Role of heat shock protein 70 in regulation of anti-inflammatory response to curcumin in 3T3-L1 adipocytes.

BACKGROUND/OBJECTIVES: Curcumin is a well-known phytochemical with anti-inflammatory effects. Heat shock protein (HSP) 70, an intracellular chaperone, inhibits proinflammatory signaling activation. Although curcumin has been shown to induce HSP70 expression in various cell types, whether HSP70 mediates the anti-inflammatory effects of curcumin in mature adipocytes remains unclear. MATERIALS/METHODS: To assess the role of HSP70 in regulating the anti-inflammatory response to curcumin in adipocytes, fully differentiated 3T3-L1 adipocytes were treated with curcumin, lipopolysaccharide (LPS), and/or the HSP70 inhibitor pifithrin-µ (PFT-µ). The expression levels of HSP70 and proinflammatory cytokines were then measured. RESULTS: Curcumin upregulated HSP70 expression at both protein and mRNA levels and attenuated LPS-induced Il6, Ptx3, and Ccl2 mRNA upregulation. PFT-µ tended to exacerbate the LPS-induced upregulation of Il6, Ptx3, Ccl2, and Tnfa mRNA expression. However, on curcumin pretreatment, the tendency of PFT-µ to upregulate LPS-induced proinflammatory cytokine expression decreased or disappeared. CONCLUSION: These results indicate that HSP70 is involved in the regulation of inflammatory responses but may not be crucial for the anti-inflammatory effects of curcumin in 3T3-L1 adipocytes.

Jmjd4 Facilitates Pkm2 Degradation in Cardiomyocytes and Is Protective Against Dilated Cardiomyopathy.

BACKGROUND: A large portion of idiopathic and familial dilated cardiomyopathy (DCM) cases have no obvious causal genetic variant. Although altered response to metabolic stress has been implicated, the molecular mechanisms underlying the pathogenesis of DCM remain elusive. The JMJD family proteins, initially identified as histone deacetylases, have been shown to be involved in many cardiovascular diseases. Despite their increasingly diverse functions, whether JMJD family members play a role in DCM remains unclear. METHODS: We examined Jmjd4 expression in patients with DCM, and conditionally deleted and overexpressed Jmjd4 in cardiomyocytes in vivo to investigate its role in DCM. RNA sequencing, metabolites profiling, and mass spectrometry were used to dissect the molecular mechanism of Jmjd4-regulating cardiac metabolism and hypertrophy. RESULTS: We found that expression of Jmjd4 is significantly decreased in hearts of patients with DCM. Induced cardiomyocyte-specific deletion of Jmjd4 led to spontaneous DCM with severely impaired mitochondrial respiration. Pkm2, the less active pyruvate kinase compared with Pkm1, which is normally absent in healthy adult cardiomyocytes but elevated in cardiomyopathy, was found to be drastically accumulated in hearts with Jmjd4 deleted. Jmjd4 was found mechanistically to interact with Hsp70 to mediate degradation of Pkm2 through chaperone-mediated autophagy, which is dependent on hydroxylation of K66 of Pkm2 by Jmjd4. By enhancing the enzymatic activity of the abundant but less active Pkm2, TEPP-46, a Pkm2 agonist, showed a significant therapeutic effect on DCM induced by Jmjd4 deficiency, and heart failure induced by pressure overload, as well. CONCLUSIONS: Our results identified a novel role of Jmjd4 in maintaining metabolic homeostasis in adult cardiomyocytes by degrading Pkm2 and suggest that Jmjd4 and Pkm2 may be therapeutically targeted to treat DCM, and other cardiac diseases with metabolic dysfunction, as well.

Comparison of allosteric signaling in DnaK and BiP using mutual information between simulated residue conformations.

The heat shock protein 70 kDa (Hsp70) chaperone system serves as a critical component of protein quality control across a wide range of prokaryotic and eukaryotic organisms. Divergent evolution and specialization to particular organelles have produced numerous Hsp70 variants which share similarities in structure and general function, but differ substantially in regulatory aspects, including conformational dynamics and activity modulation by cochaperones. The human Hsp70 variant BiP (also known as GRP78 or HSPA5) is of therapeutic interest in the context of cancer, neurodegenerative diseases, and viral infection, including for treatment of the pandemic virus SARS-CoV-2. Due to the complex conformational rearrangements and high sequential variance within the Hsp70 protein family, it is in many cases poorly understood which amino acid mutations are responsible for biochemical differences between protein variants. In this study, we predicted residues associated with conformational regulation of human BiP and Escherichia coli DnaK. Based on protein structure networks obtained from molecular dynamics simulations, we analyzed the shared information between interaction timelines to highlight residue positions with strong conformational coupling to their environment. Our predictions, which focus on the binding processes of the chaperone's substrate and cochaperones, indicate residues filling potential signaling roles specific to either DnaK or BiP. By combining predictions of individual residues into conformationally coupled chains connecting ligand binding sites, we predict a BiP specific secondary signaling pathway associated with substrate binding. Our study sheds light on mechanistic differences in signaling and regulation between Hsp70 variants, which provide insights relevant to therapeutic applications of these proteins.

The Change in Circadian Rhythms in P301S Transgenic Mice is Linked to Variability in Hsp70-related Tau Disaggregation.

Circadian disruption often involves a neurodegenerative disorder, such as Alzheimer's disease or frontotemporal dementia, which are characterized by intraneuronal tau accumulations. The altered sleep pattern and diurnal rhythms in these disorders are the results of tau pathology. The circadian disturbance in reverse is thought to develop and potentially aggravate the condition. However, the underlying mechanism is not fully understood. In this study, perturbed oscillations in BMAL1 , the core clock gene, were observed in P301S tau transgenic mice. Tau fractionation analysis of the hippocampus revealed profound fluctuations in soluble and insoluble tau protein levels that were in opposite directions to each other according to zeitgeber time. Interestingly, a diurnal oscillation was detected in the heat shock 70 kDa protein 1A (Hsp70) chaperone that was in-phase with soluble tau but out-of-phase with insoluble tau. Tau protein levels decreased in the soluble and insoluble fractions when Hsp70 was overexpressed in HEK293T cells. Transfection of the BMAL1 carrying vector was continual with the increase in Hsp70 expression and diminished tau protein levels, and it was effectively attenuated by the knockdown of Hsp70, suggesting that Bmal1 could modulate tau protein by Hsp70. Our results suggest that altered circadian oscillations affect tau status and solubility by modulating Hsp70 expression in an experimental model of tau pathology. These findings suggest Hsp70 as a possible pathogenic link between circadian disruption and aggravations of tau pathology.

The Chaperone Protein GRP78 Promotes Survival and Migration of Head and Neck Cancer After Direct Radiation Exposure and Extracellular Vesicle-Transfer.

Background and Purpose: Increased levels of the chaperone protein GRP78 have been implicated in poorer outcomes of cancer therapy. We have therefore explored the functional connection between the expression of GRP78 and the development of radioresistance and metastatic behavior in HNSCC. Material and Methods: The association between gene expression of GRP78 and survival in HNSCC patients was examined using the TCGA database. The influence of ionizing radiation on the GRP78 levels in HNSCC cell lines, their secreted extracellular vesicles (EV) and non-irradiated EV-recipient cells was investigated by Western Blot and FACS. The consequences of chemical inhibition or experimental overexpression of GRP78 on radioresistance and migration of HNSCC cells were analyzed by clonogenic survival and gap closure assays. Results: Elevated levels of GRP78 RNA in HNSCC correlated with poorer overall survival. Radiation increased GRP78 protein expression on the surface of HNSCC cell lines. Experimental overexpression of GRP78 increased both radioresistance and migratory potential. Chemical inhibition of GRP78 impaired cell migration. EVs were identified as a potential source of increased GRP78 content as elevated levels of surface GRP78 were found in EVs released by irradiated cells. These vesicles transferred GRP78 to non-irradiated recipient cells during co-cultivation. Conclusions: We have identified the chaperone protein GRP78 as a potential driver of increased radioresistance and motility in HNSCC. The uptake of GRP78-rich EVs originating from irradiated cells may contribute to a poorer prognosis through bystander effects mediated by the transfer of GRP78 to non-irradiated cells. Therefore, we consider the chaperone protein GRP78 to be an attractive target for improving radiotherapy strategies.

Evaluation of Salivary Level of Heat Shock Protein 70 in Patients with Chronic Periodontitis.

STATEMENT OF THE PROBLEM: Traditional clinical criteria are usually not sufficient for determining the sites of active periodontal disease, monitoring the response to treatment, or measuring the susceptibility to future disease development. Past studies have shown that heat shock protein 70 (HSP70) are involved in the etiology of periodontal disease. PURPOSE: The aim of this study was to evaluate the level of HSP70 in saliva of patients with chronic periodontitis (CP). MATERIALS AND METHOD: In our case-control study, the saliva samples of 45 patients with CP and 45 age- and sex-matched healthy subjects were collected. Salivary HSP70 was measured by enzyme-linked immunosorbent assay method. The results were analyzed by statistical tests using SPSS 16 and the statistically significant difference was set at p< 0.05. RESULTS: In this study, the mean salivary HSP70 level was 2.81±0.61ng/ml in the patient group and 1.96±0.77ng/ml in the healthy group, with a significant difference (p< 0.05). In addition, the results of spearman correlation analysis showed a positive correlation between salivary HSP 70 and clinical periodontal index. CONCLUSION: The results of this study showed that the salivary HSP70 level in patients with CP is higher than that in healthy subjects. As a result, salivary HSP70 might be considered as a marker in the pathogenesis of CP.

Photobiomodulation therapy on expression of HSP70 protein and tissue repair in experimental acute Achilles tendinitis.

The aim of the present study was to investigate the effects of photobiomodulation (PBM) therapy on the expression of heat shock protein 70 (HSP70) and tissue repair in an experimental model of collagenase-induced Achilles tendinitis. Thirty Wistar rats (aged 12 weeks) were randomly distributed among control group (n = 8), tendinitis group (n = 11), and LED group (n = 11). Tendinitis was induced in the tendinitis and LED groups through a peritendinous injection of collagenase (100 µl). The LED group animals received the first irradiation 1 h after injury. A 630 ± 20 nm, 300-mW continuous wave light-emitting diode (LED), spot size 1 cm2, was placed in contact with the skin. One point over the tendon was irradiated for 30 s, delivering 9 J (9 J/cm2). LED irradiation was performed once daily for 7 days, with the total energy delivered being 63 J. The tendons were surgically removed and expression of the HSP70 protein was calculated using semi-quantitative analyses of immunohistochemistry (HSCORE). Number of fibroblasts and amount of collagen were measured using histological and histochemical analyses. An increase in the mean HSCORE for HSP70, in the number of fibroblasts, and in the amount of collagen were found in the LED group compared with those in the tendinitis and control group (P ≤ 0.05). PBM therapy increased the expression of the HSP70, number of fibroblasts, and amount of collagen in the acute Achilles tendinitis in rats.

Indolyl- and Pyrrolylazine Derivatives Cause the Accumulation of Heat Shock Protein Hsp70 in Sh-Sy5Y Human Neuroblastoma Cells.

The heat shock protein Hsp70 is involved in cell defense from various types of stress, including the proteotoxic stress, which occurs during the development of many neurodegenerative diseases. This work presents data on the detection of small molecules, derivatives of indolyl- and pyrrolylazines, which can activate the synthesis of Hsp70 and cause its accumulation in the cell. The toxicity level of the new Hsp70 synthesis inducers was evaluated, and the safety of these compounds was demonstrated in experiments on SH-SY5Y neuroblastoma cell line. Derivatives of indolyl- and pyrrolylazines presented in this work can be potential therapeutic agents in models of neurodegenerative diseases that should be studied in more detail.

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.

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).

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 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.

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).

An evaluation of the effect of saffron supplementation on the antibody titer to heat-shock protein (HSP) 70, hsCRP and spirometry test in patients with mild and moderate persistent allergic asthma: A triple-blind, randomized placebo-controlled trial.

BACKGROUND: Asthma is a heterogeneous disease, which usually associated with chronic airway inflammation. The anti-heat shock protein (anti-HSP) 70 is a novel risk factor for asthma. The aim of the present study was to survey the effect of saffron supplementation on anti-HSP70, high-sensitivity C-reactive protein (hs-CRP) and spirometry test in patients with allergic asthma. BASIC PROCEDURES: In this clinical trial, patients (N = 80, 32 women and 48 men, 18-65 years old) with mild and moderate allergic asthma were randomized into two groups: a group of patients who received two capsules of saffron (100 mg/d) and a control group who received two capsules of placebo for 8 weeks. Anti-HSP70, hs-CRP and spirometry test were determined in patients before (week 0) and after (week 8) intervention. SPSS software (version 16.0; Inc, Chicago, IL) was used for data analysis. MAIN FINDINGS: Saffron in comparison with placebo significantly reduced the hs-CRP (p < 0.001) and anti-HSP70 (p < 0.001) concentrations. In spirometry test, forced expiratory volume in first second(FEV1), forced vital capacity (FVC), FEV1/FVC ratio and forced expiratory flow 25-75%.(FEF 25-75) increased significantly in saffron in comparison to placebo group (p < 0.05). PRINCIPAL CONCLUSIONS: Results of the present study suggested that saffron supplementation in patients with allergic asthma decreased significantly anti-HSPs 70 and hs-CRP and also improved some spirometry test factors.

Biological effects of Er:YAG laser irradiation on the proliferation of primary human gingival fibroblasts.

We investigated the biological effects of Er:YAG laser (2940-nm; DELight, HOYA ConBio, Fremont, California) irradiation at fluences of 3.6, 4.2, 4.9, 6.3, 8.1 or 9.7 J cm-2 at 20 or 30 Hz for 20 or 30 seconds on primary human gingival fibroblasts (HGFs). Irradiation at 6.3 J cm-2 promoted maximal cell proliferation, determined by WST-8 assay and crystal violet staining, but was accompanied by lactate dehydrogenase release, on day 3 post-irradiation. Elevation of ATP level, Ki67 staining, and cyclin-A2 mRNA expression confirmed that Er:YAG affected the cell cycle and increased the number of proliferating cells. Transmission electron microscopy showed alterations of mitochondria and ribosomal endoplasmic reticulum (ER) at 3 hours post-irradiation at 6.3 J cm-2 , and the changes subsided after 24 hours, suggesting transient cellular injury. Microarray analysis revealed up-regulation of 21 genes involved in heat-related biological responses and ER-associated degradation. The mRNA expression of heat shock protein 70 family was increased, as validated by Real-time PCR. Surface temperature measurement confirmed that 6.3 J cm-2 generated heat (40.9°C post-irradiation). Treatment with 40°C-warmed medium increased proliferation. Laser-induced proliferation was suppressed by inhibition of thermosensory transient receptor potential channels. Thus, despite causing transient cellular damage, Er:YAG laser irradiation at 6.3 J cm-2 strongly potentiated HGF proliferation via photo-thermal stress, suggesting potential wound-healing benefit.

Role of HSP70 in ozone preconditioning-induced reduction of hepatic ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To evaluate the role of heat shock protein 70 (HSP70) in ozone preconditioning-induced reduction of hepatic ischemia-reperfusion (I/R) injury in rats.Methods Twenty-four pathogen-free healthy male Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were divided into 4 groups (n=6 each) using a random number table:sham operation group (group S),group I/R,ozone preconditioning group (group OP) and HSP70 inhibitor quercetin group (group Q).Hepatic I/R injury was induced by clamping the branches of hepatic artery supplying left and middle lobes of the liver,portal vein and bile vessel for 45 min,followed by 2 h of reperfusion.In group OP,the mixture of ozone and oxygen was intraperitoneally injected at the concentration of ozone 50 μg/ml and bolus dose 1 mg/kg once a day for 5 consecutive days before operation.In group Q,quercetin 7 mg/kg was intraperitoneally injected at 1 h before injecting the mixture of ozone and oxygen.Blood samples from the abdominal aorta were collected at 2 h of reperfusion for determination of serum alanine transaminase (ALT) and aspartate transaminase (AST) activities and concentrations of interleukin-1beta (IL-1β),IL-6 and tumor necrosis factor-alpha (TNF-α) in serum (by enzyme-linked immunosorbent assay).The animals were sacrificed after blood sampling,and hepatic tissue specimens were collected for detection of activated caspase-3 expression (by Western blot) and cell apoptosis (by TUNEL).Apoptotic index (AI) was calculated.Results Compared with group S,the serum ALT and AST activities,concentrations of IL-1β,IL-6 and TNF-α in serum and AI were significantly increased,and the expression of activated caspase-3 was up-regulated in group I/R (P＜0.05 or 0.01).Compared with group I/R,the serum ALT and AST activities,concentrations of IL-1β,IL-6 and TNF-α in serum and AI were significantly decreased,and the expression of activated caspase-3 was downregulated in group OP (P＜0.05 or 0.01).Compared with group OP,the serum ALT and AST activities,concentrations of IL-1β,IL-6 and TNF-α in serum and AI were significantly increased,and the expression of activated caspase-3 was up-regulated in group Q (P＜0.05 or 0.01).Conclusion The mechanism by which ozone preconditioning reduces hepatic I/R injury is related to activating HSP70 and inhibiting inflammatory responses and cell apoptosis in rats.

Effect of trimetazidine on expression of HSP70 and Mn-SOD after myocardial ischemia/reperfusion in rats / 中华老年心脑血管病杂志

Objective To study the protective effect of trimetazidine pretreatment on myocardial I/R in a rat I/R model and its mechanism.Methods One hundred and twenty SD rats were divided into control group,I/R group,HSP70 inhibitor group,trimetazidine treatment group and combined treatment group (24 in each group).The rats in each group were further divided into 30 min reperfusion group,4 h reperfusion group and 8 h reperfusion group (8 in each group).Their HSP70-positive myocardial cells were calculated,their serum CK,CK-MB,MDA,SOD levels and their myocardial infarction size were measured.Results The serum CK and CK-MB levels were significantly higher in I/R group,HSP70 inhibitor group,trimetazidine treatment group and combined treatment group than in control group and were significantly lower in trimetazidine treatment group than in combined treatment group and I/R group after reperfusion for 4 and 8 h (P＜0.01).The serum levels of HSP70 and SOD were significantly higher,the MDA levels were significantly lower in trimetazidine treatment group than in combined treatment group and I/R group after reperfusion for 30 min,4 and 8 h (P＜0.01).The infraction size was significantly smaller in trimetazidine treatment group than in combined treatment group and I/R group after reperfusion for 4 and 8 h (P＜0.05).Conclusion Trimetazidine can increase the serum HSP70 level in myocardial cells,reduce the infarction size,and protect the myocardium against I/R in rats.

Research advances in immune enhancement by heat shock protein 70 induced by local ablation for liver cancer / 临床肝胆病杂志

Liver cancer still threatens human health nowadays, so it is urgent to explore new minimally invasive methods for the treatment of liver cancer. This article reviews the general status of liver cancer in China and describes local ablation and immunotherapy for liver cancer. This article also introduces the basic principle of nanosecond pulse in the treatment of liver cancer and the mechanism of action of heat shock protein 70 (HSP70) in antitumor immunity and points out that as an exogenous electrical stimulation, nanosecond pulse can increase the expression of HSP70 in the body to strengthen the body′s anti-tumor immunity. This article perfects the immunological mechanism of nanosecond pulse in the treatment of liver cancer and provides new thoughts for immunotherapy for liver cancer.