Hsp70 Expression Research Articles

Salidroside regulates tumor microenvironment of non-small cell lung cancer via Hsp70/Stub1/Foxp3 pathway in Tregs

BackgroundThe treatment of non-small cell lung cancer (NSCLC) is challenging due to immune tolerance and evasion. Salidroside (SAL) is an extract in traditional Chinese medicine and has a potential antitumor effect. However, the mechanism of SAL in regulating the immunological microenvironment of NSCLC is yet to be clarified.MethodsThe mouse model with Lewis lung cancer cell line (3LL) in C57BL/6 mice was established. And then, the percentage of tumor-infiltrating T cell subsets including Treg was detected in tumor-bearing mice with or without SAL treatment. In vitro, the effect of SAL on the expression of IL-10, Foxp3 and Stub1 and the function of Treg were detected by flow cytometry. Network pharmacology prediction and molecular docking software were used to predict the target of SAL and intermolecular interaction. Furthermore, the effect of SAL on the expression of Hsp70 and the co-localization of Stub1-Foxp3 in Treg was confirmed by flow cytometry and confocal laser microscopy. Finally, Hsp70 inhibitor was used to verify the above molecular expression.ResultsWe discovered that SAL treatment inhibits the growth of tumor cells by decreasing the percentage of tumor-infiltrated CD4+Foxp3+T cells. SAL treatment downregulates the expression of Foxp3 in Tregs, but increases the expression of Stub1, an E3 ubiquitination ligase upstream of Foxp3, and the expression of Hsp70. Inhibiting the expression of Hsp70 reverses the inhibition of SAL on Foxp3 and disrupts the colocalization of Stub1 and Foxp3 in the nucleus of Tregs.ConclusionsSAL inhibits tumor growth by regulating the Hsp70/stub1/Foxp3 pathway in Treg to suppress the function of Treg. It is a new mechanism of SAL for antitumor therapy.

The first reporting of prevalence Vibrio species and expression of HSP genes in rayed pearl oyster (Pinctada radiata) under thermal conditions.

The main goal of the present study was to evaluate the influence of thermal exposure on Vibrio population and HSP genes expression (HSP 90, HSP70, and HSP20) in rayed pearl oyster (P. radiata). To this end, the oysters were reared for 30 days at temperatures of 22°C (control), 25°C, 27°C, and 29°C. The results showed that five dominate Vibrio strains including Vibrio hepatarius, V. harveyi, V. alginolyticus, V. parahaemolyticus, and V. rotiferianus were identified. The highest population of V. parahaemolyticus, V. alginolyticus, and V. harveyi, was found in 29οC group. According to real-time PCR, mantle exhibited the highest expression levels of HSP20, HSP70, and HSP90 genes. A higher level of HSP20 expression was observed at high temperatures (25°C, 27°C, and 29°C) in the gonad and mantle compared to the control group (22°C) while decrease in HSP90 expression level was recorded in 25°C, 27°C, and 29°C groups. HSP20 expression level in adductor muscle was remarkably down-regulated in 27°C and 29°C groups. In this tissue, HSP70 was detected at highest levels in the 29οC group. In mantle, HSP90 gene expression was lowest at 22°C water temperature. Several Vibrio strains have been identified from pearl Gulf oyster that haven't been previously reported. The identification of dominant Vibrio species is essential for epidemiological management strategies to control and prevent Vibrio outbreaks in pearl oyster farms. The expression pattern of HSP genes differs in rayed pearl oyster tissues due to differences in their thermal tolerance capability and physiological and biological characteristics. The present study provides useful molecular information for the ecological adaptation of rayed pearl oysters after exposure to different temperature levels.

Effects of Intermittent Temperature and Humidity Regulation on Tomato Gray Mold.

Temperature and humidity play an important role in plant-pathogen interactions. However, regulating the temperature and humidity specifically to inhibit the development of plant diseases remains unclear. In this study, we explored the influence of intermittent temperature and humidity variation on tomato gray mold. Intermittent regulation of temperature and humidity (increasing temperature with decreasing humidity for different periods within 24 h) inhibited the disease severity of plants and the infection process of Botrytis cinerea. The 4-h treatment (increasing temperature accompanied by decreasing humidity for 4 h and recovering for 4 h, and so on) effectively inhibited the development of tomato gray mold, reduced the biomass of B. cinerea, delayed the differentiation time of mycelia, and inhibited the accumulation of hydrogen peroxide in tomato leaves at the later stage of infection. The increased expressions of heat-shock protein (HSP) genes HSP20, HSP70, HSP90, BAG6, and BAG7 in tomato were mainly caused by environmental changes and environment-plant-pathogen interactions, and the increased expression of the latter was greater than that of the former in the 2-h (increasing temperature accompanied by decreasing humidity for 2 h and recovering for 2 h, and so on) and 4-h treatments. Pathogen infection induced the expression of defense-related genes in tomato, and the increase in the expressions of FLS2, FEI1, PI2, Pti5, and WRKY75 induced by B. cinerea in the 4-h treatment was greater than that under unregulated temperature and humidity conditions. In general, intermittent temperature and humidity variation can effectively inhibit the development of tomato gray mold, and the 4-h treatment had the best inhibitory effect.

HSP90 Antisense Oligonucleotide Enhances the Inhibitory Effect of Photothermal Therapy on Triple-Negative Breast Cancer Cells

Photothermal therapy is a potential strategy to treat triple-negative breast cancer. The expression of HSP90 in the tumor cells has been identified as a culprit for reducing the effectiveness of photothermal therapy, so using reagents with photothermal conversion to deliver HSP inhibitors can achieve good tumor suppression. However, the potential toxicity of small molecule HSPs inhibitors limits their further application development. Therefore, this study constructed a nanoplatform by graphene oxide (GO) to deliver antisense oligonucleotide (ASO) of HSP90 to improve the therapeutic effect. The photothermal performance of GO was confirmed by infrared thermal imager. MTT results showed that tumor cell viability was lower after the addition of ASO under irradiation. In addition, the result of western blot revealed that the expression of HSP90 decreased by 32% compared to the blank group under the action of ASO. All the above results indicated that our strategy enhances the inhibition of triple-negative breast cancer cells via suppressing the expression of HSP90 in photothermal therapy.

Bisphenol-A exposure alters liver, kidney, and pancreatic Klotho expression by HSP60-activated mTOR/autophagy pathway in male albino rats.

The effect of bisphenol-A (BPA) on Klotho protein (aging-suppressing protein) expression in different body organs has not been sufficiently addressed by literature studies. The study investigated the impact of BPA on Klotho expression in multiple organs including the liver, kidney, and pancreas and suggested the involved molecular pathways. Twenty-seven male Wistar albino rats were divided into 3 equal groups: control, low-dose BPA (4.5 µg/L), and high-dose BPA (8 µg/L) groups in drinking water for 45 consecutive days. Liver, kidney, and pancreatic specimens were prepared for a gene study of Klotho, HSP60, mTOR, and ULK1 mRNA expressions. Also, the tissue specimens were measured for malondialdehyde (MDA), superoxide dismutase (SOD), and nitric oxide (NO) levels. Paraffin-embedded sections were also prepared and subjected to Hematoxylin and Eosin (H&E) staining and immunohistochemical detection of Klotho and HSP60. The results revealed an alteration in the MDA, SOD, NO tissue levels, disturbed gene expression profile, and apoptotic changes in the histological findings of the examined organs which were obvious (p < 0.05) in the high-dose group. The anti-aging Klotho gene/protein expression was reduced (p < 0.05) more in the high-dose BPA group than in the low dose. In contrast, HSP60 gene/protein expression was significantly increased (p < 0.05) more in the high dose. It was concluded that BPA exposure contributed to cell stress and markedly reduced Klotho protein expression in liver, kidney, and pancreatic tissues, possibly by modulation of the HSP60-activated mTOR/autophagy signaling.

The Assessment of Metal Resistance through the Expression of Hsp-70 and HO-1 Proteins in Giant Reed

None of our investigations have identified stress in response to the HSP70 and HO-1 proteins in metals under stress in our study, which aimed to understand the genetic basis of the metal tolerance of Arundo donax. Thus, the present work aimed to investigate the levels of expression of two important stress-related proteins, HO-1 and HSP70, in A. donax after exposure to various metals. The plants were collected from uncontaminated sites in Abbottabad, Pakistan. Their rhizomes were grown in Hoagland solution, and upon attaining suitable biomass, the plants were used to investigate the effects of metals on protein expression. The metal treatments were carried out with synthetic wastewater containing four Cr treatments (0, 34, 66, 134, and 267 mgL−1), namely, Cd, As, Pb, Cu and Ni (0, 25, 50, 75, and 100 mgL−1), and the plants were grown for three weeks. The treatments were applied according to a randomized block design (RBD) based on hydroponics. The selected protein expression was examined after 10 days of metal exposure. For the HSP70 and HO-1 protein studies, leaves were separated following a previously reported standard procedure. The maximum level of HO-1 and HSP70 expressions was noted at 66 mgL−1 of Cr, and then it slightly declined. Significantly, high protein expression was observed at Cd exposure concentrations of 50 to 100 mgL−1. For Cu, As and Ni, significantly high HO-1 and HSP70 expressions were noted at metal exposure concentrations of 75 to 100 mgL−1. The expression levels of these two stress-related proteins showed a linear increase with increasing metal exposure in the giant reed. It is clear from the present research that HSP70 and HO-1 proteins may contribute significantly to plant tolerance to metal stress, in addition to other possible tolerance mechanisms.

RGA1 Negatively Regulates Thermo-tolerance by Affecting Carbohydrate Metabolism and the Energy Supply in Rice

BackgroundSignal transduction mediated by heterotrimeric G proteins, which comprise the α, β, and γ subunits, is one of the most important signaling pathways in rice plants. RGA1, which encodes the Gα subunit of the G protein, plays an important role in the response to various types of abiotic stress, including salt, drought, and cold stress. However, the role of RGA1 in the response to heat stress remains unclear.ResultsThe heat-resistant mutant ett1 (enhanced thermo-tolerance 1) with a new allele of the RGA1 gene was derived from an ethane methyl sulfonate-induced Zhonghua11 mutant. After 45 °C heat stress treatment for 36 h and recovery for 7 d, the survival rate of the ett1 mutants was significantly higher than that of wild-type (WT) plants. The malondialdehyde content was lower, and the maximum fluorescence quantum yield of photosystem II, peroxidase activity, and hsp expression were higher in ett1 mutants than in WT plants after 12 h of exposure to 45 °C. The RNA-sequencing results revealed that the expression of genes involved in the metabolism of carbohydrate, nicotinamide adenine dinucleotide, and energy was up-regulated in ett1 under heat stress. The carbohydrate content and the relative expression of genes involved in sucrose metabolism indicated that carbohydrate metabolism was accelerated in ett1 under heat stress. Energy parameters, including the adenosine triphosphate (ATP) content and the energy charge, were significantly higher in the ett1 mutants than in WT plants under heat stress. Importantly, exogenous glucose can alleviate the damages on rice seedling plants caused by heat stress.ConclusionRGA1 negatively regulates the thermo-tolerance in rice seedling plants through affecting carbohydrate and energy metabolism.

Stocking density affects immune and stress-related gene expression of Butter catfish (Ompok bimaculatus) fry in biofloc landscapes

Scientific research into fish wellness is critical, and the concerns about crowding-related stress due to increased stocking density are inevitable. Taking this into consideration, the study defines the physiological signature of Ompok bimaculatus (Butter catfish) in a biofloc system when subjected to varying levels of stocking density. Fish (mean weight = 1.21 g ± 0.08, n = 600) were randomly stocked in 40-L glass aquaria at stocking densities of 0.5 g/L (T1), 1 g/L (T2), 1.5 g/L (T3), and 2 g/L (T4) and fed a 35% protein diet. After the 90-day trial, the physio-biochemical, molecular, and tissue-level changes were assessed. An integrated biomarker response (IBR) analysis for the key stress indicators aided us in better understanding them. There was a significant difference in blood count between T1 and T4 (total erythrocyte count, hemoglobin, and packed cell volume). T1 had higher levels of globulin and total plasma protein, but T2 had higher levels of albumin. Only in T1 did the respiratory burst and lysozyme activity appear to be higher (p < 0.05). Increased stocking densities had a significant impact on the liver function enzymes, GOT and GPT (p < 0.05). In comparison to lower densities (T1 & T2), higher stocking density (T3 & T4) was found to raise glucose and cortisol levels (p < 0.05). Antioxidant enzymes such as catalase, glutathione-S-transferase, and malondialdehyde were found to be more pronounced in lower density tissues (T1). Furthermore, the IBR plots show that lower densities have better health than higher densities. At higher stocking densities, mRNA expression of HSP70, IL-1, and IL-20 increased (p < 0.05) in kidney and liver tissues. The Nrf-2 and Tlr-9 genes were also upregulated. Also, when stocking density was increased, tissue-level histo-architectural changes were more pronounced than when stocking density was kept low. The findings of this study show that the welfare of Butter catfish cultured at high density in biofloc systems suffers from severe stress, and therefore draw more attention to the development of a species-specific standard rearing methodology in the pursuit of a profitable aqua-farming enterprise.

LncRNA MALAT1 Promoted Neuronal Necroptosis in Cerebral Ischemia-reperfusion Mice by Stabilizing HSP90.

The objective of this research was to investigate the role of lncRNA MALAT1 and HSP90 in the regulation of neuronal necroptosis in mice with cerebral ischemia-reperfusion (CIR). We used male C57BL/6J mice to establish a middle cerebral artery occlusion (MCAO) model and conducted in vitro experiments using the HT-22 mouse hippocampal neuron cell line. The cellular localization of NeuN and MLKL, as well as the expression levels of neuronal necroptosis factors, MALAT1, and HSP90 were analyzed. Cell viability and necroptosis were assessed, and we also investigated the relationship between MALAT1 and HSP90. The results showed that MALAT1 expression increased after MCAO and oxygen-glucose deprivation/re-oxygenation (OGD/R) treatment in both cerebral tissues and cells compared with the control group. The levels of neuronal necroptosis factors and the co-localization of NeuN and MLKL were also increased in MCAO mice compared with the Sham group. MALAT1 was found to interact with HSP90, and inhibition of HSP90 expression led to decreased phosphorylation levels of neuronal necroptosis factors. Inhibition of MALAT1 expression resulted in decreased co-localization levels of NeuN and MLKL, decreased phosphorylation levels of neuronal necroptosis factors, and reduced necroptosis rate in cerebral tissues. Furthermore, inhibiting MALAT1 expression also led to a shorter half-life of HSP90, increased ubiquitination level, and decreased phosphorylation levels of neuronal necroptosis factors in cells. In conclusion, this study demonstrated that lncRNA MALAT1 promotes neuronal necroptosis in CIR mice by stabilizing HSP90.

Inhibition of the MLCK/MLC2 pathway protects against intestinal heat stroke-induced injury in rats

Intestinal barrier dysfunction often exists in the heat stroke (HS) pathological process, which increases intestinal permeability and induces endotoxemia. The upregulation of MLCK is a crucial player affecting intestinal permeability. This study aimed to explore whether inhibiting myosin light chain kinase (MLCK) can improve HS-induced intestinal injury in rats. Twelve-week-old Wistar male rats were divided into three groups: the control group, the HS model group, and the treatment group [HS model + ML-7 (MLCK inhibitor)]. HS impaired the tight junctions in the rat gut and increased permeability. Additionally, increased inflammatory factors in serum, activation of apoptosis, and downregulation of tight junction proteins were observed in intestinal cells. ML-7 significantly inhibited the MLCK/p-MLC2 signaling pathway, increased the expression of tight junction proteins, reduced intestinal permeability, reduced apoptosis and alleviated the intestinal damage caused by HS. ML-7 inhibited HS-induced apoptosis of intestinal epithelial cells by regulating the ERK/p38/HSP70 axis. Furthermore, inhibition of MLCK upregulated HSP70 expression through activation of the ERK pathway and inhibited cell apoptosis by abolishing the p38 MAPK pathway. In conclusion, inhibiting the MLCK/p-MLC2 signaling pathway reduces HS-induced intestinal permeability and protects the intestinal mucosal barrier.

Tanespimycin and rapamycin exhibit antifungal activity against Colletotrichum gloeosporioides and enhance mango resistance to anthracnose via differentially modulating heat shock response

Postharvest mango (Mangifera indica L.) fruit suffer tremendous loss due to anthracnose inflicted by Colletotrichum spp. Fungicides and heat treatment are often used to control the disease, while biocontrol agents are eco-friendly alternatives. Streptomyces are antagonistic organisms against plant pathogens; however, the mechanisms and active components are unclear. Here, we evaluated the antifungal actions of S. hygroscopicus metabolites tanespimycin and rapamycin against anthracnose. Within the range of 0.01–0.1 μM, tanespimycin and rapamycin reduced fungal growth by at least 60.4% and 40.4%, respectively in vitro. In mango fruit, tanespimycin upregulated MiHSP101, MiHSP90, MiHSP70, and MiHSP18.2 by 5.8-, 1.9-, 4.8-, and 3.3-fold, respectively, and increased HSP101 and HSP70 protein by 2.2-fold. On the other hand, rapamycin did not affect HSP expression. Pathogen-challenge assays revealed tanespimycin and rapamycin exerted antifungal and defense-eliciting dual actions, where pre-challenge treatment of tanespimycin and rapamycin inhibited pathogen growth by 73.7% and 78.4%, respectively in mango 5 d post infection. Finally, in a 20 °C storage experiment, tanespimycin and rapamycin extended the fruit shelf life by 6 d and exerted comparable antifungal effects as hot water treatment without deteriorating fruit quality. In conclusion, we elucidated the antifungal and defense-eliciting actions of S. hygroscopicus metabolites and revealed mechanistic discrepancies regarding HSP regulation. These findings support the development of new disease management strategies and advance the knowledge regarding the roles of HSP in crop defense.

Protective effects of epigallocatechin gallate in the mice induced by chronic scrotal hyperthermia

One of the most common complications of chronic scrotal hyperthermia (SHT) is a serious disorder in the male reproductive system. The most important factor in the occurrence of these disorders is oxidative stress. Currently, we investigated the effects of epigallocatechin gallate (EGCG), as a highly potent antioxidant, against cells and tissue disorders in mice affected by chronic SHT. Fifty-six male adult NMRI mice were allocated into seven equal groups. Except the non-treated (Control) group, six other groups were exposed to heat stress. Two treated groups including Preventive and Curative received oral administration of EGCG (50 mg/kg/day) starting immediately before heat exposure and fifteen consecutive days after the end of the heat exposure, respectively. For each treated group, two subgroups including positive control (Pre/Cur + PC groups) and vehicle (Pre/Cur + vehicle groups) were considered. At the end of the study, sperm characteristics, testosterone levels, stereological parameters, apoptosis, oxidant state, and molecular assessments were performed. We found that the sperm parameters, testosterone levels, the numerical density of spermatogonia, primary spermatocytes, spermatids, sertoli, leydig cells, and seminiferous tubules, biochemical factors (except MDA), and expression of c-kit gene were significantly higher in the Preventive and Curative groups, especially in Preventive ones, compared to other groups (P < 0.05). This is while expression of HSP72 and NF-κβ genes, MDA levels, as well as density of apoptotic cells considerably decreased in both EGCG-treated groups compared to other groups and it was more pronounced in Preventive ones (P < 0.05). Generally, EGCG attenuated cellular and molecular disorders induced by heat stress in the testis and it was more pronounced in Preventive status.

Expression of Hsp70 in the red swamp crayfish, Procambarus clarkii in response to immune challenge

A heat shock protein 70 (Hsp70) was identified from Procambarus clarkii (named as PcHsp70). The open reading frame of the PcHsp70 cDNA was 1917 bp, encoding a 638 aa protein with signature sequences of the Hsp70 family. Tissue distribution results showed that PcHsp70 has relatively high expression in immune-related tissues such as hepatopancreas, gill, and hemocytes. The expressions of PcHsp70 in hepatopancreas and gill were significantly induced by Gram-positive bacterium, Gram-negative bacterium, and white spot syndrome virus stimulation, indicating that PcHsp70 may be involved in the immune response to pathogen invasion. The research will provide a foundation for understanding the resistance and stress mechanism of Hsp70 in P. clarkii.

Thymoquinone attenuates doxorubicin-induced lung damage via heat shock proteins, inflammation and endoplasmic reticulum stress

The aim of this study is to determine possible therapeutic effects of THQ on doxorubicin induced lung damage in rats. Rats were divided into five groups (n = 8): control, THQ, olive oil, DOX (a single dose of 15 mg/kg intraperitoneally (i.p.) on seventh day of the experiment), and DOX+THQ (10 mg/kg THQ per day and 15 mg/kg DOX i.p. on seventh day). Animals were euthanized, and lung tissues were evaluated histopathologically. Caspase3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2 and TNFα immunostaining were performed to determine the expression levels of these proteins among groups. TUNEL method was used for evaluation of apoptotic index. Moreover, total antioxidant status (TAS), total oxidant status (TOS) and IL6 in lung tissue were measured by ELISA assay. The DOX group had histopathological deterioration compared to the control group. There was an increase in apoptotic index, caspase 3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2 and TNFα expressions in the DOX group. While TAS level of the DOX group decreased, IL6 and TOS level increased when compared with the other groups. However, there was improvement in lung tissue in DOX + THQ group compared to the DOX group. There was a decrease in apoptotic index, caspase 3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2, TNFα expressions and TOS, IL6 in DOX+THQ group compared to the DOX group. We suggest that THQ can be used as a protective agent to reduce the toxic effects of DOX.

Dietary protein, lipid and insect meal on growth, plasma biochemistry and hepatic immune expression of lake whitefish (Coregonus clupeaformis)

Studies are lacking that investigate the dietary nutrient requirements of lake whitefish (Coregonus clupeaformis), a newly farmed fish species in Ontario, Canada. Dietary levels of protein and lipid must be optimized to ensure high growth performance for the commercial success of this species. Additionally, the inclusion of insect meal in the diet may improve growth and immune response. The objective of this study was to evaluate the effects of dietary protein:lipid ratios and insect meal as a feed additive on the growth performance and hepatic immune function of juvenile lake whitefish (301 ± 10 g). A 16-week (112 day) trial was performed with five diets including a commercial control diet (BCC), and four experimental diets with high or low levels of protein (54 and 48%, respectively) and lipid (18 and 12%, respectively). The high protein dietary groups contained 5% of full-fat black soldier fly larvae (Hermetia illucens). Fish weights, viscera, liver, and blood were collected for further analysis. Specific growth rate, thermal growth coefficient and weight gain were significantly higher in fish fed with the BCC and high protein high lipid (HPHL) diets. However, viscerosomatic index was found to be significantly higher in fish fed the BCC diet, thus HPHL is more optimal for non-visceral weight gain. Higher levels of plasma phosphorus, aspartate aminotransferase and potassium indicated poor growth and stress in fish fed low lipid diets. Relative expression of HSP70, involved in cellular repair, was significantly downregulated in fish fed high lipid diets, and no effects were found on the expression of innate immune and oxidative stress genes. Also, IL8 (CXCL8) and catalase were upregulated (non-significant) in fish fed the HPHL diet with the largest weight gain. No effects of insects were found on growth, plasma biochemistry or gene expression, which suggests 5% dietary inclusion was too low. Overall, we recommend a HPHL diet for the cultivation of lake whitefish based on improved growth performance, low viscera weight, improved plasma biochemistry and downregulation of cellular repair genes.

Anti-heat shock protein 10 IgG in chronic spontaneous urticaria: Relation with miRNA-101-5p and platelet-activating factor.

Anti-heat shock protein (HSP) autoantibodies are detected in autoimmune diseases. We sought to ascertain whether anti-HSP10 IgG is present in patients with CSU and to elucidate the role of HSP10 in CSU pathogenesis. Using a human proteome microarray, six potential autoantibodies had higher expression in 10 CSU samples compared with 10 normal controls (NCs). Among them, HSP10 IgG autoantibody was quantified by immune dot-blot assay in sera from 86 CSU patients and 44 NCs. The serum levels of HSP10 and microRNA-101-5p were measured in CSU patients and NCs. The effects of HSP10 and miR-101-5p on mast cell degranulation in response to IgE, compound 48/80, and platelet-activating factor (PAF) were investigated. CSU patients had higher IgG positivity to HSP10 (40.7% vs. 11.4%, p = .001), lower serum HSP10 levels (5.8 ± 3.6 vs. 12.2 ± 6.6 pg/mL, p < .001) than in NCs, and their urticaria severity was associated with anti-HSP10 IgG positivity, while HSP10 levels were related to urticaria control status. MiR-101-5p was increased in CSU patients. PAF enhanced IL4 production in PBMCs from CSU patients. IL-4 upregulated miR-101-5p and reduced HSP10 expression in keratinocytes. Transfection of miR-101-5p reduced HSP10 expression in keratinocytes. MiR-101-5p promoted PAF-induced mast cell degranulation, while HSP10 specifically prevented it. A new autoantibody, anti-HSP10 IgG was detected in CSU patients, which showed a significant correlation with UAS7 scores. A decreased serum HSP10 level was associated with upregulation of miR-101-5p due to increased IL-4 and PAF in CSU patients. Modulation of miR-101-5p and HSP10 may be a novel therapeutic approach for CSU.

Prediction and verification of the active ingredients and potential targets of Erhuang Quzhi Granules on non-alcoholic fatty liver disease based on network pharmacology.

Erhuang Quzhi Granules (EQG) is a compound composed of 13 traditional Chinese medicines developed by the First Affiliated Hospital of Shihezi University. In clinical practice, EQG has been applied to the treatment of hyperlipidemia and non-alcoholic fatty liver disease (NAFLD), and could significantly improve the serum biochemical indicators of NAFLD patients. This study aims to explore the bioactive compounds, potential targets, and molecular mechanisms of EQG against NAFLD through network pharmacology, molecular docking, and experimental verification. The chemical components of EQG came from the literature and quality standard. Bioactive compounds were screened based on the absorption, distribution, metabolism, and excretion (ADME) feature, and their potential targets were predicted using the substructure-drug-target network-based inference (SDTNBI). The core targets and signaling pathways were obtained through the analysis of protein-protein interaction (PPI), gene ontology (GO) function, and Kyoto encyclopedia of genes and genomes (KEGG) pathway. The results were further confirmed by literature retrieval, molecular docking, and in vivo experiments. The results of network pharmacology showed 12 active ingredients and 10 core targets for EQG in treating NAFLD. And EQG mainly regulates lipid and atherosclerosis-related pathways to improve NAFLD. The collected literature verified the regulatory effect of the active components of EQG on core targets TP53, PPARG, EGFR, HIF1A, PPARA, and MTOR. Molecular docking results showed that Aloe-Emodin (AE), Emodin, Physcion, and Rhein (RH) had stable binding structures with the core targets HSP90AA1. In vivo experiment showed that AE and RH reduced aspartate transaminase (AST), alanine aminotransferase (ALT), interleukin (IL)-1β, IL-6, IL18, and tumor necrosis factor α (TNF-α) in the serum or liver of NAFLD mice, improved liver lipid deposition and fibrosis, and inhibit gene expression of nuclear factor kappa B (NF-κB), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), IL-1β, TNF-α and protein expression of HSP90, NF-κB and Cleaved caspase-1. This study comprehensively revealed the biological compounds, potential targets, and molecular mechanisms of EQG in the treatment of NAFLD, providing a reference basis for the promotion of EQG in the clinic.

A redox-responsive self-assembling COA-4-arm PEG prodrug nanosystem for dual drug delivery suppresses cancer metastasis and drug resistance by downregulating hsp90 expression.

Metastasis and resistance are main causes to affect the outcome of the current anticancer therapies. Heat shock protein 90 (Hsp90) as an ATP-dependent molecular chaperone takes important role in the tumor metastasis and resistance. Targeting Hsp90 and downregulating its expression show promising in inhibiting tumor metastasis and resistance. In this study, a redox-responsive dual-drug nanocarrier was constructed for the effective delivery of a commonly used chemotherapeutic drug PTX, and a COA-modified 4-arm PEG polymer (4PSC) was synthesized. COA, an active component in oleanolic acid that exerts strong antitumor activity by downregulating Hsp90 expression, was used as a structural and functional element to endow 4PSC with redox responsiveness and Hsp90 inhibitory activity. Our results showed that 4PSC/PTX nanomicelles efficiently delivered PTX and COA to tumor locations without inducing systemic toxicity. By blocking the Hsp90 signaling pathway, 4PSC significantly enhanced the antitumor effect of PTX, inhibiting tumor proliferation and invasiveness as well as chemotherapy-induced resistance invitro. Remarkable results were further confirmed invivo with two preclinical tumor models. These findings demonstrate that the COA-modified 4PSC drug delivery nanosystem provides a potential platform for enhancing the efficacy of chemotherapies.

The Effect of Sericin on Heat Shock Proteins 60 and 90, and TLR2 and TLR4 Induced by Chronic Heat Stress in Male Mice

Background: Heat stress followed by heat stroke, which is caused by an increase in body temperature to more than 40 °C, is a medical emergency that affects many people in work situations and weather conditions and results in irreversible neurological and cognitive damage. Methods: In general, 75 male albino mice in the weight range of 28-30 g were used in the present study. The mice were divided into 5 groups of 15, including the control group, heat stress (HS) group, HS group+sericin 100, HS group+sericin 150, and HS group+sericin 200. Toll-like receptor 2 (TLR2) and Toll-like receptor (TLR4) expression levels were measured using the real-time polymerase chain reaction (RT-PCR), and the expression of hsp60 and hsp90 proteins was measured using a western blot. Results: The expression level of TLR2 and TLR4 receptors increased significantly with the passage of time as a result of stimulation. Western blot results showed that the expression levels of HSP60 and HSP90 increased as well. Conclusion: The results of RT-PCR revealed that HS significantly increases the expression of innate immune system receptors such as TLR2 and TLR4.

Metabolic and transcriptomic response of two juvenile anadromous brook charr (Salvelinus fontinalis) genetic lines towards a chronic thermal stress

Many salmonid species are particularly susceptible to chronic and acute temperature changes caused by global warming. We aimed to study the differences in metabolic and transcriptomic responses of a chronic heat stress on a control and selected (absence of early sexual maturation and growth) line of brook charr Salvelinus fontinalis (Mitchill, 1814). We exposed individuals to different temperatures for 35 days (15, 17, and 19 °C). High temperature reduced the growth rate (in length) and the Fulton condition factor. Both maximal metabolic rate and the aerobic scope were higher in fish reared at 17 °C, while they decreased in fish maintained at 19 °C. The relative gene expression of cytochrome c oxidase was lower at 19 °C than at 15 °C. The relative gene expressions of both liver and gill hsp90 was higher at the highest temperature. The standard metabolic rate, while not affected by temperature, was higher for the control line over the selected line. Only in the control line, the relative expression of catalase and of receptor of insulin-like growth factor-1 increased at 19 °C. Our results showed that the selected line was able to cope more effectively with the oxidative stress caused by the rise in temperature.