP90 were upregulated within osteosarcoma. Herein, we demonstrated that AKT1 and HSP90 mRNA and protein expression were upregulated within osteosarcoma tissues and cells; AKT1 knockdown significantly inhibited OS cell viability. HSP90 knockdown suppressed the phosphorylation of AKT1, decreased ki-67 and Vimentin protein levels, enhanced p21 and E-cadherin protein levels, and inhibited OS cell proliferation and migration; AKT1 overexpression exerted opposing effects and significantly attenuated the effects of HSP90 knockdown. miR-485-5p targeted AKT1 and HSP90 3'-UTR to inhibit AKT1 and HSP90 expression. miR-485-5p overexpression dramatically reduced AKT1, HSP90, and ki-67 proteins, increased E-cadherin protein levels, and inhibited OS cell proliferation and migration. In conclusion, HSP90 knockdown blocked the phosphorylation of AKT1 suppressing the proliferation and migration capacity of OS cells via the PI3K/AKT pathway; miR-485-5p binds to HSP90 and AKT1 in their 3'-UTR to inhibit HSP90 and AKT1 expression, therefore exerting a tumor suppressor function within osteosarcoma.

Chlorpyrifos induces redox imbalance-dependent inflammation in common carp lymphocyte through dysfunction of T-cell receptor γ.

Chlorpyrifos is a poisonous pesticide that is highly toxic to fish and aquatic organisms. However, there are fewer reports about how chlorpyrifos influences the redox balance of immune cells. Herein, the head kidney tissue treated with chlorpyrifos to do transcriptome analysis and TCR γ was screened out. Subsequently, we established TCR γ knockdown and overexpression carp head kidney lymphocyte models, respectively, by using RNA interference and pcDNA3.1. Real-time PCR, fluorescent staining, oxidation and antioxidant kit were used to detect the related factors. We found that TCR γ knockdown significantly increased the mRNA expression of HSP70 and HSP90 and decreased the mRNA expression of SOD and CAT. Meanwhile, TCR γ knockdown reduced the activities of GSH, GSG-PX, T-AOC, CAT and SOD and increased the content of MDA and H2 O2 and activities of iNOS. Adverse results were obtained in TCR γ overexpression group. Additionally, TCR γ knockdown significantly increased the mRNA expression of IFN-γ, IL-1β, IL-8, IL-10, Nrf2 and NF-κB, but relieved TCR γ overexpression, in which the process of inflammation was activated. Our results reported here indicated that chlorpyrifos induces redox imbalance-dependent inflammation in common carp lymphocyte through dysfunction of T-cell receptor γ, and HSPs play potential protective role in entire process.

Curcumin Affects HSP60 Folding Activity and Levels in Neuroblastoma Cells.

The fundamental challenge in fighting cancer is the development of protective agents able to interfere with the classical pathways of malignant transformation, such as extracellular matrix remodeling, epithelial–mesenchymal transition and, alteration of protein homeostasis. In the tumors of the brain, proteotoxic stress represents one of the main triggering agents for cell transformation. Curcumin is a natural compound with anti-inflammatory and anti-cancer properties with promising potential for the development of therapeutic drugs for the treatment of cancer as well as neurodegenerative diseases. Among the mediators of cancer development, HSP60 is a key factor for the maintenance of protein homeostasis and cell survival. High HSP60 levels were correlated, in particular, with cancer development and progression, and for this reason, we investigated the ability of curcumin to affect HSP60 expression, localization, and post-translational modifications using a neuroblastoma cell line. We have also looked at the ability of curcumin to interfere with the HSP60/HSP10 folding machinery. The cells were treated with 6, 12.5, and 25 µM of curcumin for 24 h, and the flow cytometry analysis showed that the compound induced apoptosis in a dose-dependent manner with a higher percentage of apoptotic cells at 25 µM. This dose of curcumin-induced a decrease in HSP60 protein levels and an upregulation of HSP60 mRNA expression. Moreover, 25 µM of curcumin reduced HSP60 ubiquitination and nitration, and the chaperonin levels were higher in the culture media compared with the untreated cells. Furthermore, curcumin at the same dose was able to favor HSP60 folding activity. The reduction of HSP60 levels, together with the increase in its folding activity and the secretion in the media led to the supposition that curcumin might interfere with cancer progression with a protective mechanism involving the chaperonin.

Effects of luteolin on treatment of psoriasis by repressing HSP90

AimsThis study was conducted to further clarify the efficacy and potential of luteolin in treating psoriasis and to explore its inner mechanisms. MethodsA pharmacology network displayed the construction of a drug disease target prediction method. The prediction technique was validated via cell experiments in vitro and animal experiments in vivo, respectively. The effects of IFN-γ and luteolin were detected in HaCaT cells. The secretion of exosome and expression of mRNA and protein were detected to explain the relationship between luteolin's regulation of HSP90 (HSP90α and HSP90β) activity in vitro. An in vivo psoriasis mouse model was established to further explore the efficacy of luteolin. Morphological and histological changes in skin lesions were observed, and the CD63, calnexin, Hsp90α, and Hsp90β protein expression was analyzed. Peripheral blood mononuclear cells (PBMCs) were separated and detected via flow pattern analysis to determine how luteolin effects the immune cells in a psoriasis model. ResultsLuteolin as a candidate compound is predicted to have a molecular-target correspondence with HSP90 according to a pharmacology network analysis. Cell experiments indicated that the pathogenesis of psoriasis was significantly related to the increase in IFN-γ, which promoted the transcriptional expression and exosome secretion of HSP90 in HaCaT cells; conversely, luteolin inhibited those and alleviated the promotion of IFN-γ. The effect of luteolin on HSP90 was slightly weaker than that of INF-γ. Animal experiments indicated that the efficacy of luteolin was similar to that of 17-AAG, which both alleviated skin tissue lesions and symptoms, improved the expression of Hsp90 mRNA and protein in skin tissue, and promoted exosome secretion of Hsp90 in plasma. For immune cells in mice with psoriasis, luteolin reduced the proportion of Th1/Th2 and Th17/Treg and inhibited the increase in Th1 and Th17 in the peripheral blood. ConclusionLuteolin can relieve the lesions and symptoms of psoriasis through reversing the effects of IFN-γ, inhibiting the expression and exosome secretion of HSP90, and regulating the proportion of immunocytes. Therefore, this study provides the possible mechanisms and potential utilization of luteolin as a novel treatment for psoriasis.

Curcumin Alleviates IUGR Jejunum Damage by Increasing Antioxidant Capacity through Nrf2/Keap1 Pathway in Growing Pigs.

Simple SummaryIntrauterine growth retardation (IUGR) is usually defined as fetal growth below the 10th percentile for gestational age and results in impaired growth and development of the fetus and/or its organs during gestation. IUGR not only has a permanent effect on postnatal growth and long-term health, but also results in high fetal mortality and morbidity. Recent results have demonstrated that IUGR can cause jejunum damage in piglets. The jejunum is not only the main organ for the digestion and absorption of nutrients, but also an immune organ in the body. However, few effective methods to alleviate jejunum damage and oxidative stress in IUGR pigs have been found. In recent years, studies have found that curcumin (CUR) may be an effective and safe feed additive for regulating oxidative stress in the body. Our results indicated that dietary added 200 mg/kg curcumin to the basal diet can increase the antioxidant capacity of the IUGR growing pigs, jejunum and alleviate the damage in jejunum of the IUGR growing pigs. Therefore, the use of curcumin as a feed additive has certain economic value.The purpose of this study was to explore the effects of curcumin on IUGR jejunum damage. A total of 24 IUGR and 12 normal-birth weight (NBW) female crossbred (Duroc × Landrace × Large White) piglets were randomly assigned into three groups at weaning (26 days): IUGR group, NBW group, and IUGR + CUR group, which were fed diets containing 0 mg/kg (NBW), 0 mg/kg (IUGR) and 200 mg/kg (IUGR + CUR) curcumin from 26 to 115 days of age. Results showed that dietary supplementation with 200 mg/kg curcumin significantly increased the total superoxide dismutase (T-SOD) activity and decreased the malondialdehyde (MDA) content in the jejunum of IUGR pigs (p < 0.05). Results of real-time PCR showed that the IUGR + CUR group significantly increased the gene expression of NF-E2-related factor 2 (Nrf2) (p < 0.05), and increased the glutamate-cysteine ligase catalytic subunit (GCLC), superoxide dismutase 1 (SOD1), glutamate-cysteine ligase modifier subunit (GCLM), and NAD(P)H quinone dehydrogenase 1 (NQO1) mRNA expression compared with the IUGR group (p < 0.05). Western blot results showed that dietary supplementation with 200 mg/kg curcumin significantly increased the protein levels of Nrf2 and NQO1. Compared with the IUGR group, pigs in IUGR + CUR group showed significantly decreased the levels of tumor necrosis factor-α (TNFα), interleukin-6 (IL-6), and interferon gamma (IFNγ) (p < 0.05), and increased the interleukin-2 (IL-2) level (p < 0.05). Dietary supplementation with 200 mg/kg curcumin significantly reduced cysteinyl aspartate specific proteinase 3 (caspase3), BCL2-associated X protein (bax), B-cellCLL/lymphoma 2 (bcl2), and heat-shock protein 70 (hsp70) mRNA expression, and increased occludin (ocln) mRNA expression (p < 0.05). In conclusion, dietary supplementation with 200 mg/kg curcumin can alleviate jejunum damage in IUGR growing pigs, through Nrf2/Keap1 pathway.

Effects of stocking density on the growth performance, serum biochemistry, muscle composition and HSP70 gene expression of juvenile golden pompano Trachinotus ovatus (Linnaeus, 1758)

Golden pompano (Trachinotus ovatus) is one of the most important cultured marine fish in Southeast Asia. Stocking density affects the growth performance and welfare of fish in aquaculture. A 70-day rearing experiment was performed to evaluate the effect of different stocking densities on the growth performance, feed conversion ratio, serum biochemistry, muscle quality, antioxidant capacity and HSP70 mRNA expression of juvenile golden pompano cultured in off-shore sea cages. The experimental design was completely randomized using three replications with three treatments: low (100 fish/m3), medium (200 fish/m3), and high (300 fish/m3) stocking densities. Fish growth decreased significantly, and the feed conversion ratio increased significantly with increasing stocking density (P < .05). The serum glucose concentration increased significantly (P < .05), and the total protein, total triglyceride and total cholesterol concentrations were significantly reduced with increasing stocking density (P < .05). High stocking density led to elevated serum cortisol levels and decreased thyroxine levels. There was a significant decrease in the muscular crude fat content with increasing stocking density (P < .05), while the moisture, ash and protein contents did not significantly differ among the groups (P > .05). In this experiment, superoxide dismutase, glutamine transaminase and total antioxidant capacity increased significantly with increasing stocking density (P < .05). The expression levels of heat shock protein 70 were significantly up-regulated in the liver, kidney and brain as stocking density increased (P < .05); however, there was no significant difference in the spleen (P > .05). Overall, high stocking density was a chronic stressor in this experiment and had a negative effect on the growth, feed conversion ratio and animal welfare of T. ovatus.

Hsp70s transcription-translation relationship depends on the heat shock temperature in zebrafish

Virtually all organisms respond to heat shock by transcription of genes encoding for heat shock proteins (HSPs), but the mechanisms behind post-transcriptional regulation are not known in detail. When we exposed zebrafish to 5 and 7 °C above normal rearing temperature for 30 min, hsp70 mRNA expression was 28 and 150 -fold higher than in control, respectively. Protein expression, on the other hand, showed no significant change at the +5 °C and a 2-fold increase at the +7 °C exposure. This suggests that the transcription of hsp70 gene does not immediately correspond to translation to related proteins under certain stress temperatures, but, when the temperature is higher, and potentially detrimental, transcription and translation are intimately coupled. Those results confirm that temperature is an important abiotic factor involved in heat shock post-transcriptional regulation mechanisms in fish. However, further studies are needed to determine the relationship between this environmental factor and post-transcriptional regulation mechanisms. Earlier, the coupling/uncoupling of hsp transcription and translation has only been studied using cold-water fish, or zebrafish embryos. With current findings, we suggest this mechanism might be present even in adult warm water fish like the zebrafish.