Up-regulation of Hsp72 and keratin16 mediates wound healing in streptozotocin diabetic rats

BACKGROUND: Impaired wound healing is a complication of diabetes and a serious problem in clinical practice. We previously found that whey protein (WP) was able to regulate wound healing normally in streptozotocin (STZ)-dia-betic models. This subsequent study was designed to assess the effect of WP on heat shock protein-72 (Hsp72) and keratin16 (Krt16) expression during wound healing in diabetic rats. METHODS: WP at a dosage of 100 mg/kg of body weight was orally administered daily to wounded normal and STZ-diabetic rats for 8 days. RESULTS: At day 4, the WP-treated diabetic wound was significantly reduced compared to that in the corresponding control. Diabetic wounded rats developed severe inflammatory infiltration and moderate capillary dilatation and regeneration. Treated rats had mild necrotic formation, moderate infiltration, moderate to severe capillary dilatation and regeneration, in addition to moderate epidermal formation. Hsp72 and Krt16 densities showed low and dense activity in diabetic wounded and diabetic wounded treated groups, respectively. At day 8, WP-treatment of diabetic wounded animals revealed great amelioration with complete recovery and closure of the wound. Reactivity of Hsp72 and Krt16 was reversed, showing dense and low, or medium and low, activity in the diabetic wounded and diabetic wounded treated groups, respectively. Hsp72 expression in the pancreas was found to show dense reactivity with WP-treated diabetic wound rats. CONCLUSION: This data provides evidence for the potential impact of WP in the up-regulation of Hsp72 and Krt16 in T1D, resulting in an improved wound healing process in diabetic models.

Effect of Heat Shock Protein 72 Expression on Etoposide-induced Cell Death of Rat Retinal Ganglion Cells

PURPOSE: To assess whether the expression of heat shock protein 72 (Hsp72) protects rat retinal ganglion cells (RGC-5) from apoptotic cell death. METHODS: Hsp72 expression in RGC-5 cells transduced with replication-deficient recombinant adenovirus was analyzed by Western blot analysis and immunofluorescence. The effect of Hsp72 expression on etoposide-induced apoptotic cell death was examined by microscopic analysis and confirmed by cell proliferation assay. RESULTS: Western blot analysis and immunofluorescence clearly showed adenovirus-mediated Hsp72 expression in RGC-5 cells. Treatment with etoposide resulted in the death of a proportion of the cells by apoptosis. However, this apoptotic cell death was significantly reduced in cells expressing Hsp72, with the reduction in cell death correlating to the level of Hsp72 expression. CONCLUSIONS: Over-expression of Hsp72 alone is sufficient to rescue neuronal cells from apoptotic cell death, suggesting that fine-tuning its expression may be an effective neuroprotective approach in retinal degenerative disease.

The effect of Hsp72 on IL-6, IL-8 expression and activation of NF-kappaB in synoviocytes of rheumatoid arthritis / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of heat shock protein 72 (Hsp72) on the expression of IL-6 and IL-8 and activation of NF-kappaB in synoviocytes from patients suffered from rheumatoid arthritis (RA).</p><p><b>METHODS</b>IL6 and IL8 concentrations in culture supernatants were measured using enzyme-linked immunosorbent assays (ELISA). Nuclear translocation of NF-kappaB and degradation of the inhibitory protein IkappaBalpha were examined using immunohistochemistry and Western blot.</p><p><b>RESULTS</b>Hsp72 down-regulated IL-6 and IL-8 production in RA synoviocytes induced by tumor necrosis factor-alpha (TNF-alpha). Hsp72 inhibited nuclear translocation of NF-kappaB and degradation of IkappaBalpha induced by TNF-alpha.</p><p><b>CONCLUSION</b>Hsp72 has an anti-inflammatory effect on RA by down-regulation of IL-6 and IL-8 in synoviocytes, which is mediated through inhibiting the activation of NF-KalphaB signal pathways.</p>

[Acute treadmill exercise increases heat shock protein 72 and total antioxidant capacity level in the streptozotocin- induced diabetic rat Hippocampus]

The purpose of this study was to investigate the effects of acute treadmill exercise on Hsp72 and TAC in the hippocampus of diabetic rats. Forty male rats, weight 165 +/- 1 g, were assigned randomly into 4 groups: Diabetic control group, and 3 diabetic+acute training groups, including groups 1,2 and 3, the animals of which were sacrificed 30 minutes, 4 hours and 24 hours postexercise respectively. Diabetes was induced by injecting streptozotocin [STZ; 0/5 mg/kg dissolved in 0.1 M citrate buffer, pH 4.5] into the abdominal cavity. Rats were subjected to treadmill exercise, [treadmill speed 10 m/min, gradually increased to 18 m/min, grade 0%, duration exercise 60 min]. Data were analysed with one-way Anova and tukey post hoc, P

Efeitos do treinamento em natação abaixo do limiar de lactato sobre a expressão de proteínas de estresse (Hsp72) no miocárdio de ratos / Effects of swimming training below lactate threshold on the expression of stress proteins (hsp72) in the myocardium of rats

Os objetivos deste estudo foram testar se programas de natação com intensidades abaixo do limiar de lactato induzem a expressão de proteínas de estresse (Hsp72) no miocárdio de ratos e se a indução da expressão de Hsp72 é distinta nos ventrículos direito e esquerdo. Ratos Wistar foram alocados randomicamente em três grupos: controle (C, n = 8); natação sem sobrecarga (NSS, n = 8); e natação com sobrecarga - 3% do peso corporal (NCS, n = 8). Animais NSS e NCS nadaram 30 minutos/dia, cinco dias/semana, durante sete semanas. Após eutanásia, o coração foi removido, pesado e foram coletados fragmentos dos ventrículos direito (VD) e esquerdo (VE) para análise dos níveis de Hsp72. O peso relativo do coração não foi diferente (p=0,68) entre os grupos (C=4,52 ± 0,87; NSS= 4,54 ± 0,79; NCS=4,72 ± 0,16 mg/g). Os níveis de Hsp72 foram maiores no VE do grupo NCS do que no C (396,29 ± 11,91 vs. 321,04 ± 9,65 unidade arbitrária, respectivamente; p = 0,0006). Hsp72 no VE do grupo NCS foram maiores que no NSS (396,29 ± 11,91 vs. 339,43 ± 10,21 unidade arbitrária, respectivamente; p = 0,004). Não houve diferença de Hsp72 no VD entre os grupos (C=320,02 ± 10,35; NSS=321,53 ± 24,8; NCS=353,08 ± 23,44 unidade arbitrária; p = 0,47). Não houve diferença na expressão de Hsp72 entre VD e VE nos grupos (C, p=0,94; NSS, p=0,52; NCS, p=0,11). Concluiu-se que o programa de natação com intensidade abaixo do limiar de lactato (3% do peso corporal) induz a expressão de HSP 72 no miocárdio de ratos, especialmente no ventrículo esquerdo.

We tested whether swimming training regimes of different intensities bellow the anaerobic threshold induce expression of stress protein (Hsp72) in rat’s myocardium; and if the expression of Hsp72 differs between left and right ventricles. Wistar rats (6 weeks of age, body weight of ~ 261 g) were allocated into three groups: control (C, n = 8); unloaded swimming (NSS, n = 8); and loaded swimming - 3% body weight (NCS, n = 8). Animals from NSS and NCS swam 30 min/day, 5 days/week, during 7 days. At sacrifice the heart was removed and weighed. Fragments of right (RV) and left ventricles (LV) were harvested and the levels of Hsp72 determined. The heart weight to body weight ratio did not differ among C, NSS and NCS groups (4.52 ± 0.87; 4.54 ± 0.79; 4.72 ± 0.16 mg/g, respectively). The LV levels of Hsp72 were higher in NCS group than in C (396.29 ± 11.91 vs. 321.04 ± 9.65 arbitrary unit, respectively, p = 0.0006). The LV levels of Hsp72 were higher in NCS than in NSS (396.29 ± 11.91 vs. 339.43 ± 10.21 arbitrary unit, respectively; p = 0.004). There was no difference in RV levels of Hsp72 among groups (C=320.02 ± 10.35; NSS=321.53 ± 24.80; NCS=353.08 ± 23.44 arbitrary unit; p = 0.47). There was no difference in Hsp72 levels between RV and LV among groups (C, p=0.94; NSS, P=0.52; NCS, p=0.11). It was concluded that the swimming training program with intensity bellow the anaerobic threshold (3% body weight) induces expression of Hsp72 in the rat myocardium, especially in the left ventricle.

Effects of graded hypothermia on hypoxic-ischemic brain damage in the neonatal rat / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effect of graded hypothermia on neuropathologic alterations of neonatal rat brain after exposed to hypoxic-ischemic insult at 37°C, 33°C, 31°C, and 28°C, respectively, and to observe the effect of hypothermia on 72-kDa heat shock protein (HSP72) expression after hypoxic-ischemic insult.</p><p><b>METHODS</b>Seven days old Wistar rats were subjected to unilateral common carotid artery ligation followed by exposure to hypoxia in 8% oxygen for 2 hours at 37°C, 33°C, 31°C, and 28°C, respectively. The brain temperature was monitored indirectly by inserting a mini-thermocouple probe into the temporal muscle during hypoxia. After hypoxia-ischemia their mortality was assessed. Neuronal damage was assessed with HE staining 72 hours after hypoxia. HSP72 expression at 0.5, 24, and 72 hours of recovery was immunohistochemically assessed using a monoclonal antibody to HSP72.</p><p><b>RESULTS</b>Hypoxia-ischemia caused 10.5% (2/19) of mortality in rat of 37°C group, but no death occurred in 33°C, 31°C or 28°C groups. HE staining showed neuropathologic damage was extensive in rats exposed to hypoxia-ischemia at 37°C (more than 80.0%). The incidence of severe brain damage was significantly decreased in 33°C (53.3%) and 31°C groups (44.4%), and no histologic injury was seen in the 28°C group of rats. Expression of HSP72 was manifest and persistent in the rat brain of 37°C group, but minimum in the rat brain of 28°C group.</p><p><b>CONCLUSION</b>Mild and moderate hypothermia might prevent cerebral visible neuropathologic damage associated with hypoxic-ischemic injury by decreasing stress response.</p>

Physiological stress responses in the warm-water fish matrinxã (Brycon amazonicus ) subjected to a sudden cold shock / Respostas fisiológicas ao estresse do peixe de águas tépidas matrinxã (Brycon amazonicus ) submetido à queda brusca de temperatura

The present work evaluated several aspects of the generalized stress response [endocrine (cortisol), metabolic (glucose), hematologic (hematocrit and hemoglobin) and cellular (HSP70)] in the Amazonian warm-water fish matrinxã (Brycon amazonicus ) subjected to an acute cold shock. This species farming has been done in South America, and growth and feed conversion rates have been interesting. However, in subtropical areas of Brazil, where the water temperature can rapidly change, high rates of matrinxã mortality have been associated with abrupt decrease in the water temperature. Thus, we subjected matrinxã to a sudden cold shock by transferring the fish directly to tanks in which the water temperature was 10ºC below the initial conditions (cold shock from 28ºC to 18ºC). After 1h the fish were returned to the original tanks (28ºC). The handling associated with tank transfer was also imposed on control groups (not exposed to cold shock). While exposure to cold shock did not alter the measured physiological conditions within 1h, fish returned to the ambient condition (water at 28º C) significantly increased plasma cortisol and glucose levels. Exposure to cold shock and return to the warm water did not affect HSP70 levels. The increased plasma cortisol and glucose levels after returning the fish to warm water suggest that matrinxã requires cortisol and glucose for adaptation to increased temperature.

O presente trabalho avaliou as principais respostas fisiológicas e celulares [endócrino (cortisol), metabólico (glicose), hematológico (hematócrito and hemoglobina) e celular (HSP70)] ao estresse de um peixe de águas tépidas, o matrinxã (Brycon amazonicus ), quando submetido a um choque térmico frio abrupto. Essa espécie vem sendo amplamente cultivada na América do Sul por apresentar excelentes índices zootécnicos de crescimento e conversão alimentar. Entretanto, os produtores rurais encontram limitações no manejo do matrinxã, quando criado em regiões mais frias que sua região de origem, a Bacia Amazônica. Assim, o matrinxã foi submetido a um choque frio através da transferência direta dos peixes para tanques com água fria a 18ºC. Após 1h, esses peixes retornaram a suas caixas de origem a 28ºC. O manuseio de peixes necessário para conduzir o choque térmico experimental foi também imposto aos grupos controle, sendo, entretanto, evitada a água fria. O matrinxã demonstrou claros sinais de estresse fisiológico durante os procedimentos experimentais. Porém, essas respostas não foram associadas ao choque frio, mas sim ao choque quente por ocasião da volta dos peixes para as caixas de origem. As respostas primárias e secundárias de estresse foram evidentes através das análises plasmáticas de cortisol e glicose. Já o hematócrito, a hemoglobina e as expressões da proteína de estresse, HSP70, não foram afetadas. Nossos resultados sustentam que o matrinxã falhou em responder ao choque térmico frio, mas não ao choque térmico quente, que é um estressor evidentemente associado à origem natural dessa espécie de águas de elevadas temperaturas.

Nitric oxide induces heat shock protein 72 production and delayed protection against myocardial ischemia in rabbits via activating protein kinase C / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Nitric oxide (NO) is a biologically active molecule which has been reported to protect the heart against ischemia and reperfusion injury in different species. This study aimed to test the hypothesis that nitric oxide may induce the expression of heat shock protein 72 (HSP72) which may protect the heart against ischemia.</p><p><b>METHODS</b>Rabbits were given intravenous saline or S-nitroso-N-acetylpenicillamine (SNAP), a nitric oxide donor, or Zaprinast, an inhibitor of cyclic guanosine monophosphate (GMP)-phosphodiesterase, which may increase myocardial cyclic GMP content. Twenty-four hours later, the rabbits were either sampled to measure HSP72, or induced with a 30-minute coronary occlusion followed by a 120-minute reperfusion, and then the infarct size was measured. Meanwhile, chelerythrine (CHE, an inhibitor of protein kinase C) was given intravenously 5 minutes before SNAP injection and the effect on HSP72 expression and infarct size was determined.</p><p><b>RESULTS</b>Twenty-four hours after pretreatment, immunoblotting showed HSP72 expression increased in the SNAP group compared with control groups, and this was blocked by CHE. Myocardial infarct size in the SNAP group was smaller than that of the control group ((32.4 +/- 5.8)% vs (51.1 +/- 4.7)%, P < 0.05). Pretreated with CHE abolished the infarct size-limiting effect of SNAP ((46.0 +/- 5.1)%). Pretreatment with Zaprinast neither induced HSP72 expression nor reduced infarct size ((55.4 +/- 5.4)%).</p><p><b>CONCLUSION</b>NO induced HSP72 expression and a delayed protection to the heart via the activities of protein kinase C by a cyclic GMP-independent pathway.</p>

Influence of exercise at high temperature on blood biochemical indexes and HSP72 expression in adult males / 华中科技大学学报（医学）（英德文版）

The influence of exercise at high temperature on adult males' routine blood indexes and biochemical indexes and the expression of HSP72 in peripheral blood lymphocytes (PBLs) was studied in order to provide theoretical ground for health supervision of adults receiving exercise at high temperature. 180 adult males were selected and divided into exercise group and control group, in which the exercise group was subdivided into subgroup 1 and subgroup 2 receiving exercise at high temperature in the afternoon and in the morning, respectively. Peripheral venous blood was phlebotomized before and after the exercise to examine routine blood indexes and blood biochemical indexes. The expression levels of HSP72 in PBLs were detected by flow cytometry. The results showed that the routine blood indexes and biochemical indexes in each group were within the range of normal values of male adults. There was no significant difference between each exercise group and control group in indexes before exercise. After exercise, the expression levels of HSP72 in PBLs in exercise groups were higher than those before exercise, and HSP72 expression levels in subgroup 1 were obviously higher than those in subgroup 2 and control group. The contents of ALT, urea, Na+, Cl-, Ca2+ and K+ in subgroups 1 and 2 were lower than those in control group, but CK level was higher than in control group (P<0.05). The contents of Na+ and Cl- in subgroup 1 were relatively lower than those in subgroup 2 (P<0.05). It was concluded that while receiving exercise at high temperature, adult males' HSP72 levels in PBLs could be increased and the biochemical indexes changed. Attention should be paid to health supervision to avoid obvious body injuries at high temperature.

Heat preconditioning reduces subsequent ischemia reperfusion injury in rat kidney: a possible role for HSP-72

Improving the ability of the kidney to tolerate ischemic injury has important implication in renal transplantation. On the other hand, thermo tolerance describes the process in which hyperthermia induces a transient resistance of the stressed cells to subsequent episodes of oxidative stress. The current study was performed to evaluate the beneficial effect of heat preconditioning induced HSP-72 formation on renal ischemia reperfusion [I/R] induced damage. Four groups of rats [n=20/group] were included: Control sham-operated group [group I], heat-preconditioned sham-operated group [group II], I/R injury group [group III] and heat pre-conditioned I/R injury group [group IV]. Heat-preconditioning was induced 24h prior to sham operation and or I/R injury by increasing the core body temperature to [41 +/- 0.5°C] for 20min. The rat kidneys were subjected to ischemia by 20min of bilateral renal artery occlusion followed by reperfusion for 24 and 48h. After 24 and 48h of reperfusion, serum urea, creatinine, 24h urine out put and albumin content as well as the renal HSP-72 gene expression and MDA level were measured. Also light microscopic examination of renal tissue specimens was performed. It was found that group IV had a significant increase in renal HSP-72 gene expression compared to group III [898.36 +/- 107.82 versus 572.88 +/- 47.08 micro g/g tissue], associated with a significant improvement of its renal functions including serum urea, creatinine and 24h urine volume out put. Also there was a reduction in renal tissue injury detected by a significant decrease in urine albumin content, a significant decrease in renal MDA level and improvement in specimen microscopic picture compared to group III. The increase in HSP-72 expression and its renoprotective effect were significantly greater 24h after I/R than after 48h. Thus it can be concluded that upregulation of HSP-72 after heat preconditioning has a renal beneficial effect and can be a target for protection of renal functions during I/R injury

Mechanisms of HSP72 release.

Currently two mechanisms are recognized by which heat shock proteins (HSP) are released from cells; a passive release mechanism, including necrotic cell death, severe blunt trauma, surgery and following infection with lytic viruses, and an active release mechanism which involves the non classical protein release pathway. HSPs are released both as free HSP and within exosomes. This review covers recent findings on the mechanism by which stress induces the release of HSP72 into the circulation and the biological significance of circulating HSP72 to host defense against disease.

Relationship between heat shock protein 72 and DNA genetic damage in peripheral blood lymphocytes of coke oven workers / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the relationship between heat shock protein 72 (Hsp72) and DNA genetic damage in peripheral blood lymphocytes of coke oven workers and the role of Hsp72 in protection of cells from genetic damage induced by coke oven emissions.</p><p><b>METHODS</b>Two hundred and sixty-seven coke oven workers and thirty controls without occupational PAHs exposure were investigated. Benzo[a]pyrene concentrations in the ambient air individually collected were assayed by high performance liquid chromatography (HPLC). Western Blot was used to measure Hsp72 levels and Comet assay was used to evaluate DNA damage degree. Personal information was collected by questionnaire.</p><p><b>RESULTS</b>The Hsp72 level (G+/-S(G)) and olive comet tail moment (G+/-S(G) of peripheral blood lymphocytes in high-exposure workers (1.24 +/- 0.42 and 4.49 +/- 1.24) were significantly higher than those in low-exposure workers (1.01 +/- 0.35 and 2.99 +/- 1.10, P < 0.05) and control (0.85 +/- 0.34 and 2.40 +/- 1.00, P < 0.05) respectively. The Hsp72 median level of all subjects was used as the limit to divide subjects into high Hsp72 level group and low Hsp72 level group. The rate with high Hsp72 level was 36.7%, 43.1% and 58.3% in control, low exposure and high exposure workers respectively and had a rising tendency following exposure level (P = 0.003). In high Hsp72 level group Hsp72 level in high exposure workers was significantly higher than that in control (P < 0.05), and there was a rising tendency along with the increase of exposed levels. But the olive comet tail moment had no significant difference among three exposed groups (P > 0.05). In low Hsp72 level group there no difference among three exposed groups about Hsp72 levels. The olive comet tail moment in high exposure workers was significantly higher than that in low exposure workers and control (P < 0.01) and high exposure workers in Hsp72 positive group and there was a rising tendency along with the increase of exposed levels. Hsp72 levels had strong negative correlation with the olive comet tail moment (r = -0.503, P < 0.01) in high exposure workers.</p><p><b>CONCLUSION</b>The coke oven emissions can induce hsp72 expression. Hsp72 play a role of protecting cells from DNA damage induced by coke oven emissions.</p>

Localization of Heat Shock Protein 72 in Potassium-deprivated Rat Kidney / 대한해부학회지

Heat shock protein 72 (HSP72) appears to play an important role in cell survival in the hypertonic conditions of the renal medulla. The purpose of this study was to examine the effect of potassium deprivation on renal HSP72 expression. Male Sprague-Dawley rats were fed potassium deficient diet for 2 weeks. Kidney tissues were preserved by in vivo perfusion with paraformaldehyde-lysine-periodate (PLP) and processed for Western blot analysis and immunocytochemistry. Serum potassium concentration and urine osmolality decreased in potassium deprivated animals. In control kidneys, HSP72 immunostaining was observed mainly in the inner medulla in almost all cells including the inner medullary collecting duct and papillary surface epithelial cells. In potassium deprivated kidneys, HSP72 expression decreased dramatically in the inner medulla. However, strong HSP72 immunostaining remained in some inner medullary collecting duct and papillary surface epithelial cell. These results demonstrated that potassium deprivation induced down regulation of HSP72 in the renal medulla, at least in part, through cell-specific manner.

Increased Expression of Heat Shock Protein 72 Protects Renal Proximal Tubular Cells from Gentamicin-induced Injury

The nephrotoxicity of gentamicin (GM) has been widely recognized. Heat shock protein 72 (HSP72) has been reported to be a cytoprotectant. However, its cytoprotective effect against GM induced kidney injury has not yet been studied. In this study, we investigated the cytoprotective effect of HSP72 on GM-induced nephrotoxicity in vitro. Human Kidney tubular cell line, HK-2 cells were divided into four groups: control group, GM group (cells incubated with GM only), heat shock (HS) group (cells incubated at 43 degrees C for 30 min), and GM plus HS group, respectively. Lactate dehydrogenanse (LDH) release increased time-dependently from 24 hr to 96 hr compared to the data of cells treated with GM only. Results of NAG activities, superoxide dismutase (SOD) activities and malondialdehyde (MDA) content were similar to that of the LDH release. The amount of HSP72 positive cells increased significartly at 72 hr after cells were treated with GM only. Both HSP72 protein and gene expression increased significantly at 72 hr when cells were treated with GM. On the other hand, HS induced HSP72 expression markedly. Pretreatment of HS inhibited HK-2 cells from GM-induced injury. It could reduce LDH release and NAG activity. HS also increased SOD activity, and decreased MDA content when cells were damaged by GM. These findings suggested that HS may protect kidney cells from GM-induced injury. Pre-induction of HSP72 may provide therapeutic strategies for nephrotoxicity induced by GM.

The protective effect of heat shock protein 72 by Doxorubicin in cold ischemia-reperfusion injury of the rat liver / 中华外科杂志

<p><b>OBJECTIVE</b>To observe induction of heat shock reaction by pretreatment of Doxorubicin (DXR) in long-term cold preservation-reperfusion injury of the rat liver.</p><p><b>METHODS</b>The rats were administered intravenously by DXR at a dose of 1 mg/kg body weight in DXR group and by saline in control group. After 48 hours, the rat liver was perfused by using cold University of Wisconsin (UW) solutions and was preserved in UW solution at 4 degrees C for 48 hours. Recipient liver was perfused for 1 and 3 hours after orthotopic liver transplantation. Tumor necrosis factor-alpha (TNF-alpha) mRNA, cytokine-induced neutrophil chemoattractant (CINC) mRNA, macrophage inflammatory protein (MIP-2) mRNA was measured by RT-PCR and heat shock protein 72 (HSP72), nuclear factor-kappaB (NF-kappaB) by Western blot. The serum levels of TNF-alpha, CINC, MIP-2 by ELISA and AST were measured. The survival rate of 7 days was observed.</p><p><b>RESULTS</b>The expression of TNF-alpha mRNA, CINC mRNA and MIP-2 mRNA was stronger in control group than in DXR group. HSP72 was expressed in SA group but not in control group and oppositely NF-kappaB was expressed in control group but not in DXR group. Serum AST, TNF-alpha, CINC and MIP-2 concentrations were significantly lower in DXR group than in control group (P < 0.05). The survival rate of 7 days was significantly higher in DXR group than in control group (50% vs. 0%, P < 0.05).</p><p><b>CONCLUSIONS</b>These data suggested that long-term cold ischemia-reperfusion injury was attenuated in liver graft with pretreatment of DXR. The induction of HSP72 may offer protection to hepatocytes by restraining the activation of NF-kappaB and inflammation.</p>

Relationship between dust exposure and chronic obstructive pulmonary diseases and heat shock protein 72 and 73 in lymphocytes among coal miners / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To assess the expression of Hsp72 and Hsp73 in chronic obstructive pulmonary disease (COPD) and to evaluate their roles in damage from coal dust exposure.</p><p><b>METHODS</b>A case control study of 50 coal miners suffering from COPD and 50 healthy coal miners were selected from one coal mine. The levels of Hsp72 and Hsp73 in peripheral blood lymphocytes were determined by flow cytometry for all subjects.</p><p><b>RESULTS</b>(1) The expression of basic Hsp72 of peripheral blood lymphocytes for patients and controls was not different from that inducible expressed Hsp72 by 42 degrees C heat stress or by BPDE exposure. (2) The expression of Hsp72 in COPD patients (17.7 +/- 4.9) was significantly lower than that in healthy coal miners (22.6 +/- 10.0) (P < 0.01). On the other hand, the expression of Hsp73 in COPD patients (33.5 +/- 11.7) was higher than that in healthy coal miners (19.6 +/- 5.9) (P < 0.01). (3) A-positive relationship between the expression of Hsp72 and cumulative inhaling coal dust exposure was observed. No relationship was found between Hsp73 and cumulative inhaling coal dust exposure.</p><p><b>CONCLUSION</b>The decreased expressions of Hsp72 in peripheral blood lymphocytes of COPD coal miners.</p>

Effect of HSP72 on acute injury of neonatal cardiomyocytes induced by oxidative stress / 中南大学学报(医学版)

OBJECTIVE@#To explore the protective effect of HSP72 on the acute injury of cardiomyocyte induced by oxidative stress.@*METHODS@#Cardiomyocytes of neonatal rats treated with heat shock (42 degrees C, 30 min, recovery for 6 h) to induce the expression of HSP72 and HSP72 antisense oligonucleotide was transformed to block the expression of HSP72. 0.5 mmol/L (final concentration) H2O2 was added into the culture medium to mimic oxidative stress, and to induce the acute injury of neonatal cardiomyocytes. The release of LDH and the total protein synthesis were applied to evaluate the injury of cardiomyocyte of neonatal rats.@*RESULTS@#Oxidative stress could significantly increase the release of LDH, and inhibit the total protein synthesis. By inducing the expression of HSPs, heat shock pretreatment significantly reduced the release of LDH and relieved the oxidative stress-mediated inhibition of total protein synthesis. Moreover, HSP7-2 anti-sense oligonucleotide could remarkably block the protective effect of heat shock pretreatment on the cellular injuries induced by H2O2.@*CONCLUSION@#HSP72 plays a most important role in the acute injury of cardiomyocyte mediated by oxidative stress.

Vitamin C supplement modulates heat shock protein 72 gene expression in skeletal muscle of rats with type 2 diabetes

Oxidative stress has been ascribed a role in the pathogenesis of diabetes and its complications and stress proteins have been shown to protect organisms in vitro and in vivo against oxidative stress. In this study we examined the HSP72 gene expression in skeletal muscle of type 2 diabetic rats induced by oral fructose administration [66% of total caloric intake] compared to control and the possibility to increase its level by oral administration of vitamin C [100mg/kg/day for 8 weeks]. The amount of HSP72 m RNA in muscles of diabetic rats was lower than in control non-diabetic group [20.3 +/- 6.37 versus 40.52 +/- 7.49 micro g/g tissue] and its expression increased significantly by vitamin C administration [51.41 +/- 22.54 micro g/g tissue]. There was an insignificant increase in muscle insulin-stimulated glucose uptake after vitamin C administration in diabetic rats [2.59 +/- 0.66mg/g tissue versus 2 +/- 0.7mg/g tissue in diabetics not receiving vitamin C]. The plasma glucose level following vitamin C administration in the diabetic group, showed a significant negative correlation with the expression of HSP72. The concentration of malondialdehyde [MDA] as a measure of lipid peroxidation increased significantly in diabetics [2.019 +/- 0.53 micro mol/g tissue] compared to control group [0.926 +/- 0.19 micro mol/g tissue] and administration of vitamin C in diabetic rats decreased the concentration of MDA insignificantly [1.55 +/- 0.47 micro mol/g tissue] compared to the diabetic group not receiving vitamin C. In conclusion, the finding of decreased levels of HSP72 expression and decreased insulin-stimulated glucose uptake in skeletal muscle of type 2 diabetic rats raises the possibility that heat shock proteins may be involved in the pathogenesis of skeletal muscle insulin resistance in type 2 diabetics. Administration of vitamin C could be used in diabetics to increase heat shock protein HSP72 expression and to improve insulin resistance

Increased expression of 70 kD heat shock protein in cultured primary human keratinocytes induced by human papillomavirus 16 E6/E7 gene / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Heat shock protein 70 (HSP70) is expressed highly in epithelial tumours associated closely with human papillomavirus 16 (HPV16) infections. However, evidence about the direct relationship between HSP70 expression and HPVs infections are still lacking. In the present study, we examined the expression of HSP70 in keratinocytes introduced with HPV16 E6/E7 oncogenes.</p><p><b>METHODS</b>Stable transfected cells were established by transfection of the plasmids pLXSN16E6/E7 into cultured primary keratinocytes and subsequently selected by plasmid specific selection antibiotic (G418) at the required concentration. The expression of HSP70 in pLXSN16E6/E7 transfected keratinocytes was determined by Western blot. The correlation of HSP70 expression and E6/E7 transfection was further confirmed by doubly labelled immunofluorescent staining.</p><p><b>RESULTS</b>Compared to non-transfected keratinocytes, there was a significant trend for higher levels of HSP70 in pLXSN16E6/E7 transfected keratinocytes. Doubly labelled immunofluorescent staining experiment showed that the co-localization of HPV16 E6/E7 and HSP70 in transfected keratinocytes was observed and increased expression of HSP70 was strongly associated with the transfection of HPV16 E6/E7.</p><p><b>CONCLUSIONS</b>Our studies demonstrated increased levels of HSP70 proteins in keratinocytes stably transfected by HPV16 E6/E7 oncogenes. It suggests that the expression of HSP70 is modulated by HPV16 E6/E7 proteins, which may be involved in HPV16 E6/E7 induced immortalization.</p>

Nitric oxide opens second window of protection in ischemic preconditioning via induction of heat-shock protein 72 / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To examine the inhibition of nitric oxide (NO) synthesis during ischemic preconditioning (IP) upon the induction of heat-shock protein 72 (HSP72) and the size-limiting effect of the second window of protection on infarction.</p><p><b>METHODS</b>Rabbits were subjected to either 4 cycles of 5-min long coronary artery occlusion separated by 10 min of reperfusion, or a sham operation. During this procedure, we administered 10 mg/kg of N(G)-nitro-L-arginine methyl ester (L-NAME, an inhibitor of NO synthase) intravenously 5 min before IP followed by its continuous infusion (1.5 mg/kg/min). Twenty-four hours after IP or the sham operation, the hearts were rapidly excised for assay of HSP72 expression or were subjected to 30 min of coronary artery occlusion followed by 120 min of reperfusion, at which point infarct size (IS) was measured.</p><p><b>RESULTS</b>Twenty-four hours after IP or the sham operation, there was no difference in heart rate or mean arterial pressure between the groups. Immunoblotting revealed an increase in HSP72 protein levels in the IP group, which was blocked by L-NAME. IS in the IP rabbits was reduced compared with controls (29.8 +/- 3.7% vs. 50.8 +/- 4.3%, P < 0.01). IS in the IP rabbits was elevated as a result of L-NAME treatment (46.0 +/- 5.1%). Administration of L-arginine reversed the effects of L-NAME on the induction of HSP72 and IS (33.5 +/- 4.0%). The intravenous administration of S-nitroso-N-acetylpenicillamine (an NO donor, 15 microg/kg/min) over 20 min increased the induction of HSP72 and reduced IS (31.3 +/- 5.7%, P < 0.01 vs. control) 24 h later.</p><p><b>CONCLUSION</b>These findings suggest that NO may be involved in the induction of HSP72 and the opening of the second window of protection of IP.</p>