Relationship between hyperuricemia, HSP70 and NLRP3 inflammasome in arterial hypertension / La relación hiperuricemia-HSP70-NLRP3 inflamosoma en la hipertensión arterial

Abstract Arterial hypertension is the most important cardiovascular risk factor in chronic non-communicable diseases and is estimated to be responsible for 10.4 million deaths annually. The global prevalence of hypertension is 30% and the majority of people with hypertension do not have a clear identifiable cause and are considered to have primary hypertension. Experimental and clinical investigations from several research groups, including ours, have established that inflammation and autoimmune reactivity play a role in the sodium retention and hemodynamic responses that drive primary hypertension. Hyperuricemia and heat stress proteins (HSP), particularly HSP70, are both associated with the activation of innate immunity that plays a role in the development of inflammatory reactivity in the hypertensive patient. Clinical studies have shown an association between the expression of HSP70 and anti-HSP70 antibodies and primary hypertension. This brief review aims to examine the interrelation between hyperuricemia and extracellular overexpression of HSP70 in the activation of the inflammasome that may have a central role in the pathophysiology of primary hypertension.

Resumen La hipertensión arterial es el factor de riesgo cardiovascular más importante de las enfermedades crónicas no transmisibles y se estima que es responsable de 10.4 millones de muertes al año. La prevalencia mundial de la hipertensión es del 30%; la mayoría de las personas con hipertensión no tienen una causa claramente identificable y se considera que tienen hipertensión primaria. Las investigaciones experimentales y clínicas de varios grupos de investigación, incluido el nuestro, han establecido que la inflamación y la reactividad autoinmune desempeñan un papel en la retención de sodio y las respuestas hemodinámicas que provocan la hipertensión primaria. La hiperuricemia y las proteínas del estrés por calor (HSP), particularmente HSP70, están asociadas con la activación de la inmunidad innata que juega un papel en el desarrollo de la reactividad inflamatoria en pacientes hipertensos. Estudios clínicos han demostrado asociación entre la expresión de HSP70 y anticuerpos anti-HSP70 y la hipertensión arterial primaria Esta breve revisión tiene como objetivo examinar la interrelación entre la hiperuricemia y la sobreexpresión extracelular de HSP70 en la activación del inflamasoma, así como su probable papel central en la fisiopatología de la hipertensión primaria.

Expression profile in Leishmania major exposed to Staphylococcus aureus and group A beta-hemolytic Streptococcus

Background & objectives: The interaction of Leishmania spp. with microbiota inside the midgut vector has significant output in pathogenesis. This study aimed to identify the profile of Leishmania major gene expression of LACK, gp63, and hsp70 after exposure to Staphylococcus aureus and group A beta-hemolytic Streptococci (GABHS). Methods: Leishmania major (MRHO/IR/75/ER) promastigotes were exposed with S. aureus, with GABHS, and with both GABHS and S. aureus at 25°C for 72 h. The gene expression analysis of Lmgp63, Lmhsp70, and LmLACK was assessed using SYBR Green real-time PCR by ??Ct. All experiments were repeated in triplicate. Statistical analysis was done using two-way ANOVA. A P-value less than 0.05 was considered significant. Results: Lmgp63 was expressed in the group exposed to GABHS with 1.75-fold lower than the control group (p=0.000). The LmLACK had expression in both groups exposed with GABHS and GABHS with S. aureus with 2.8 and 1.33-fold more than the control group, respectively (p=0.000). The Lmhsp70 gene expression was reported in the group exposed with GABHS with relative quantification of 5.7-fold more than the control group. Interpretation & conclusion: This study showed that the important genes encoding LACK, gp63, and hsp70 changed their expression after exposure to the S. aureus and GABHS.

Identification of heat shock protein hsp70 family genes from Rana amurensis and its expression profiles upon infection / 生物工程学报

Heat shock proteins (HSPs) widely exist in all organisms, the structures of which are usually extraordinarily conservative. They are also well-known stress proteins that are involved in response to physical, chemical and biological stresses. HSP70 is an important member of the HSPs family. In order to study the roles of amphibians HSP70 during infection, the cDNA sequence of Rana amurensis hsp70 family genes were cloned by homologous cloning method. The sequence characteristics, three-dimensional structure and genetic relationship of Ra-hsp70s were analyzed by bioinformatics methods. The expression profiles under bacterial infection were also analyzed by real-time quantitative PCR (qRT-PCR). Expression and localization of HSP70 protein were tested by immunohistochemical techniques. The results showed that three conservative tag sequences of HSP70 family, HSPA5, HSPA8 and HSPA13, were found in HSP70. Phylogenetic tree analysis indicated four members are distributed in four different branches, and members with the same subcellular localization motif are distributed in the same branch. The relative expression levels of the mRNA of four members were all significantly upregulated (P < 0.01) upon infection, but the time for up-regulating the expression levels were diverse in different tissues. The immunohistochemical analysis showed that HSP70 was expressed to different degrees in the cytoplasm of liver, kidney, skin and stomach tissue. The four members of Ra-hsp70 family have ability to respond bacterial infection to varying degrees. Therefore, it was proposed that they are involved in biological processes against pathogen and play different biological functions. The study provides a theoretical basis for functional studies of HSP70 gene in amphibians.

Inflammation and damage-associated molecular patterns in major psychiatric disorders

Abstract Background Emerging evidence indicates that inflammation plays an important role as a mechanism underlying mental disorders. However, most of the research on inflammatory mechanisms focuses on serum levels of interleukins and very few studies have investigated molecules that initiate and expand innate immune pathways such as damage-associated molecular patterns (DAMPs). Objectives This study investigated the levels of DAMPs among patients diagnosed with major depressive disorder (MDD), bipolar disorder (BD) I and II, schizophrenia (SCZ), and generalized anxiety disorder (GAD). We quantified serum levels of heat shock proteins (HSPs) 70 and 60 and of S100 calcium-binding protein B (S100B). Methods Serum levels of HSP70, HSP60, and S100B were assessed in a sample of participants with psychiatric disorders (n = 191) and a control group (CT) (n = 59) using enzyme-linked immunosorbent assay (ELISA). Results Serum HSP70 concentrations were significantly higher in the MDD group compared to the CT, SCZ, and BD groups. The GAD group had higher concentrations of HSP70 than the SCZ group. Exploring associations with medications, lithium (p = 0.003) and clozapine (p = 0.028) were associated with lower HSP70 levels. Approximately 64% of the sample had DAMPs levels below the limits of detection indicated by the respective ELISA kit. Conclusion This was the first study to assess DAMPs levels in a transdiagnostic sample. Our preliminary findings suggest that HSP70 may be associated with MDD pathophysiology. Medications such as lithium and clozapine were associated with lower HSP70 levels in BD and SCZ groups, respectively. Therefore, it is worth mentioning that all participants were medicated and many psychotropic drugs exert an anti-inflammatory effect, possibly reducing the signs of inflammation.

Heat shock protein 70 inhibits hepatitis B virus replication by enhancing the deaminase activity of apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3B / 中华微生物学和免疫学杂志

Objective:To study the role and mechanism of heat shock protein 70 (HSP70) in apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3B (APOBEC3B)-mediated inhibition of hepatitis B virus(HBV) replication.Methods:The interaction between HSP70 and APOBEC3B was analyzed by co-immunopreciptation (Co-IP) and GST pull-down. After treating Huh7 cells with siHSP70 or HSP70 inhibitor VER155008 or overexpressing HSP70 in Huh7 cells, changes in the antiviral effect of APOBEC3B were detected by Southern blot and real-time PCR; the deaminase activity of APOBEC3B was tested by differential DNA denaturation PCR(3D-PCR) and clone sequencing.Results:HSP70 could bind to APOBEC3B. Overexpression of HSP70 promoted the deaminase activity and anti-HBV activity of APOBEC3B. On the contrary, knockdown of HSP70 or using HSP70 inhibitor VER155008 would attenuate the deaminase activity and anti-HBV activity of APOBEC3B.Conclusions:HSP70 could promote the anti-HBV activity of APOBEC3B by enhancing the deaminase activity of APOBEC3B.

Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM

ABSTRACT Background: Leishmaniasis is a vector-borne disease caused by a parasite protozoon from the genus Leishmania. Among the molecular techniques applied for detecting these parasites, real-time PCR with High Resolution Melting (PCR-HRM) proved advantageous since it simultaneously determines both the presence and species of the pathogen in one step, through amplification and later analysis of curves generated by melting temperature. Methods: Based on this molecular technique, the goal of this study was to estimate the PCR-HRM sensitivity for Leishmania spp. detection in different canine tissues by evaluating biological samples obtained from popliteal, submandibular, and pre-scapular lymph nodes, from bone marrow and ear pinnae of 28 stray dogs captured in the metropolitan area of Asunción (Paraguay). Results: The rk39 immunochromatographic test showed that 25/28 tested dogs (89%) presented antibodies against L. infantum. In 20/25 dogs that tested positive for rk39 (80%), it was possible to detect Leishmania spp. by PCR-HRM and determine that the species corresponded entirely to L. infantum. Regarding the analysis of different tissues, the parasite was detected in all popliteal lymph node samples, followed by high detection in submandibular (at 95%) and pre-scapular lymph nodes (at 90%), bone marrow (at 85%), and ear pinnae (at 85%). Conclusions: This study demonstrated that the use of real-time PCR-HRM using the molecular marker hsp70 was a highly sensitive method for simultaneously detecting and identifying Leishmania species in different tissues taken from infected dogs. In addition, the usefulness of ear pinnae as easily accessible tissue for molecular diagnosis was emphasized.

Estresse oxidativo e diminuição de HSP70 tecidual envolvidos na gênese da sepse: HSP70 como alvo terapêutico / Oxidative stress and decreased tissue HSP70 are involved in the genesis of sepsis: HSP70 as a therapeutic target

RESUMO A sepse é uma infecção sistêmica que acarreta disfunção múltipla dos órgãos. A HSP70 é uma proteína responsiva ao estresse celular, assim como o estresse oxidativo. Esta revisão da literatura buscou investigar a HSP70 e o estresse oxidativo quanto à fisiopatologia da sepse e ao papel da HSP70 como possível alvo terapêutico. A HSP70 exerce efeito protetor quando localizada na célula (iHSP70), e sua diminuição, assim como seu aumento no ambiente extracelular (eHSP70) e o estresse oxidativo, é um biomarcador de gravidade na sepse. Além disso, terapias que aumentam a iHSP70 ou o próprio tratamento com HSP70 promovem a melhora na sepse.

Abstract Sepsis is a systemic infection that causes multiple organ dysfunction. HSP70 is a protein responsive to cell stress, in particular oxidative stress. Therefore, this literature review sought to investigate the roles of HSP70 and oxidative stress in the pathophysiology of sepsis and the possibility of HSP70 as a therapeutic target. HSP70 exerts a protective effect when located in cells (iHSP70), and its decrease, as well as its increase in the extracellular environment (eHSP70), under oxidative stress is a biomarker of sepsis severity. In addition, therapies that increase iHSP70 and treatment with HSP70 promote sepsis improvement.

Aqueous humor heat-shock protein 70, periostin, and irisin levels in patients with pseudoexfoliation syndrome / Níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso em pacientes com pseudoexfoliação

ABSTRACT Purpose: To measure humor heat-shock protein 70, periostin, and irisin levels in patients with pseudoexfoliation syndrome and cataract (without glaucoma), and compare them with those of patients with cataract but without pseudoexfoliation. Methods: We examined 31 eyes of 31 patients with pseudoexfoliation and cataract (without glaucoma) and 30 eyes of 30 patients with cataract. We collected aqueous humor samples from all patients at the time of cataract surgery through a limbal paracentesis via a 25-gauge cannula mounted on a tuberculin syringe that received 100 to 150 µL of aqueous humor. We measured levels of aqueous humor Heat shock protein 70, periostin, and irisin using enzyme-linked immunosorbent assay methods. Results: The age (p=0.221) and gender (p=0.530) means were similar between the pseudoexfoliation and control groups. The mean Heat shock protein 70 level (29.22 ± 9.46 ng/mL; 17.88-74.46) in the pseudoexfoliation group was significantly higher than that in the control group (19.03 ± 7.05 ng/mL; 9.93-35.52; p<0.0001). The mean periostin level was significantly higher (6017.32 ± 1271.79 pg/mL; 3787.50-10803.57) in the pseu doexfoliation group than that in the control group (4073.63 ± 1422.79 pg/mL; 2110.44-7490.64; p<0.0001). The mean irisin level (53.77 ± 10.19 ng/mL; 29.46-71.16) was significantly higher than that in the control group (39.29 ± 13.58 ng/mL; 19.41-70.56; p<0.0001). Conclusions: Heat shock protein 70, periostin, and irisin levels increase in the aqueous humor of patients with pseudoexfoliation without glaucoma.

RESUMO Objetivo: Comparar os níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso de pacientes com pseudoexfoliação com catarata sem glaucoma e compará-los com pacientes com catarata sem pseudoexfoliação. Métodos: Trinta e um olhos de 31 pacientes com pseudoexfoliação com catarata sem glaucoma e 30 olhos de 30 indivíduos com catarata foram incluídos neste estudo. Amostras de humor aquoso foram coletadas de todos os pacientes no momento da cirurgia de catarata e obtidas através de uma paracentese límbica por meio de uma cânula de calibre 25 acoplada a uma seringa com tuberculina. Foram coletados 100 a 150 µL de humor aquoso. Os níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso foram medidos usando o método de ensaio imunossorvente ligado a enzima. Resultados: A média da idade (p=0,221) e sexo (p=0,530) foram semelhantes entre os grupos pseudoexfoliação e controle. Os níveis médios de proteína de choque térmico 70 foram 29,22 ± 9,46 ng/mL (17,88-74,46) e 19,03 ± 7,05 ng/ mL (9,93-35,52) nos grupos pseudoexfoliação e controle, respectivamente. Os níveis de proteína de choque térmico 70 foram maiores no grupo pseudoexfoliação (p<0,0001). O nível médio de periostina foi de 6017,32 ± 1271,79 pg/mL (3787,50-10803,57) no grupo pseudoexfoliação e 4073,63 ± 1422,79 pg/mL (2110,44-7490,64) no grupo controle. O nível médio de periostina também foi maior no grupo pseudoexfoliação (p<0,0001). Os níveis médios de irisina foram 53,77 ± 10,19 ng/mL (29,46-71,16) e 39,29 ± 13,58 ng/mL (19,41-70,56) nos grupos pseudoexfoliação e controle, respectivamente. O nível médio de irisina foi maior no grupo pseudoexfoliação do que no grupo controle (p<0,0001). Conclusões: Os níveis de proteína de choque térmico 70, de periostina e de irisina aumentam no humor aquoso de pacientes com pseudoexfoliação sem glaucoma.

Quinacrine causes apoptosis in human cancer cell lines through caspase-mediated pathway and regulation of small-GTPase

Quinacrine (QC), an FDA-approved anti-malarial drug, has shown to have anticancer activities. Due to its‘shotgun’ nature, QC has become an inevitable candidate for combination chemotherapy. There is lack of studyof the molecular interplay between colorectal cancer (CRC) microenvironment and its metastasis. In this study,we focused on the differential anti-cancerous effect of QC on two different human cancer cell lines, HCT 116and INT 407. Results suggest that cytotoxicity increased in both the cell lines with an increase in QCconcentration. The expression patterns of small-GTPases and caspases were altered significantly in QC-treatedcells compared to non-treated cells. HSP70 and p53 showed comparable differences in the expression pattern.The wound-healing assay showed an increase in the denuded zone, with an increase in the concentration ofQC. The formation of apoptotic nuclei increased with a rise in the concentration of QC in both the cell lines.The decrease and increase in caspase 9 and caspase 3 expression respectively were studied, confirmingapoptosis by the extrinsic pathway

Evidence of a sylvatic enzootic cycle of Leishmania infantum in the State of Amapá, Brazil

Abstract INTRODUCTION: Leishmania infantum was considered to be absent from Amapá until 2017 when canine infection was detected. However, there is a lack of knowledge about which reservoir species are involved in transmission in this region. METHODS: Between 2014 and 2016, 86 samples from wild mammals and 74 from domestic dogs were collected in Wajãpi Indigenous Territory and were tested for the presence of deoxyribonucleic acid (DNA) of Leishmania. RESULTS: The DNA of Le. infantum was detected in two rodent samples, Dasyprocta sp. and Proechimys cuvieri. CONCLUSIONS: This is the first evidence characterizing a sylvatic transmission cycle of Le. infantum in the State of Amapá.

Effects of Liuwei Dihuang Decoction (Yukmijihwang-tang) on Physical Fatigue by Regulating Neurotransmitters in Brain / 中国结合医学杂志

OBJECTIVE@#To study the effect of Liuwei Dihuang Decoction () or Yukmijihwangtang (YJT) on endurance exercise by in vivo experiment.@*METHODS@#ICR mice were randomly divided into the control group (distilled water) and the YJT groups (1, 10, 100 mg/kg), 5 animals per group. YJT and distilled water were orally administered. The anti-fatigue effect of YJT was evaluated by open fifiled test (OFT), forced swimming test (FST), and tail suspension test (TST).@*RESULTS@#In the OFT, YJT signifificantly increased the total movement distance in a dose-dependent manner. Additionally, treatment with YJT signifificantly decreased immobility time in the FST and the TST. Various neurotransmitters such as norepinephrine (NE), serotonin (5-HT), dopamine (DA) levels were increased by FST. Administration of YJT down-regulated the expression levels of NE, 5-HT, 5-hydroxyindole-acetic acid (5-HIAA), and DA in the brain stem and hypothalamus of mice. Moreover, protein expression of HSP70 in mice liver and heart muscles was signifificantly increased in the YJT groups.@*CONCLUSIONS@#YJT could ameliorate fatigue and enhance exercise tolerance through suppressing of brain monoamines including NE, 5-HT, 5-HIAA, and DA in FST mice model.

The expression of and preoperative correlation between heat-shock protein 70, euroscore, and lactate in patients undergoing cabg with cardiopulmonary bypass

Abstract Objetive: Coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) improved symptoms and increased survival and quality of life in patients with coronary artery disease. However, it should be the main cause of a complex organic systemic inflammatory response that greatly contributes to several postoperative adverse effects. Methods: We aimed to evaluate heat-shock protein 70 (HSP 70) expression as a morbimortality predictor in patients with preserved ventricular function undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) and to determine their association with the lactate as a marker of tissue hypoperfusion and the EuroSCORE risk score. This is a prospective, observational study including 46 patients and occurring between May and July 2016. Patients without ventricular dysfunction undergoing myocardial revascularization with extracorporeal circulation were included. They were divided into (1) complicated and (2) uncomplicated postoperative evolution groups. EuroSCORE, lactate levels, and HSP 70 expression and their correlations were determined. Results: Statistical analysis showed that the group with complicated evolution had higher EuroSCORE values than the other group. HSP 70 protein levels were significantly increased in the group with uncomplicated evolution and showed similar results. According to our results, HSP family proteins may be independent predictors of uncomplicated evolution in patients without ventricular dysfunction undergoing CABG with CPB. Conclusion: HSP 70 should be a good discriminator and protection marker for complications in cardiac surgery.

Effect of moxibustion on expressions of HSP70 mRNA and protein in gastric cancer-bearing rats / 针灸推拿医学（英文版）

Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70 (HSP70) in gastric cancer-bearing rats. Methods: A total of 40 healthy Sprague-Dawley (SD) rats were adaptively fed for one week. The gastric cancer model was prepared by Walker-256 cancer tissue transplantation. After 7 d, 10 rats were randomly selected to verify the successful modeling, and the remaining 30 rats were divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. After enrollment, the moxibustion group received suspended moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36), (the first group of acupoints) on the 1st day, and suspended moxibustion at bilateral Pishu (BL 20) and Weishu (BL 21), (the second group of acupoints) on the 2nd day, 20 min each time, once a day. Moxibustion was alternately performed every other day at the two groups of acupoints for 21 d. From the day of enrollment, rats in the infrared group were irradiated with the infrared radiation at the stomach area on the 1st day, and at the T12-T13 interspinous region on the 2nd day, 20 min each time, once a day, and the two locations were alternately irradiated every other day for 21 d. During the treatment, rats in the model group were intervened by grasping and fixation without treatment. At the end of the treatment, blood was collected from the inner eye orbit, and the HSP70 expression in peripheral blood was determined by enzyme linked immunosorbent assay (ELISA). Rats were sacrificed, the tumor volume and growth inhibition rate were measured. The position and changes of HSP70 in gastric cancer were observed by streptavidin-perosidase (SP); HSP70 protein expression was determined by ELISA; HSP70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. Results: In comparison of the model group, the volume growth of the gastric cancer in the moxibustion group was significantly restricted (P＜0.01); the volume growth inhibition rate in the moxibustion group was 37.93%; the HSP70 expression in peripheral blood and the cancer tissues was significantly increased (both P＜0.01); the expression of HSP70 mRNA and HSP70 content in gastric tumor were both obviously increased in the moxibustion group (P＜0.01); and a large amount of HSP70 was released to the outside of cancer cells in the moxibustion group. In comparison of the model group, the volume growth of the gastric cancer in the infrared group was slightly restricted (P＜0.05) with a volume growth inhibition rate of 15.89%; the HSP70 expression in the infrared group was increased significantly in peripheral blood (P＜0.01) and in the gastric cancer tissues (P＜0.05); more HSP70 was released outside of the cancer cells in the infrared group. In comparison of the infrared group, the volume growth of gastric cancer was more restricted in the moxibustion group (P＜0.05), and the HSP70 expression in the gastric cancer tissues was also higher (P＜0.05); more HSP70 was released outside of the cancer cells in the moxibustion group. Conclusion: Moxibustion and infrared treatment inhibit the gastric cancer growth in the gastric cancer-bearing rats, up-regulate the HSP70 expression in gastric cancer tissues, and promote the production and extracellular release of HSP70, and the effect of moxibustion is more obvious.

Heat shock protein 70 in nuclear translocation involved in DNA repair during ischemia-reperfusion injury / 中华普通外科杂志

Objective To explore the mechanism of HSP70 on cell cycle regulation in hepatic IR (ischemia-reperfusion) injury.Methods SD rats were randomly divided into HSP70 inhibitor group (H-/P+),heat shock group (H+/P+),PARP-1 inhibitory group (H +/P-),IR group (PC) and negative control group (NC),respectively.After the IR model was induced,the liver specimen underwent IHC staining to observe the changes of the PARP-1 expression;Co-immunoprecipitation was used to detect the binding of HSPT0 with PARP-1.Results H +/P-was significantly different from H +/P +,H-/P +,PC,NC (P ＜ 0.01);Immunoprecipitation suggested that HSP70 entered into the nucleus to bind PARP-1,and immunofluorescence imaging analysis demonstrated both HSP70 and CyclinD1 expressed at the same timeline.Conclusion Under reversible hepatic ischemia-reperfusion injury,HSP70 enters the nucleus and binds to PARP-1,negatively regulates G2/M phase,blocks cells for DNA replication and recombination,blocks its entry into mitosis,repairs damaged DNA chain;Liver ischemia-reperfusion positively regulates the G1/S phase,promoting hepatocyte regeneration and liver function compensation.

Effects of heme oxygenase-1 upregulation on isoproterenol-induced myocardial infarction

The present study was designed to examine the effect of heme oxygenase-1 (HO-1) induction by cobalt protoporphyrin (CoPP) on the cardiac functions and morphology, electrocardiogram (ECG) changes, myocardial antioxidants (superoxide dismutase [SOD] and glutathione [GSH]), and expression of heat shock protein (Hsp) 70 and connexin 43 (Cx-43) in myocardial muscles in isoproterenol (ISO) induced myocardial infarction (MI). Thirty two adult male Sprague Dawely rats were divided into 4 groups (each 8 rats): normal control (NC) group, ISO group: received ISO at dose of 150 mg/kg body weight intraperitoneally (i.p.) for 2 successive days; ISO + Trizma group: received (ISO) and Trizma (solvent of CoPP) at dose of 5 mg/kg i.p. injection 2 days before injection of ISO, with ISO at day 0 and at day 2 after ISO injections; and ISO + CoPP group: received ISO and CoPP at a dose of 5 mg/kg dissolved in Trizma i.p. injection as Trizma. We found that, administration of ISO caused significant increase in heart rate, corrected QT interval, ST segment, cardiac enzymes (lactate dehydrogenase, creatine kinase-muscle/brain), cardiac HO-1, Hsp70 with significant attenuation in myocardial GSH, SOD, and Cx-43. On the other hand, administration of CoPP caused significant improvement in ECG parameters, cardiac enzymes, cardiac morphology; antioxidants induced by ISO with significant increase in HO-1, Cx-43, and Hsp70 expression in myocardium. In conclusions, we concluded that induction of HO-1 by CoPP ameliorates ISO-induced myocardial injury, which might be due to up-regulation of Hsp70 and gap junction protein (Cx-43).

UPP pathway involves in regulating degradation of hyperphosphorylated tau protein in aluminum-induced N2a cells / 中国职业医学

OBJECTIVE: To explore the mechanism of ubiquitin-proteasome pathway(UPP) in the degradation of hyperphosphorylated tau protein in aluminum-induced mouse neuroblastoma N2 a cells. METHODS: N2 a cells in logarithmic growth period were randomly divided into control group and MG132 group. Cells in control group were exposed to concentrations of 0 or 1 mmol/L aluminum chloride for 24 hours. Cells in MG132 group were pretreated with MG132 at a concentration of 5 μmol/L for 6 hours, then exposed to concentrations of 0 or 1 mmol/L aluminum chloride for 24 hours. After exposure, the cells were collected. Western blotting was used to detect the relative expression of tau-5, P-tau181, P-tau231, P-tau262, P-tau396, heat shock protein 70(Hsp70) and carboxyl terminus of the Hsp70-interacting protein(CHIP). The ubiquitin relative expression was detected by enzyme-linked immunosorbent assay. RESULTS: The results of factorial analysis showed that the relative expression of tau-5, P-tau231, P-tau262, P-tau396, CHIP, Hsp70 and ubiquitin in N2 a cells were statistically significant in the main effect and interaction effect of aluminum chloride and MG132 treatment(P<0.05). Both in the control group and MG132 group, the relative expression of tau-5, P-tau231, P-tau262, P-tau396, CHIP, Hsp70 and ubiquitin in N2 a cells exposed to 1 mmol/L aluminum chloride increased(P<0.05) when compared with the N2 a cells without exposed to aluminum chloride. No matter aluminum chloride exposed or not, the relative expression of tau-5, P-tau231, P-tau262, P-tau396, CHIP, Hsp70 and ubiquitin in N2 a cells of MG132 group was higher than that of control group(P<0.05). CONCLUSION: UPP is involved in the regulation of hyperphosphorylated tau protein by proteasome degradation in aluminum-induced N2 a cells. UPP mainly regulates P-tau231, P-tau262, and P-tau396 sites. CHIP and Hsp70 played an important role in the UPP pathway.

Effect of exercise training on heat shock protein 70 expression during endotoxin-induced acute lung / 中华麻醉学杂志

Objective To evaluate the effect of exercise training on heat shock protein 70 (HSP70) expression during endotoxin-induced acute lung injury (ALI) in rats.Methods Thirty-two SPF healthy male Sprague-Dawley rats,aged 8 weeks,weighing 175-220 g,were divided into 4 groups (n=8 each) using a random number table method:control group (group C),group ALI,low-intensity exercise training group (group ET1) and high-intensity exercise training group (group ET2).The rats in ET1 and ET2 groups received 2-and 4-week treadmill exercise training before establishing the ALI model,while the rats in C and ALI groups received no training.ALI was induced by intravenously injecting 5 mg/kg lipopolysaccharide via the tail vein in ALI,ET1 and ET2 groups,and the equal volume of normal saline was given instead in group C.The animals were sacrificed,and the lungs were harvested for microscopic examination of the pathological changes of lung tissues which were also scored and for determination of wet to dry weight ratio (W/D ratio),concentrations of total protein,interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) in bronchoalveolar lavage fluid (BALF),and expression of HSP70 and nuclear factor kappa B (NF-κB) in lung tissues by Western blot.Results Compared with group C,the W/D ratio and pathological changes of lung tissues were significantly increased,the concentrations of total protein,IL-1 β and TNF-α in BALF were increased,the expression of NF-κB was up-regulated (P＜0.05),and no significant change was found in HSP70 expression in group ALI(P＞0.05).Compared with group ALI,the W/D ratio and pathological changes of lung tissues were significantly decreased,the concentrations of total protein,IL-1β and TNF-α in BALF were decreased,the expression of HSP70 was up-regulated,and the expression of NF-κB was down-regulated in ET1 and ET2 groups (P＜0.05).Conclusion Exercise training can attenuate the endotoxin-induced ALI through relieving the inflammatory responses,which may be related to up-regulating HSP70 expression in the lung of rats.

Effects of chromium picolinate supplementation on growth performance, small intestine morphology and antioxidant status in ducks under heat stress conditions / Efectos de la suplementación con picolinato de cromo en el rendimiento de crecimiento, morfología del intestino delgado y estado antioxidante en patos bajo condiciones de estrés calórico

SUMMARY: The experiment was conducted to evaluate the effects of dietary supplemental chromium (Cr) on growth performance, meat quality, intestinal morphology, mucosa Hsp70 mRNA expression and antioxidant status of ducks reared under heat stress conditions. All ducks were randomly divided into three treatment groups, respectively, control group (Control, 23 ± 2 °C), heat stress group (HS, 32 ±2 °C), Cr picolinate group (CrPic, 32 ± 2 °C, 0.2 mg Cr/kg). Feed and distilled-deionized water were available ad libitum for an experimental phase of 35 days. Samples were collected on the day 14, 21 and 35 to determine biological and hematological values. Results showed that heat stress or dietary supplemental Cr both didn't have distinct influence on growth performance (P>0.05), compared to controls. Ducks fed 0.2 mg Cr/kg diet had greater ultimate pH (pHu)(P<0.05) than HS group. At day 14, the ratio of villus height to crypt depth (V/C) in CrPic group significantly increased (P<0.05) than that of HS group in jejunum. Heat stress remarkably increased Hsp70 mRNA expression in jejunum compared with controls (P<0.05). While the expression of Hsp70 mRNA in CrPic group was significantly decreased compared with HS (P<0.05). At day 21, the V/C of ileum in CrPic group significantly increased compared with HS group (P<0.05). Serum SOD levels in CrPic group were significantly higher than those in HS group (P<0.05). At day 35, Hsp70 mRNA expression and serum T-SOD levels in CrPic group significantly increased compared with controls (P<0.05). T-AOC in HS group significantly decreased compared with controls (P<0.05). Results indicate that dietary Cr supplementation doesn't influence ducks' growth performance, but has a positive effect on meat quality, small intestine morphology, also regulates Hsp70 mRNA expression under heat stress conditions, and enhances the antioxidant status.

RESUMEN: Se evaluó los efectos del cromo (Cr) dietético suplementario sobre el rendimiento del crecimiento, la calidad de la carne, la morfología intestinal, la expresión del ARNm Hsp70 en la mucosa y el estado antioxidante de los patos criados bajo condiciones de estrés por calor. Todos los patos se dividieron aleatoriamente en tres grupos: grupo control (control, 23 ± 2 °C), grupo de estrés térmico (HS, 32 ± 2 °C) y grupo de picolinato de Cr (CrPic, 32 ± 2 °C, 0,2 mg Cr / kg). El alimento y el agua desionizada destilada estuvieron disponibles ad libitum durante la fase experimental de 35 días. Las muestras se recogieron los días 14, 21 y 35 para determinar los valores biológicos y hematológicos. Los resultados mostraron que el estrés térmico o la suplementación dietética de Cr no tuvieron una influencia distinta en el rendimiento del crecimiento (P> 0,05), en comparación con los controles. Los patos alimentados con 0,2 mg de Cr / kg de dieta tuvieron un mayor pH final (pHu) (P <0,05) que el grupo HS. En el día 14, la relación de la altura de las vellosidades a la profundidad de la cripta (V / C) en el grupo CrPic aumentó significativamente (P <0,05) en relación a la del grupo de HS en el yeyuno. El estrés por calor incrementó notablemente la expresión del ARNm de Hsp70 en el yeyuno en comparación con los controles (P <0,05). Mientras que la expresión del ARNm de Hsp70 en el grupo CrPic se redujo significativamente en comparación con HS (P <0,05). En el día 21, la relación V / C del íleon en el grupo CrPic aumentó significativamente en comparación con el grupo HS (p <0,05). Los niveles séricos de SOD en el grupo CrPic fueron significativamente más altos que los del grupo HS (P <0,05). En el día 35, la expresión de ARNm de Hsp70 y los niveles séricos de T-SOD en el grupo CrPic aumentaron significativamente en comparación con los controles (P <0,05). T-AOC en el grupo HS disminuyó significativamente en comparación con los controles (P <0,05). Los resultados indican que la suplementación dietética de Cr no influye en el rendimiento de crecimiento de los patos, pero tiene un efecto positivo en la calidad de la carne, en la morfología del intestino delgado, y también regula la expresión de ARNm de Hsp70 en condiciones de estrés calórico y mejora el estado antioxidante.

HSP70 from the Antarctic sea urchin Sterechinus neumayeri: molecular characterization and expression in response to heat stress

Abstract Background: Heat stress proteins are implicated in stabilizing and refolding denatured proteins in vertebrates and invertebrates. Members of the Hsp70 gene family comprise the cognate heat shock protein (Hsc70) and inducible heat shock protein (Hsp70). However, the cDNA sequence and the expression of Hsp70 in the Antarctic sea urchin are unknown. Methods: We amplified and cloned a transcript sequence of 1991 bp from the Antarctic sea urchin Sterechinus neumayeri, experimentally exposed to heat stress (5 and 10 °C for 1, 24 and 48 h). RACE-PCR and qPCR were employed to determine Hsp70 gene expression, while western blot and ELISA methods were used to determine protein expression. Results: The sequence obtained from S. neumayeri showed high identity with Hsp70 members. Several Hsp70 family features were identified in the deduced amino acid sequence and they indicate that the isolated Hsp70 is related to the cognate heat shock protein type. The corresponding 70 kDa protein, called Sn-Hsp70, was immune detected in the coelomocytes and the digestive tract of S. neumayeri using a monospecific polyclonal antibody. We showed that S. neumayeri do not respond to acute heat stress by up-regulation of Sn-Hsp70 at transcript and protein level. Furthermore, the Sn-Hsp70 protein expression was not induced in the digestive tract. Conclusions: Our results provide the first molecular evidence that Sn-Hsp70 is expressed constitutively and is noninduced by heat stress in S. neumayeri.

Mechanism of heart injury in brain death rabbits / 中华器官移植杂志

Objective To study the mechanism of brain death-induced heart damage by observing the change patterns of morphological damage to the heart and related inflammatory factors after brain death and provide the experimental basis for heart transplantation by brain-dead donor.Methods The 30 rabbits were equally divided into two groups by the random digital table method:sham-operation group and brain death group.The rabbit brain death model was established in the brain death group,and the sham-operation group was given slow intracranial pressure.The rest treatments in the two groups were the same.At 2nd,6th and 8th h after operation,blood pressure,heart rate and respiratory rate were recorded.The damage of heart tissues was observed by HE staining.The plasma concentrations of IL-1,IL-6 and IL-8 were tested by ELISA.The expression of some inflammatory factors in heart issues was detected by RT-PCR and immunohistochemistry.Results At 8 h after brain death,there was no signifiant difference in blood pressure and heart rate between two groups (P＞0.05).The damage of heart issues in the brain death group was more serious than in the shamoperation group.With the prolongation of brain death,the plasma concentrations of IL-6 and IL-8 increased significantly in the brain death group (P＜0.05),but the concentration of IL-1β showed no siginificant difference between the two groups at 2 h after brain death (P＜0.05).Besides,the expression of HSP27 and HSP70 mRNA as well as the protein expression of ICAM and NF-κB was significantly increased in the brain death group as compared with that in the sham-operation group (P＜0.05).Conclusion With the prolongation of brain death time,the inflammatory factors in the heart tissues and plasma interleukin were increased,suggesting the inflammatory reaction occurs in donor heart under the condition of brain death,which influences the quality of donor in the heart transplantation.