Levels Of Heat Shock Protein Research Articles

Phytic acid-Cu2+ framework/Cu2-xS nanocomposites with heat-shock protein down-modulation ability for enhanced multimodal combination therapy

Mild-temperature photothermal therapy (mPTT) has shown some advantages over traditional photothermal therapy, such as reducing the damage to surrounding healthy tissues and minimizing side effects. Nevertheless, cancer cells can easily repair damage caused by mild hyperthermia due to heat shock proteins (HSPs). Thus, it is imperative to maximize the mPTT efficiency by down-regulating HSPs overexpression and combining other cancer treatments. Herein, we report the synthesis of phytic acid (PA)-Cu2+ framework/copper sulfide (Cu2-xS) nanocomposites (abbreviated as PA-Cu/Cu2-xS NPs) as the novel therapeutic platform that can down-regulate HSPs overexpression for enhanced multimodal mPTT/chemodynamic therapy (CDT)/chemotherapy. PA-Cu/Cu2-xS NPs were prepared through self-assembly and in-situ vulcanization strategy, resulting in irregular-shaped particles with an approximate size of 100 nm. PA-Cu/Cu2-xS NPs display a plasmon effect from Cu2-xS, which enhances near-infrared (NIR) absorption and possesses excellent photothermal conversion efficiency (41.7%). Moreover, PA-Cu/Cu2-xS NPs exhibit Fenton-like reaction activity resulting from the Cu ions for CDT, and the reaction activity can be further improved 1.3 times due to mild hyperthermia during mPTT. Furthermore, the generated hydroxyl radical (•OH) can effectively decrease HSPs level to enhance mPTT. PA-Cu/Cu2-xS NPs can also serve as a drug delivery system, and they are capable of loading doxorubicin (DOX) with a loading ability (20.7%). Combining mPTT/CDT/chemotherapy exhibits significant inhibition of tumor growth. This approach can serve as a basis for designing more exquisite platforms that combine mPTT with other therapies to achieve more effective cancer treatment.

Thioredoxin-1 inhibits the activation of IRE1 by targeting Hsp90/p-Cdc37 chaperone complex in Parkinson disease.

Endoplasmic reticulum stress is implicated in the etiopathogenesis of Parkinson disease (PD). Our previous study has revealed that thioredoxin-1 (Trx-1) attenuated IRE1 activation in 1-methyl-4-phenylpyridinium ion (MPP+)/1-methy-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD models. However, its exact mechanism has been largely unclear. In this research, it was reported for the first time that the protein levels of heat shock protein 90 (Hsp90) and phosphorylated cell division cycle 37 (p-Cdc37) were significantly decreased and the interaction of Hsp90/p-Cdc37 complex with IRE1 was disturbed in MPP+/MPTP-induced PD models. Trx-1 overexpression reversed the expression of Hsp90 and p-Cdc37 in cultured cells and the substantia nigra pars compacta of mice. More importantly, Trx-1 overexpression enhanced the interaction of Hsp90/p-Cdc37 complex with IRE1. In conclusion, our data demonstrated that Trx-1 inhibited IRE1 activation in PD by elevating the expression of Hsp90 and p-Cdc37 and strengthening the interaction of Hsp90/p-Cdc37 complex and IRE1.

Elevated HSP70 and HSP90 as Predictive Markers of Immune Activation and Lung Injury in SARS-COV-2 Disease.

Heat shock proteins (HSPs) are involved in innate and adaptive immune responses, especially inflammatory responses due to immune cell activation. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was one of the most important causes of death in the recent pandemic. Increased cellular stress and excessive inflammation are common in coronavirus disease-19 (COVID-19), although the underlying mechanisms are still poorly understood. To evaluate the relationship between HSP and the pathological effects of COVID-19. A group of 107 patients was categorized to two populations (mild and severe) based on their chest high-resolution computed tomography (HRCT) results. The HSP70, HSP90 alpha, and serum levels of C-reactive protein (CRP) were measured by enzyme-linked immunosorbent assay (ELISA). Lactate dehydrogenase (LDH), and creatine phosphokinase (CPK) were measured by the automated analyzer. Our data showed increased levels of HSP70 and HSP90 in patients with COVID-19. The HSPs levels were elevated in the severe group compared to the mild group. This study demonstrated a positive correlation between both elevated levels of HSP70, HSP90, and HRCT grade and also a positive correlation with CRP and CPK in the severe group. HSP90 and HSP70 contribute to excessive immune responses and cytokine storms. They may serve as prognostic serum markers for COVID-19 lung injury. Additionally, they are candidates for anti-inflammatory therapy.

Electric Field Stimulation of Stem Cell Culture Media Increases Oxygen Radical Uptake Capacity

BACKGROUND: Stem cell culture medium is garnering attention because it comprises growth factors and exosomes secreted by stem cells. In this study, stem cell culture medium was subjected to electric field stimulation using the cells alive system (CAS) to determine the antiaging effects of this medium on dermal fibroblasts.METHODS: The medium was added to the monolayer cultures of human fibroblasts with or without 100 μg/mL advanced glycation end products (AGE), and the cells were incubated for 4 hours. Total RNA was extracted, and mRNA levels of several AGE receptors and heat shock proteins were measured using quantitative reverse transcriptase-polymerase chain rection (qRT-PCR).RESULTS: Similar amounts of exosomes were extracted from the supernatants of stem cells cultured with (CAS+) and without (CAS−) electric field stimulation. The results demonstrated that CAS+ had a higher oxygen radical absorbance capacity than CAS−, indicating its high antioxidant activity. Furthermore, the expression of domain-containing link scavenger receptor-1 mRNA, which is involved in the degradation of cell surface receptors for AGE under glycation stress, significantly increased. The electric field stimulation also increased the expression of several heat shock proteins, which play important roles in cellular responses generated against glycation stress. The electric field stimulation of the stem cell culture medium effectively promoted the endocytosis and degradation of AGE, thereby exerting antioxidant effects and inducing glycation stress.CONCLUSION: Electric field stimulation of stem cell culture medium enhanced the expression of Hsp and its antioxidant activity in fibroblasts.KEYWORDS: AGE, electric field stimulation, exosome, fibroblast, oxygen radical absorbance capacity, stem cell

Biological responses of stellate sturgeon fingerlings (Acipenser stellatus) immersed in HSP inducer to salinity changes

Changes in salinity is a stressful and energy-consuming process in fish which give rise to mortalities, especially in fish fingerlings that are more sensitive during the early stages of their life. In the present study, the effects of three salinities, 3‰ (downstream of river), 8‰ (estuarine), and 13‰ (the maximum salinity in the Caspian Sea), on HSP70 gene expression, cortisol level, immune response (lysozyme, complement C3, IgM), and antioxidant enzyme activities (SOD, CAT, T-AOC) of the stellate sturgeon fingerlings in the presence of HSP inducer compound (TEX-OE®) were evaluated. Our results showed that levels of plasma cortisol and heat shock protein (HSP70) in Acipenser stellatus fingerlings increased due to salinity changes. In the presence of the HSP inducer, HSP70 expression in both gill and liver was significantly increased, whereas cortisol level was notably decreased. Exposure to salinity changes resulted in an increase in antioxidant defense activities (SOD, CAT, and T-AOC) and immune response (lysozyme, IgM, and C3) in the presence of an HSP inducer. In conclusion, an HSP-inducing compounds can have a positive effect in strengthening the immunity and antioxidant system of sturgeon fingerlings by increasing the expression of the HSP70 gene against salinity fluctuations and generally increase the body's physiological tolerance.

Exploring the impact of temporal heat stress on skeletal muscle hypertrophy in bovine myocytes

The primary aim of this investigation was to explore the impact of different temporal stress conditions on the regulators associated with skeletal muscle hypertrophy in bovine myocytes. Bovine satellite cells (BSCs) were extracted from three-month-old Holstein bull calves and subjected to myogenic differentiation under three thermal treatments: 38 °C (control; CON), 39.5 °C (moderate heat stress; MHS), and 41 °C (extreme heat stress; EHS) for a duration of 3 or 48 h. Exposure to EHS resulted in elevated (P < 0.01) expression levels of heat shock protein (HSP)20, HSP27, HSP70, and HSP90, along with increased (P < 0.01) protein levels. Moreover, cells exposed to MHS and EHS exhibited enhanced (P < 0.01) gene expression of myoblast determination protein 1 (MyoD), while myogenin (MyoG) was overexpressed (P < 0.01) in cells exposed to EHS. These findings suggest that heat exposure can potentially induce myogenic differentiation through the modulation of myogenic regulatory factors. Furthermore, our investigations revealed that exposure to EHS upregulated (P < 0.01) myosin heavy chain (MHC) I expression, whereas MHC IIA (P < 0.01) and IIX (P < 0.01) expression were increased; P < 0.01) under MHS conditions. These observations suggest that the temperature of the muscle may alter the proportion of muscle fiber types. Additionally, our data indicated that EHS activated (P < 0.01) the expression of insulin-like growth factor 1 (IGF-1) and triggered the activation of the Akt/mTOR/S6KB1 pathway, a known anabolic pathway associated with cellular protein synthesis. Consequently, these altered signaling pathways contributed to enhanced protein synthesis and increased myotube size. Overall, the results obtained from our current study revealed that extreme heat exposure (41 °C) may promote skeletal muscle hypertrophy by regulating myogenic regulatory factors and IGF-1-mediated mTOR pathway in bovine myocytes.

Lactobacillus johnsonii N5 from heat stress-resistant pigs improves gut mucosal immunity and barrier in dextran sodium sulfate-induced colitis

Developing effective strategies to prevent diarrhea and associated-gut disorders in mammals has gained great significance. Owing to the many health benefits provided by the commensal microbiota of the intestinal tract, such as against environmental perturbation, we explored the host phenotype-associated microbes and their probiotic potential. Based on the observations that the chronic heat stress-exposed weaned piglets present as heat stress-susceptible (HS-SUS) or heat stress-resistant (HS-RES) individuals, we confirmed the phenotypic difference between the two on growth performance (P < 0.05), diarrhea index (P < 0.001), intestinal heat shock protein 70 (HSP70) regulation (P < 0.01), and inflammatory responses (P < 0.01). By comparing the gut microbiome using 16S rRNA gene sequencing and KEGG functional analysis, we found that Lactobacillus johnsonii exhibited significantly higher relative abundance in the HS-RES piglets than in the HS-SUS ones (P < 0.05). Further experiments using a mouse model for chemical-induced inflammation and intestinal injury demonstrated that oral administration of a representative L. johnsonii N5 (isolated from the HS-RES piglets) ameliorated the clinical and histological signs of colitis while suppressing intestinal pro-inflammatory cytokines TNF-α and IL-6 production (P < 0.05). We found that N5 treatment enhanced tight junction proteins ZO-1 and occludin and cytoprotective HSP70 levels under physiological condition and restored their mucosal expressions in colitis (P < 0.05). In support of the high production of the anti-inflammatory cytokine IL-10, N5 promoted the intestinal Peyer's patches MHCII+ and CD103+ dendritic cell populations (P < 0.05), increased the regulatory T (Treg) cell numbers (P < 0.05), and decreased the Th17 population and its IL-17a production under physiological condition and during colitis (P < 0.01). Our results shed light on understanding the interaction between commensal Lactobacillus and the host health, and provide L. johnsonii N5 as an alternative to antibiotics for preventing diarrhea and intestinal diseases.

Anti-HSP70 alleviates cell migration and proliferation in colorectal cancer cells (CRC) by targeting CXCR4 (in vitro study).

Colorectal cancer (CRC), the third most common cancer in men and women, accounts for 8% of new cancer cases in the US and 8 to 9% of the nation's anticipated cancer mortality in 2014. In the pathophysiology of colon cancer, heat shock protein 70 (HSP70) and CXCR4 are essential. In this research, we concentrated on the connection between CXCR4 expression and HSP70 inhibitor activity in the development of colorectal cancer. The HSP70 inhibitor's effect on cell proliferation was also evaluated. Samples were obtained from patients with CRC; the surrounding marginal tissues were considered healthy. The One CRC cell lines (HCA-7) were divided into two groups based on untreated and treated with anti-HSP70. HSP70 and CXCR4 mRNA expression and migration (Wound healing assay) were measured in these groups. Also, we evaluated the expression levels of HSP70 and CXCR4 in thirty CRC and healthy non-cancerous samples (Using Real-time PCR and Western Blotting). Moreover, we examined the viability of CRC cells in untreated and treated groups with anti-HSP70. Higher expression levels of CXCR4 (p < 0.0001) and HSP70 (p = 0.002) mRNA were observed in patients who had CRC. In contrast, lower mRNA expressions of HSP70 (p < 0.0001) and CXCR4 (P < 0.0001) were detected in the CRC cell line (HCA-7) after being treated with anti-HSP70. Moreover, the viability and migration of cancer cells were remarkably reduced in CRC cells treated with anti-HSP70. Our study's innovation was the in vitro demonstration of inhibiting HSP70 in the CRC cancer cell line drastically reduced CXCR4 expression, viability, and cancer cell migration. These findings may pave the way for additional studies on CRC cancer treatment and be examined in vivo in studies, given that the primary goal of therapy is to decrease the viability and spread of cancer cells.

A Review on the Involvement of Heat Shock Proteins (Extrinsic Chaperones) in Response to Stress Conditions in Aquatic Organisms.

Heat shock proteins (HSPs) encompass both extrinsic chaperones and stress proteins. These proteins, with molecular weights ranging from 14 to 120 kDa, are conserved across all living organisms and are expressed in response to stress. The upregulation of specific genes triggers the synthesis of HSPs, facilitated by the interaction between heat shock factors and gene promoter regions. Notably, HSPs function as chaperones or helper molecules in various cellular processes involving lipids and proteins, and their upregulation is not limited to heat-induced stress but also occurs in response to anoxia, acidosis, hypoxia, toxins, ischemia, protein breakdown, and microbial infection. HSPs play a vital role in regulating protein synthesis in cells. They assist in the folding and assembly of other cellular proteins, primarily through HSP families such as HSP70 and HSP90. Additionally, the process of the folding, translocation, and aggregation of proteins is governed by the dynamic partitioning facilitated by HSPs throughout the cell. Beyond their involvement in protein metabolism, HSPs also exert a significant influence on apoptosis, the immune system, and various characteristics of inflammation. The immunity of aquatic organisms, including shrimp, fish, and shellfish, relies heavily on the development of inflammation, as well as non-specific and specific immune responses to viral and bacterial infections. Recent advancements in aquatic research have demonstrated that the HSP levels in populations of fish, shrimp, and shellfish can be increased through non-traumatic means such as water or oral administration of HSP stimulants, exogenous HSPs, and heat induction. These methods have proven useful in reducing physical stress and trauma, while also facilitating sustainable husbandry practices such as vaccination and transportation, thereby offering health benefits. Hence, the present review discusses the importance of HSPs in different tissues in aquatic organisms (fish, shrimp), and their expression levels during pathogen invasion; this gives new insights into the significance of HSPs in invertebrates.

Dietary selenium nanoparticles positively modulate the growth and immunity of seabream (Sparus aurata) fingerlings exposed to low salinity stress and Vibrio parahaemolyticus challenge

This study aimed to investigate the effects of selenium nanoparticle (Nano-Se) supplementation on the growth performance, physiological parameters, antioxidant system, and immune response of seabream (Sparus aurata) under normal conditions, as well as after low salinity stress and challenge with Vibrio parahaemolyticus. The fish were divided into five experimental groups that received experimental diets supplemented with different levels of Nano-Se (0, 0.125, 0.25, 0.5, and 0.75 mg/kg Nano-Se) for 14 weeks. The results revealed that Nano-Se supplementation had a positive impact on growth performance, including final weight, weight gain, daily weight gain, specific growth rate, and feed conversion ratio. Nano-Se significantly increased the expression of insulin-like growth factors 1 and 2. The findings suggest that Nano-Se promotes growth in seabream under both normal and stressful conditions. Low salinity stress and Vibrio parahaemolyticus challenge were found to increase the levels of serum cortisol and the heterophil and lymphocyte (H/L) ratio while decreasing the population of lymphocytes, white blood cells, and immunoglobulin M. However, Nano-Se supplementation mitigated these effects. Nano-Se exhibited a protective effect by restoring the H/L ratio to normal levels and enhancing the immune response. Our expression analyses indicated a significant interaction between Nano-Se and stress. Nano-Se reduced the upregulated levels of heat shock protein 70 and interleukin 1β caused by low salinity and bacterial challenge, bringing them closer to normal values, compared to the groups without Nano-Se supplementation. Nano-Se slightly increased the mRNA levels of interleukin 1β under healthy conditions, particularly at concentrations of 0.5 and 0.75 mg/kg. Additionally, Nano-Se improved the serum biochemical profiles and carcass composition of seabream. Notably, the concentrations of 0.5 and 0.75 mg/kg increased the survivability of seabream exposed to low salinity stress and V. parahaemolyticus challenge. These findings suggest that Nano-Se can be effectively utilized to enhance the health and performance of seabream in aquafarms.

The Use of Low-Level Laser Therapy (LL LT) in the Symptomatic Treatment of Vulvar Cancer IV st.

Cutaneous squamous cell carcinoma (CSCC) is a malignant tumor originating from keratinocytes in the epidermis, which seriously affects people's life safety. Heat shock proteins (HSPs) are molecular chaperones that help maintain protein stability and are involved in tumor activities. However, there is currently no systematic study of HSPs in CSCC. In this study, we used qPCR and WB to explore the expression of 14 HSPs at mRNA and protein levels in human immortalized keratinocytes HACAT cells, human CSCC cell lines COLO-16 and SCL-1 cells, and collected human CSCC samples for histological verification. Eventually, we found that the expression levels of different HSPs in CSCC were different. HSPB6, HSPB8 and HSPB11 showed no dysregulation in CSCC. HSPB4, HSPB5, HSPB7, HSPB10 and HSP40 were significantly upregulated in CSCC cells, while HSPB1, HSPB9, HSP60, HSP70, HSP90 and HSP100 were significantly downregulated. In summary, we validated the expression of 14 common HSPs in CSCC, which may lay the foundation for the study of HSPs in CSCC.

Differential expression of heat shock proteins in cutaneous squamous cell carcinoma

Cutaneous squamous cell carcinoma (CSCC) is a malignant tumor originating from keratinocytes in the epidermis, which seriously affects people's life safety. Heat shock proteins (HSPs) are molecular chaperones that help maintain protein stability and are involved in tumor activities. However, there is currently no systematic study of HSPs in CSCC. In this study, we used qPCR and WB to explore the expression of 14 HSPs at mRNA and protein levels in human immortalized keratinocytes HACAT cells, human CSCC cell lines COLO-16 and SCL-1 cells, and collected human CSCC samples for histological verification. Eventually, we found that the expression levels of different HSPs in CSCC were different. HSPB6, HSPB8 and HSPB11 showed no dysregulation in CSCC. HSPB4, HSPB5, HSPB7, HSPB10 and HSP40 were significantly upregulated in CSCC cells, while HSPB1, HSPB9, HSP60, HSP70, HSP90 and HSP100 were significantly downregulated. In summary, we validated the expression of 14 common HSPs in CSCC, which may lay the foundation for the study of HSPs in CSCC.

The role of protein kinase R in placental inflammation, mtUPR and apoptosis

IntroductionPlacental inflammation is implicated in the pathophysiology of many pregnancy complications, including fetal growth restriction, preeclampsia, gestational diabetes, and choriocarcinoma. Mitochondrial dysfunction, one of the outcomes of placental inflammation, is characterized by loss of membrane potential, accumulation of oxygen radicals, mitochondrial protein folding defects, and disturbances in mitochondrial dynamics. Protein kinase R (PKR) is stimulated by double-stranded RNA and bacterial endotoxins in the presence of pathogens and is a critical immune response enzyme. PKR is also correlated with the cell death response during endoplasmic reticulum stress. In this study, we aim to investigate the effects of PKR activity stimulated by lipopolysaccharide (LPS), and double-stranded RNA analog (Poly I:C) on mitochondrial unfolded protein response (mtUPR), mitochondrial membrane potential, apoptosis, and oxidative stress in placental trophoblasts. MethodsWe applied LPS and Poly I:C to BeWo cells to induce PKR activation. In addition, cells were treated with 2-aminopurine (2-AP) to inhibit the kinase activity of PKR. Protein levels of ATP-dependent Clp protease proteolytic subunit (CLPP) and heat shock protein 60 (HSP60) were determined after treatments. Apoptotic markers were detected by real-time PCR and flow cytometry. PKR-induced reactive oxygen radicals (ROS) accumulation and mitochondrial membrane potential change were assessed by flow cytometry. ResultsIt was determined that PKR activation-induced apoptosis in BeWo cells by reducing the levels of mtUPR proteins (CLPP and HSP60) and caused a decrease in mitochondrial membrane potential. PKR inhibition was sufficient for decreases in apoptotic markers and caused a reduction in the ratio of depolarized and ROS (+) cells. DiscussionOur results showed that LPS and Poly I:C administration stimulated PKR in BeWo cells in vitro. Furthermore, PKR activation is correlated with the levels of proteins involved in mitochondrial homeostasis and apoptosis. Our findings will contribute to understanding the role of PKR activation in placental inflammation and related diseases.

The protective role of tetraploidy and nanoparticles in arsenic-stressed rice: Evidence from RNA sequencing, ultrastructural and physiological studies

Genome doubling in plants induces physiological and molecular changes to withstand environmental stress. Diploid rice (D-2x) and its tetraploid (T-4x) plants were treated with 25 μM Arsenic (As) and 15 mg L−1 TiO2 nanoparticles (NPs), and results indicated decreased growth and photosynthetic activity with high accumulation of reactive oxygen species (ROS) due to the As-toxicity in rice lines, significantly in D-2x rice plants. The treatment of As-contaminated rice with TiO2 NPs resulted in increased root length (8.17%) and chlorophyll AB (13.28%) and decreased electrolyte leakage (21.76%) and H2O2 (17.65%) contents than its counterpart diploid rice. Moreover, TiO2 NPs improved the activity of peroxidase, catalase, glutathione, and superoxide dismutase and reduced lipid peroxidation due to lower ROS production in D-2x and T-4x under As toxicity. Transcriptome analysis revealed abrupt changes in the expression levels of key signaling heat shock proteins, tubulin, aquaporins, As, and metal transporters under As toxicity in T-4x and D-2x lines. The KEGG and GO studies highlighted the striking distinctions between rice lines under As-stress in glutathione metabolism, H2O2 catabolic process, MAPK signaling pathway, and carotenoid biosynthesis terms, revealing consistency between physiological and molecular results. Root cells from D-2x rice were significantly more distorted by As poisoning than those from 4x rice, and cell organelles, such as mitochondria and endoplasmic reticulum, were changed or deformed. These findings proved the superiority of tetraploid rice lines over their diploid counterpart in coping with As-stress.

The Role of Hyperthermia in Potentiation of Anti-Angiogenic Effect of Cisplatin and Resveratrol in Mice Bearing Solid Form of Ehrlich Ascites Tumour

The aim of this study was to investigate the therapeutic potential of resveratrol in combination with cisplatin on the inhibition of tumour angiogenesis, growth, and macrophage polarization in mice bearing the solid form of an Ehrlich ascites tumour (EAT) that were exposed to whole-body hyperthermia treatment. In addition, we investigated whether a multimodal approach with hyperthermia and resveratrol could abolish cisplatin resistance in tumour cells through the modulation of histone deacetylase (HDAC) activity and levels of heat shock proteins (HSP70/HSP90) and contribute to the direct toxicity of cisplatin on tumour cells. The tumour was induced by injecting 1 × 106 EAT cells subcutaneously (sc) into the thighs of Balb/c mice. The mice were treated with resveratrol per os for five consecutive days beginning on day 2 after tumour injection and/or by injecting cisplatin intraperitoneally (ip) at a dose of 2.5 mg/kg on days 10 and 12 and at a dose of 5 mg/kg on day 15. Immediately thereafter, the mice were exposed to systemic hyperthermia for 15 min at a temperature of 41 °C. The obtained results showed that the administration of resveratrol did not significantly contribute to the antitumour effect of cisplatin and hyperthermia, but it partially contributed to the immunomodulatory effect and to the reduction of cisplatin toxicity and to a slight increase in animal survival. This treatment schedule did not affect microvessel density, but it inhibited tumour growth and modulated macrophage polarization to the M1 phenotype. Furthermore, it abolished the resistance of tumour cells to cisplatin by modulating HDAC activity and the concentration of HSP70 and HSP90 chaperones, contributing to the increased lifespan of mice. However, the precise mechanism of the interaction between resveratrol, cisplatin, and hyperthermia needs to be investigated further.

Selenium and vitamin E ameliorate lead acetate-induced hepatotoxicity in rats via suppression of oxidative stress, mRNA of heat shock proteins, and NF-kB production

BackgroundLead exposure results in a terrible rise in heat shock protein levels. ObjectiveThis research was conducted to look at the effects of lead poisoning on heat shock response, oxidative stress, and inflammatory markers in albino rats, as well as the power of selenium and vitamin E to resist lead toxic effects. MethodsEight groups of albino rats are used. Each group contained six rats where the first group represented the negative control, and the other groups were treated with olive oil, vitamin E, selenium, lead, (vitamin E + lead), (selenium + lead), and (vitamin E + selenium + lead). All the treatments lasted for 28 days. Then, the mRNA expression of interested heat shock proteins (HSP90, HSP70, and HSP60) was assessed. For oxidative stress disruption, we investigated nitric oxide (NO) and malondialdehyde (MDA) content, and enzymatic and non-enzymatic antioxidants activity respectively in rat livers. Resultsour results revealed the synergetic protective effect of the combination of two antioxidants (vitamin E and selenium) against lead poising. This was clear in regulating HSPs expression, inflammatory markers, glucose, lipid profile, liver functions, and antioxidant enzymes more than the treatment with one antioxidant. ConclusionPb is a toxic material that can induce HSPs and inflammatory markers expression. Selenium and vitamin E can give excellent effects in ameliorating Pb toxicity when used together.

Preparation of Phage Antibodies to Heat Shock Proteins and Studying the Dynamics of their Accumulation in Mice with Xenotransplant Tumors

Using a naïve human scFv phage library, affinity selection of miniantibodies specific to heat shock proteins isolated from MH22a hepatoma cells and Sp2/0-Ag14 plasmacytoma cells was carried out. Using the obtained phage antibodies by dot-immunoassay and enzyme-linked immunosorbent assay, the dynamics of the concentration of heat shock proteins in the blood serum of mice with implanted tumor cells of the MH22a line was studied. Starting from the 14th day after xenotransplantation, there was a gradual increase in the level of heat shock proteins in the blood serum. It was found that after implantation of tumor cells, tumor growth was accompanied by a significant increase in the accumulation of heat shock proteins in the blood serum. It has been shown that miniantibodies specific to heat shock proteins are an effective tool for determining and monitoring the accumulation of heat shock proteins in the blood serum of animals.

Oxidative Stress and Cellular Protein Accumulation Are Present in Keratoconus, Macular Corneal Dystrophy, and Fuchs Endothelial Corneal Dystrophy

The aim of the study was to investigate oxidative stress as well as cellular protein accumulation in corneal diseases including keratoconus (KC), macular corneal dystrophy (MCD), and Fuchs endothelial corneal dystrophy (FECD) at their primary affecting sites. Corneal buttons from KC, MCD, and FECD patients, as well as healthy controls, were analyzed immunohistochemically to evaluate the presence of oxidative stress and the function of the proteostasis network. 4-Fydroxynonenal (4-HNE) was used as a marker of oxidative stress, whereas the levels of catalase and heat-shock protein 70 (HSP70) were analyzed to evaluate the response of the antioxidant defense system and molecular chaperones, respectively. Sequestosome 1 (SQSTM1) levels were determined to assess protein aggregation and the functionality of autophagic degradation. Basal epithelial cells of the KC samples showed increased levels of oxidative stress marker 4-HNE and antioxidant enzyme catalase together with elevated levels of HSP70 and accumulation of SQSTM1. Corneal stromal cells and endothelial cells from MCD and FECD samples, respectively, showed similarly increased levels of these markers. All corneal diseases showed the presence of oxidative stress and activation of the molecular chaperone response to sustain protein homeostasis. However, the accumulation of protein aggregates suggests insufficient function of the protective mechanisms to limit the oxidative damage and removal of protein aggregates via autophagy. These results suggest that oxidative stress has a role in KC, MCD, and FECD at the cellular level as a secondary outcome. Thus, antioxidant- and autophagy-targeted therapies could be included as supporting care when treating KC or corneal dystrophies.

Comparison of behavioural and transcriptional responses to a heat stressor between freshly collected and an inbred strain of Lymnaea

Different populations of organisms occurring across varying thermal regimes show diversity in responses to heat stress. We use a “common garden experimental” approach designed to deal with phenotypic plasticity to study in Lymnaea stagnalis (Linnaeus, 1758) the behavioural and molecular responses to a heat shock in laboratory-inbred snails (W-strain) and freshly collected snails (Stony strain) from ponds. In the W-strain, which has been reared under standardized temperatures for generations, the exposure to 30 °C for 1 h (heat shock, HS) when experienced after a novel “taste” results in a taste-specific aversion known as the “Garcia effect”. This learned avoidance requires the upregulation of heat shock proteins (HSPs). In contrast, freshly collected Stony strain, which experiences temperature fluctuations regularly, does not exhibit a Garcia effect. Here, we found that (1) Stony-strain snails have higher basal mRNA levels of HSPs than W-strain ones; (2) in the W-strain, the training procedure to cause the Garcia effect upregulates the mRNA levels of HSPs and key neuroplasticity-related genes such as CREB1 and GRIN1; (3) in Stony-strain snails, the same training procedure fails to alter the mRNA levels of those targets. These data suggest that Stony-strain snails do not perceive the HS as a stressor because of the higher HSP basal mRNA levels, which may confer a higher thermal tolerance.

Examination of two different proteasome inhibitors in reactivating mutant human cystathionine β-synthase in mice.

Classic homocystinuria is an inborn error of metabolism caused mainly by missense mutations leading to misfolded and/or unstable human cystathionine β-synthase (CBS) protein, causing the accumulation of excess total homocysteine (tHcy) in tissues. Previously, it has been shown that certain missense containing human CBS proteins can be functionally rescued in mouse models of CBS deficiency by treatment with proteasome inhibitors. The rescue by proteasome inhibitors is thought to work both by inhibiting the degradation of misfolded CBS protein and by inducing the levels of heat-shock chaperone proteins in the liver. Here we examine the effectiveness of two FDA approved protease inhibitors, carfilzomib and bortezomib, on various transgenic mouse models of human CBS deficiency. Our results show that although both drugs are effective in inducing the liver chaperone proteins Hsp70 and Hsp27, and are effective in inhibiting proteasome function, bortezomib was somewhat more robust in restoring the mutant CBS function. Moreover, there was no significant correlation between proteasome inhibition and CBS activity, suggesting that some of bortezomib's effects are via other mechanisms. We also test the use of low-doses of bortezomib and carfilzomib on various mouse models for extended periods of time and find that while low-doses are less toxic, they are also less effective at restoring CBS function. Overall, these results show that while restoration of mutant CBS function is possible with proteasome inhibitors, the exact mechanism is complicated and it will likely be too toxic for long-term patient treatment.