ABSTRACT
OBJECTIVE@#To investigate the anti-oxidative effect of ethyl pyruvate (EP) and taurine (TAU) on the quality of red blood cells stored at 4±2 ℃, hemolysis, energy metabolism and lipid peroxidation of the red blood cells in the preservation solution were studied at different intervals.@*METHODS@#At 4±2 ℃, the deleukocyte red blood cells were stored in the citrate-phosphate-dextrosesaline-adenine-1 (CPDA-1) preservation (control group), preservation solution with EP (EP-AS), and TAU (TAU-AS) for long-term preservation. The enzyme-linked immunoassay and automatic blood cell analyzer were used to detect hemolysis and erythrocyte parameters. Adenine nucleoside triphosphate (ATP), glycerol 2,3-diphosphate (2,3-DPG) and malondialdehyde (MDA) kits were used to test the ATP, 2,3-DPG and MDA concentration.@*RESULTS@#During the preservation, the rate of red blood cell hemolysis in EP-AS and TAU-AS groups were significantly lower than that in CPDA-1 group (P<0.01). The MCV of EP-AS group was increased with the preservation time (r=0.71), while the MCV of the TAU-AS group was significantly lower than that in the other two groups (P<0.05). The concentration of ATP and MDA in EP-AS and TAU-AS groups were significantly higher than that in CPDA-1 group at the 14th day (P<0.01). The concentrations of 2,3-DPG in the EP-AS and TAU-AS groups were significantly higher than that in the CPDA-1 group from the 7th day (P<0.01).@*CONCLUSION@#EP and TAU can significantly reduce the red blood cell hemolysis rate, inhibit the lipid peroxidation level of red blood cells, and improve the energy metabolism of red blood cells during storage. The mechanism of EP and TAU may be related to their antioxidation and membrane protection effect, so as to improve the red blood cell quality and extend the preservation time.
Subject(s)
Humans , 2,3-Diphosphoglycerate/metabolism , Adenine , Adenosine Triphosphate/metabolism , Blood Preservation , Citrates/pharmacology , Erythrocytes/metabolism , Glucose/pharmacology , Hemolysis , Pyruvates , Taurine/pharmacologyABSTRACT
Objective@#To examine the effects of ethyl pyruvate (EP) on mitochondrial dynamics and cell apoptosis in lipopolysaccharide (LPS)-induced human kidney-2 (HK-2) cells.@*Methods@#HK-2 cells were divided into three groups: HK-2 cells were challenged with LPS (800 μg/L) for 24 hours as LPS group, or LPS mixed with EP (0.25 mmol/L) for 24 hours as EP group. Cells were incubated with normal saline for 24 hours as control group. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and intracellular adenosine triphosphate (ATP) were detected by enzyme linked immunosorbent assay (ELISA). JC-1 staining and Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) assays were used to evaluate mitochondrial membrane potential and cell apoptosis, respectively. Western Blot was used to evaluate the protein expressions of mitochondrial dynamics, including death-associated protein kinase 2 (DAPK-2), mitofusin (Mfn-1 and Mfn-2), and apoptotic associated biomarkers, including caspase-3, caspase-9, Bcl-2, Bcl-xL, cytochrome C (Cyt C), and DNA repair enzyme poly ADP-ribose polymerase (PARP).@*Results@#Compared with the NC group, MDA, IL-6, TNF-α of LPS group were significantly increased, the expression of SOD, mitochondrial membrane potential and ATP level were significantly decreased, the expression of mitochondrial fission protein DAPK-2 was significantly increased, and mitochondrial fusion proteins Mfn-1 and Mfn-2 were significantly decreased, cell apoptosis and apoptotic protein caspase-3, caspase-9 and Cyt C were increased, and anti-apoptotic protein Bcl-2, Bcl-xL, PARP were significantly decreased. Compared with the LPS group, the oxidative activities and inflammatory factors above were inhibited in EP group [MDA (μmol/L): 12.35±2.21 vs. 45.95±1.76, SOD (kU/L): 54.68±1.42 vs. 40.73±1.60, IL-6 (ng/L): 67.87±2.61 vs. 338.92±20.91, TNF-α (ng/L): 19.23±1.80 vs. 180.69±6.51], mitochondrial membrane potential and ATP level were significantly increased [mitochondrial membrane potential: (99.43±0.25)% vs. (69.40±0.75)%, ATP (×106 RLU): 0.19±0.01 vs. 0.12±0.05], the expression of mitochondrial fission protein was significantly decreased (DAPK-2/β-actin: 0.03±0.01 vs. 0.61±0.02), mitochondrial fusion proteins were significantly increased (Mfn-1/β-actin: 0.43±0.04 vs. 0.17±0.01, Mfn-2/β-actin: 0.201±0.004 vs. 0.001±0.001), percentage of cell apoptosis was significantly decreased [(5.25±0.17)% vs. (34.42±0.64)%], the expressions of apoptotic proteins were significantly decreased (caspase-3/β-actin: 0.25±0.15 vs. 1.76±0.01, caspase-9/β-actin: 0.09±0.02 vs. 1.52±0.12, Cyt C/β-actin: 0.001±0.001 vs. 0.350±0.030), and the expressions of anti-apoptotic proteins and PARP were significantly increased (Bcl-2/β-actin: 0.500±0.010 vs. 0.009±0.004, Bcl-xL/β-actin: 0.550±0.010 vs. 0.009±0.001, PARP/β-actin: 0.94±0.01 vs. 0.16±0.13), with statistically significant differences (all P < 0.05).@*Conclusions@#There are enhanced mitochondrial fission and diminished mitochondrial fusion in LPS-induced HK-2 cells. EP can protect mitochondria functions by regulate mitochondrial dynamics, and reducethe apoptosis of LPS-induced HK-2 cells.
ABSTRACT
Objective To examine the effects of ethyl pyruvate (EP) on mitochondrial dynamics and cell apoptosis in lipopolysaccharide (LPS)-induced human kidney-2 (HK-2) cells. Methods HK-2 cells were divided into three groups: HK-2 cells were challenged with LPS (800 μg/L) for 24 hours as LPS group, or LPS mixed with EP (0.25 mmol/L) for 24 hours as EP group. Cells were incubated with normal saline for 24 hours as control group. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and intracellular adenosine triphosphate (ATP) were detected by enzyme linked immunosorbent assay (ELISA). JC-1 staining and Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) assays were used to evaluate mitochondrial membrane potential and cell apoptosis, respectively. Western Blot was used to evaluate the protein expressions of mitochondrial dynamics, including death-associated protein kinase 2 (DAPK-2), mitofusin (Mfn-1 and Mfn-2), and apoptotic associated biomarkers, including caspase-3, caspase-9, Bcl-2, Bcl-xL, cytochrome C (Cyt C), and DNA repair enzyme poly ADP-ribose polymerase (PARP). Results Compared with the NC group, MDA, IL-6, TNF-α of LPS group were significantly increased, the expression of SOD, mitochondrial membrane potential and ATP level were significantly decreased, the expression of mitochondrial fission protein DAPK-2 was significantly increased, and mitochondrial fusion proteins Mfn-1 and Mfn-2 were significantly decreased, cell apoptosis and apoptotic protein caspase-3, caspase-9 and Cyt C were increased, and anti-apoptotic protein Bcl-2, Bcl-xL, PARP were significantly decreased. Compared with the LPS group, the oxidative activities and inflammatory factors above were inhibited in EP group [MDA (μmol/L):12.35±2.21 vs. 45.95±1.76, SOD (kU/L): 54.68±1.42 vs. 40.73±1.60, IL-6 (ng/L): 67.87±2.61 vs. 338.92±20.91, TNF-α (ng/L): 19.23±1.80 vs. 180.69±6.51], mitochondrial membrane potential and ATP level were significantly increased [mitochondrial membrane potential: (99.43±0.25)％ vs. (69.40±0.75)％, ATP (×106 RLU): 0.19±0.01 vs. 0.12±0.05], the expression of mitochondrial fission protein was significantly decreased (DAPK-2/β-actin:0.03±0.01 vs. 0.61±0.02), mitochondrial fusion proteins were significantly increased (Mfn-1/β-actin: 0.43±0.04 vs. 0.17±0.01, Mfn-2/β-actin: 0.201±0.004 vs. 0.001±0.001), percentage of cell apoptosis was significantly decreased [(5.25±0.17)％ vs. (34.42±0.64)％], the expressions of apoptotic proteins were significantly decreased (caspase-3/β-actin: 0.25±0.15 vs. 1.76±0.01, caspase-9/β-actin: 0.09±0.02 vs. 1.52±0.12, Cyt C/β-actin: 0.001± 0.001 vs. 0.350±0.030), and the expressions of anti-apoptotic proteins and PARP were significantly increased (Bcl-2/β-actin: 0.500±0.010 vs. 0.009±0.004, Bcl-xL/β-actin: 0.550±0.010 vs. 0.009±0.001, PARP/β-actin:0.94±0.01 vs. 0.16±0.13), with statistically significant differences (all P < 0.05). Conclusions There are enhanced mitochondrial fission and diminished mitochondrial fusion in LPS-induced HK-2 cells. EP can protect mitochondria functions by regulate mitochondrial dynamics, and reducethe apoptosis of LPS-induced HK-2 cells.
ABSTRACT
Objective To study the roles of ethyl pyruvate ( EP ) in spinal cord edema after spinal cord injury ( SCI ) and in spinal cord astrocytic swelling after oxygen-glucose deprivation and reoxygenation ( OGD/R) in vitro in rats. Methods After SCI models were established in adult Sprague-Dawley ( SD ) rats, an intraperitoneal injection of EP was conducted to inhibit high mobility group box-1 ( HMGB1 ). Effects of EP on spinal cord edema, HMGB1 expression and astrocyte activation ( glial fibrillary acidic protein ( GFAP ) expression) in SCI rats were analyzed. Spinal cord astrocytes were cultured in post-natal SD rats and incubated under OGD/R procedure. Effects of EP on cell swelling, expression of HMGB1, aquaporin-4 ( AQP4 ) and toll-like receptor 4 ( TLR4 ) , and nuclear expression of nuclear factor-kappa B ( NF-κB ) in spinal cord astrocytes were observed. Results The water content in the spinal cord was increased significantly more at 1 d after SCI than at 12 h and 3 d ( P ＜0.05 ). Intraperitoneal injection of EP at 50 mg/kg reduced spinal cord water content, HMGB1 expression and astrocyte activation ( GFAP expression ) in SCI rats signif-icantly more than that at 25 mg/kg or 100 mg/kg ( P ＜0.05 ). The volume of spinal cord astrocytes cultured in vitro after OGD 6 h/R 24 h was significantly greater than that after OGD 6 h/R 6 h or OGD 6 h/R 12 h ( P ＜0.05 ). EP at 12 μmol/L reduced cell swelling, decreased expression of HMGB1, AQP4 and TLR4, and downgraded nuclear expression of NF-κB in spinal cord astrocytes after OGD/R significantly more than EP at 6 μmol/L( P ＜0.05). Conclusion EP may reduce early spinal cord edema after SCI, attenuate spinal cord astrocyte swelling and decrease AQP4 expression after OGD/R in vitro by inhibiting HMGB1 in rats.
ABSTRACT
Objective·To observe the degree of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) at different time points, and to investigate the effects of ethyl pyruvate (EP) on CVS following SAH and its mechanism. Methods·Fifty healthy SD rats were randomly divided into five groups i.e. sham group, SAH-3 d group, SAH-5 d group, SAH-7 d group and SAH-9 d group (n=10 in each group). Sham group was built by double cisternal injection with 0.3 mL saline each time, and SAH group was built by double cisternal injection with 0.3 mL autologous blood each time. The neurological function and degree of CVS were observed. Another forty-eight healthy SD rats were randomly divided into three groups i.e. sham+saline (equal volume) group, SAH+saline (equal volume) group and SAH+EP [100 mg/(kg·d)] group (n=16 in each group). The neurological function, degree of CVS and cell apoptosis of basilar artery were observed on day 7. The expressions of p-Akt, Bax, Bcl-2 and cleaved caspase-3 were also observed on day 7. Results·CVS significantly increased on day 3, decreased on day 5, and then significantly increased to top level on day 7, and gradually decreased on day 9. Compared with sham+saline group, the neurological scores were significantly decreased in SAH+saline group on day 7. Compared with sham+saline group, CVS and cell apoptosis of basilar artery were significantly increased in SAH+saline group on day 7. Compared with sham+saline group, the expressions of Bax and cleaved caspase-3 were significantly up-regulated, while the expressions of p-Akt and Bcl-2 were significantly down-regulated in SAH+saline group on day 7. Compared with SAH+saline group, the neurological scores significantly increased in SAH+EP group on day 7. Compared with SAH+saline group, CVS and cell apoptosis of basilar artery significantly decreased in SAH+EP group on day 7. Compared with SAH+saline group, the expressions of Bax and cleaved caspase-3 were significantly down-regulated, while the expressions of p-Akt and Bcl-2 were significantly up-regulated in SAH+EP group on day 7. Conclusion·The double hemorrhage model rat has most severely CVS on day 7. EP can attenuate vasospasm after SAH, and its mechanism may be associated with inhibition of cell apoptosis. PI3K/Akt signaling pathway may be involved, which may provide a novel therapeutic target for CVS.
ABSTRACT
Objective To investigate the impact of ethyl pyruvate (EP) on cognitive function and the expression of high mobility group box 1 (HMGB1) and receptor for advanced glycation end products (RAGE) after splenectomy in aged rats.Methods Eighty-four male aged Sprague-Dawley rats, 18 months old, weighing 500-600 g, were randomly divided into 4 groups (n=21 each) by random number table method: control group (group C), surgery group (group S) ethyl pyruvate group (group E) and solution without EP group (group R).Morris water maze test was performed to evaluate cognitive function 5 days before surgery and 1, 3, 7 days after surgery.Group E was injected with EP 40 mg/kg intrapertoneally after splenectomy, group S and group C were injected with equivalent normal saline after splenectomy, group R was injected with equivalent solution without EP.Rats were killed after Morris water maze test, and the expression of HMGB1 and RAGE protein and mRNA in hippocampus were measured by Western blot and RT-PCR methods.Results Compared with group C, the escape latency and swimming distance were significantly prolonged in groups S, E and R 1 and 3 days after surgery, as well as the expression of HMGB1 and RAGE in hippocampus were significantly up-regulated (P＜0.05).Compared with group S, the escape latency and swimming distance were significantly decreased and the expression of HMGB1 and RAGE were down-regulated in group E 1 and 3 days after surgery (P＜0.05).Compared with the preoperative group, the escape latency and swimming distance were significantly prolonged in groups S, E and R 1 and 3 days after surgery (P＜0.05).Conclusion EP may improve cognitive function in aged rats by down regulating the expression of HMGB1 and RAGE in the hippocampus.
ABSTRACT
<p><b>INTRODUCTION</b>The integration of reactive oxygen species is strongly associated with important pathophysiological mechanisms that mediate myocardial ischaemia/reperfusion (I/R) damage. Pyruvate is an efficacious scavenger of reactive oxygen species and a previous study has shown that ethyl pyruvate (EP) has a myocardial protective effect against regional I/R damage in an in vivo rat model. The purpose of this study was to determine whether the myocardial protective effect of EP is associated with anti-apoptosis.</p><p><b>METHODS</b>Rats were allocated to receive EP dissolved in lactated Ringer's solution or lactated Ringer's solution alone, via intraperitoneal infusion one hour before ischaemia. They were exposed to 30 minutes of ischaemia followed by reperfusion of the left coronary artery territory over two hours. Anti-apoptotic effects were checked using several biochemical parameters after two hours of reperfusion. Apoptosis was analysed using measured caspase-3 activity, Western blotting of B-cell lymphoma 2 (Bcl-2) family protein cleaved by caspase-3, and assessment of DNA laddering patterns and the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining test.</p><p><b>RESULTS</b>In ischaemic myocardium, EP increased Bcl-2 expression, but reduced Bcl-2-associated X protein and cleaved caspase-3 expressions. EP reduced the expression of DNA laddering and the number of myocardial I/R-damaged TUNEL-positive cells.</p><p><b>CONCLUSION</b>This study demonstrated that EP has an anti-apoptotic effect after regional I/R damage in an in vivo rat heart model. The myocardial protective effect of EP may be related to its anti-apoptotic effect.</p>
ABSTRACT
Entre los radioprotectores con uso clínico se destaca la amifostina (WR- 2721), eficaz pero con efectos secundarios que impiden su uso repetitivo. Es interés de los autores desarrollar radioprotectores menos tóxicos, por sí mismos o como coadyuvantes de amifostina. Ratas machos o hembras se expusieron a una dosis de rayos X de 2 Gy. Se ensayó el piruvato de etilo, solo o conjuntamente con amifostina. Cuarenta y ocho horas después de la exposición a la radiación, se realizó el recuento de eritrocitos, de leucocitos y la fórmula leucocitaria. Los efectos genotóxicos se evaluaron en leucocitos de sangre mediante el ensayo Cometa. Se realizaron también estudios de supervivencia a 60 días post-irradiación. En los animales irradiados disminuyeron los eritrocitos, y el recuento de leucocitos se redujo drásticamente respecto al control, presentando además una fórmula alterada. El tratamiento con piruvato de etilo resultó en una protección de los eritrocitos en ambos sexos. El daño genético disminuyó significativamente por el tratamiento con piruvato de etilo solo o combinado con amifostina, y en hembras se observó una mayor supervivencia solo con el tratamiento combinado. El piruvato de etilo mostró una acción radioprotectora significativa, que podría mejorarse aumentando la dosis o el tiempo de tratamiento, ya que tiene muy baja toxicidad.
Among the currently available radioprotectors, only amifostine (WR-2721) has shown in clinical trials to reduce radiation-induced toxicity. This compound is an efficient radioprotector but it exhibits some undesirable side effects which prevent its repetitive use. Efforts are directed to develop radioprotective agents with lower toxicity, with their own protective potential or suitable as coadyuvants of amifostine. The present study describes the results obtained by repetitive oral administration of ethyl pyruvate. Male or female rats were exposed to an X-ray dose of 2 Gy. Forty-eight hours after exposure to radiation, erythrocyte count, leukocyte and differential count were performed. Genotoxic effects were assessed in blood leukocytes by the Comet assay. Survival studies were also performed at 60 days post-irradiation. Eritrocyte and leukocyte were reduced in animals exposed to radiation compared to the control, also presenting an altered formula. Treatment with ethyl pyruvate resulted in a protection on erythrocytes of both sexes. Genetic damage was significantly decreased by ethyl pyruvate alone or combined with amifostine, and in females, higher survival was observed only with combined administration. Ethyl pyruvate showed a significant radioprotective action, which could be improved by increasing the dose or time of treatment because it has low toxicity.
Entre os radioprotetores com uso clínico destaca-se a amifostina (WR-2721) eficaz mas com efeitos secundários que impedem seu uso repetitivo. O interesse dos autores é desenvolver radioprotetores menos tóxicos, por si mesmos ou como coadjuvantes de amistofina. Ratos machos ou fêmeas foram expostos a doses de raios X de 2Gy. Ensaiou-se o piruvato de etila, só ou junto com amifostina. Quarenta e oito horas após a exposição à radiação foi realizada a contagem de eritrócitos, de leucócitos e da fórmula leucocitária. Efeitos genotóxicos foram avaliados em leucócitos do sangue pelo Ensaio Cometa. Estudos de sobrevivência foram também realizados a 60 dias pós-irradiação. Nos animais irradiados diminuíram os eritrócitos, e a contagem de leucócitos se reduziu drasticamente em comparação com o controle, apresentando também uma fórmula alterada. O tratamento com piruvato de etila resultou numa proteção dos eritrócitos em ambos os sexos. O dano genético diminuiu significativamente pelo tratamento com piruvato de etila sozinho ou combinado com amifostina, e nas fêmeas se observou maior sobrevivência só com o tratamento combinado. O piruvato de etila mostrou uma ação radioprotetora significativa, que poderia ser melhorada pelo aumento da dose ou do tempo de tratamento, visto que tem baixa toxicidade.
Subject(s)
Rats , Amifostine/toxicity , Radiation , Radiation-Protective Agents/therapeutic use , Amifostine/administration & dosage , Therapeutics/statistics & numerical dataABSTRACT
Objective:To establish a method for the determination of ethyl pyruvate by GC. Methods:A Varian CP7502 capillary column(25 m × 0. 25 mm,0. 25 μm)was used. The carrier gas was nitrogen with the flow rate of 30 ml·min-1 ,the gas was hydrogen with the flow rate of 40 ml·min -1 and the oxidant gas was air with the flow rate of 400 ml·min-1 . The detector was an FID and the inlet temperature was 210℃ . The temperature program was as follows:the initial column temperature was 40℃, and then risen to 200℃ with a rate of 15 ℃·min -1 . The split ratio was 100 ∶1 and the injection volume was 1μl. Results:Ethyl pyruvate had a good linear relationship within the range of 0. 025 6- 8. 192 8 mg·ml -1 ,and r was 0. 999 8. The average recovery was99. 99% and RSD was 1. 38%(n = 9). Conclusion:The method is accurate,simple and rapid with good stability and high sensitivity,which can be used to determine the content of ethyl pyruvate.
ABSTRACT
Objective To investigate the effect of ethyl pyruvate (EP) on the expression of caspase-3 in the hippocampus and the learn-ing and memory ability in neonatal rats with hypoxic ischemic brain damage (HIBD). Methods 90 Wistar rats of 7 days old were randomly divided into sham operation group (n=15), HIBD group (n=15), EP1 group (n=15, 10 ml/kg), EP2 group (n=15, 30 mg/kg), EP3 group (n=15, 50 mg/kg) and EP4 group (n=15, 100 mg/kg). The model was established with Rice's method. 30 minutes before operation, and every 24 hours after operation, EP groups were injected with 10 ml/kg, 30 mg/kg, 50 mg/kg, 100 mg/kg EP in abdomen respectively, for 2 weeks. Af-ter treatment, the caspase-3 positive cells were observed by immunohistochemical staining, and the latency and the times crossing the target quadrant were tested by Morris water maze test. Results The caspase-3 positive cells were less in EP groups than in HIBD group (P0.05), especially in EP3 group (P0.05), especially in EP3 group (P<0.01). Conclusion Ethyl pyruvate can de-crease the expression of caspase-3 in hippocampus, and improve the ability of memory and learning in neonatal rats with HIBD.
ABSTRACT
OBJECTIVES: Experimental studies on sepsis have demonstrated that ethyl pyruvate is endowed with antioxidant and anti-inflammatory properties. This study aimed to investigate the effects of ethyl pyruvate on leukocyte-endothelial interactions in the mesenteric microcirculation in a live Escherichia coli-induced sepsis model in rats. METHODS: Male Wistar rats were administered an intravenous suspension of E. coli bacteria or were subjected to a sham procedure. Three hours after bacterial infusion, the rats were randomized into the following groups: a control group without treatment, a group treated with lactated Ringer’s solution (4 mL/kg, i.v.), and a group treated with lactated Ringer’s solution (4 mL/kg, i.v.) plus ethyl pyruvate (50 mg/kg). At 24 h after bacterial infusion, leukocyte-endothelial interactions were investigated using intravital microscopy, and the expression of P-selectin and intercellular adhesion molecule-1 was evaluated via immunohistochemistry. White blood cell and platelet counts were also determined at baseline and 3 h and 24 h after E. coli inoculation. RESULTS: The non-treated and lactated Ringer’s solution-treated groups exhibited increases in the numbers of rolling leukocytes (∼2.5-fold increase), adherent cells (∼3.0-fold), and migrated cells (∼3.5-fold) compared with the sham group. In contrast, treatment with Ringer’s ethyl pyruvate solution reduced the numbers of rolling, adherent and migrated leukocytes to the levels observed in the sham group. Additionally, the expression of P-selectin and intercellular adhesion molecule-1 was significantly increased on mesenteric microvessels in the non-treated group compared with the sham group (p<0.001). The expression of both adhesion molecules was reduced in the other groups, with ethyl pyruvate being more effective than lactated Ringer’s solution. Infusion of bacteria caused significant leukopenia (3 h), followed ...
Subject(s)
Animals , Male , Rats , Cell Communication/drug effects , Endothelial Cells/drug effects , Leukocytes/drug effects , Mesenteric Veins/drug effects , Pyruvates/pharmacology , Sepsis/drug therapy , Cell Communication/physiology , Disease Models, Animal , Escherichia coli Infections , Endothelial Cells/cytology , Leukocytes/cytology , Microcirculation , Mesenteric Veins/cytology , Rats, WistarABSTRACT
INTRODUÇÃO: Estudos recentes em modelos experimentais de sepse demonstraram as propriedades antioxidante e anti-inflamatória do etilpiruvato. Diferentes modelos experimentais também demonstraram que pequenos volumes de solução salina hipertônica (7,5%) melhoram a hemodinâmica, a microcirculação e modulam o sistema imunológico. Este estudo teve como objetivo investigar os efeitos do etil-piruvato, da solução salina hipertônica e da solução de Ringer lactato sobre a microcirculação mesentérica em modelo de sepse induzida por Escherichia coli viva em ratos. MÉTODOS: Ratos Wistar machos receberam por via endovenosa uma suspensão de E. coli ou foram submetidos ao procedimento cirúrgico do grupo falso-operado. Após três horas da infusão bacteriana os animais foram randomizados em: grupo controle não tratado, grupo tratado com solução de Ringer lactato (4mL/kg i.v.); grupo tratado com solução de Ringer lactato (4 mL/kg i.v.) associado a etil-piruvato (50mg/kg) e grupo tratado com solução salina hipertônica (7,5%, 4 mL/kg i.v.). Após 24 horas da bacteremia, as interações leucócito-endotélio foram investigadas por microscopia intravital, e a expressão de P-selectina e da molécula de adesão intercelular (ICAM)-1 determinada por imuno-histoquímica. Leucograma e contagem de plaquetas foram realizadas no início do estudo, 3 horas e 24 horas após a inoculação de E. coli. RESULTADOS: Os grupos não tratado e tratado com solução de Ringer lactato exibiram um aumento no número de leucócitos rollers (~ 2,5 vezes), leucócitos aderidos (~ 3,0 vezes), e de leucócitos migrados (~ 3,5 vezes) comparados ao grupo falso operado. O tratamento com etil-piruvato reduziu o número de leucócitos rollers, aderidos e migrados aos níveis obtidos no grupo falso operado (p > 0,05). Efeitos semelhantes foram observados nos animais tratados com a solução salina hipertônica (p > 0,05). A expressão de P-selectina e de ICAM-1 aumentou significativamente na microcirculação mesentérica no grupo...
BACKGROUND: Experimental studies on sepsis have demonstrated that ethyl pyruvate is endowed with antioxidant and anti-inflammatory properties. It has been shown that small volumes of hypertonic saline solution (7.5%) improve hemodynamics, the microcirculation, and modulate the immune system. This study aimed to investigate the effects of ethyl pyruvate, hypertonic saline and lactated Ringer's solution on mesenteric microcirculation in a sepsis model induced by live Escherichia coli in rats. METHODS: Male Wistar rats were underwent an intravenous suspension of E. coli bacteria or submitted to the sham procedure. After 3h of bacteria infusion rats were randomized into: control, without treatment; treated with lactated Ringer's solution (4 mL/kg, i.v.); treated with lactated Ringer's solution (4mL/kg, i.v.) plus ethyl pyruvate (50mg/kg), and treated with hypertonic saline solution (7.5%, 4 mL/kg i.v.). At 24h after bacteria infusion leukocyte-endothelial interactions were investigated by intravital microscopy, and the expression of P-selectin and intercellular adhesion molecule (ICAM)- 1 evaluated by immunohistochemistry. White blood cell and platelet counts were determined at baseline, 3h and 24h after E. coli inoculation. RESULTS: Both non-treated and lactated Ringer's-treated groups exhibited an increase in the number of rolling leukocytes (~2.5-fold), adherent (~3.0-fold), and migrated cells (~3.5-fold) compared to sham. Treatment with Ringer's ethyl pyruvate solution reduced the number of rolling, adherent and migrated leukocytes to the levels attained in the sham group (p > 0.05). Similar effects were observed when animals were treated with hypertonic saline (p > 0.05). The expression of P-selectin and ICAM-1 significantly increased on mesenteric microvessels in non-treated group compared with sham (p < 0.001). All treatments reduced the expression of both adhesion molecules being ethyl pyruvate and hypertonic saline solution...
Subject(s)
Animals , Male , Rats , Escherichia coli , Intercellular Adhesion Molecule-1 , Microcirculation , P-Selectin , Pyruvates , Rats, Wistar , Saline Solution, Hypertonic , SepsisABSTRACT
Ethyl pyruvate ( EP) is a stable lipophilic ester derivative of pyruvate with many pharmacological actions confirmed by researches , the anti-tumor effects of EP has attracted the attention of many domestic and foreign scholars .EP exerts antitumor activi-ty through several ways , such as inhibiting proliferation , inducing apoptosis , inhibiting angiogenesis , blocking cell cycle .This paper briefly reviews the rencent findings of antitumor function of EP .
ABSTRACT
Objective To explore the role of ethyl pyruvate (EP) on E-cadherin of airway epithelium and airway inflammation in a TDI-induced mouse asthma model. Methods 30 male BALB/c mice were randomly divided into control group , asthma group and EP group. On day 1 and 8 , mice in asthma group and EP group were treated with 0.3%TDI on the dorsum of both ears for sensitization. And on day 15 , 18 and 21 the mice underwent an aerosol inhalation of 3% TDI, and saline (100 mg/kg) was injected intraperitoneally 1 hour before inhalation. The control group underwent acetone and olive oil (AOO) sensitization on day 1 and 8, AOO challenge on day 15, 18 and 21. Saline (100 mg/kg) was injected intraperitoneally 1 hour before challenge. One hour before each challenge, mice were given EP (100mg/kg) or vehicle via intraperitoneal injection. On day 22, airway reactivity, IL-4 , IFN-γand IgE in the serum were detected , immunohistochemistry and WB were used to assess E-cadherin levels. Results Airway reactivity, IL-4, IFN-γin and IgE in the serum in asthma group are significantly higher than that in control group (P<0.05). Treatment with EP dramatically decreased airway hyperresponsiveness in TDI-challenged mice, as well as IL-4, IFN-γ and IgE (P < 0.05). E-cadherin in control group was distributed evenly at the connection of epithelial cells. E-cadherinin distribution was chaotic and its expression was decreased in asthma group. EP intervention can ameliorate the damage of E-cadherinin. Conclusions EP can ameliorate the destruction of E-cadherin in airway epithilum by TDI.
ABSTRACT
BACKGROUND: Although paclitaxel is a widely used chemotherapeutic agent for the treatment of solid cancers, side effects such as neuropathic pain lead to poor compliance and discontinuation of the therapy. Ethyl pyruvate (EP) is known to have analgesic effects in several pain models and may inhibit apoptosis. The present study was designed to investigate the analgesic effects of EP on mechanical allodynia and apoptosis in dorsal root ganglion (DRG) cells after paclitaxel administration. METHODS: Rats were randomly divided into 3 groups: 1) a control group, which received only vehicle; 2) a paclitaxel group, which received paclitaxel; and 3) an EP group, which received EP after paclitaxel administration. Mechanical allodynia was tested before and at 7 and 14 days after final paclitaxel administration. Fourteen days after paclitaxel treatment, DRG apoptosis was determined by activated caspase-3 immunoreactivity (IR). RESULTS: Post-treatment with EP did not significantly affect paclitaxel-induced allodynia, although it tended to slightly reduce sensitivities to mechanical stimuli after paclitaxel administration. After paclitaxel administration, an increase in caspase-3 IR in DRG cells was observed, which was co-localized with NF200-positive myelinated neurons. Post-treatment with EP decreased the paclitaxel-induced caspase-3 IR. Paclitaxel administration or post-treatment with EP did not alter the glial fibrillary acidic protein IRs in DRG cells. CONCLUSIONS: Inhibition of apoptosis in DRG neurons by EP may not be critical in paclitaxel-induced mechanical allodynia.
Subject(s)
Animals , Rats , Apoptosis , Caspase 3 , Compliance , Diagnosis-Related Groups , Ganglia, Spinal , Glial Fibrillary Acidic Protein , Hyperalgesia , Myelin Sheath , Neuralgia , Neurons , Paclitaxel , Pyruvates , Pyruvic AcidABSTRACT
BACKGROUND: It has been demonstrated that the expression of tumor necrosis factor-alpha (TNF-alpha) and apoptotic cell death in the dorsal root ganglion (DRG) following spinal nerve constriction injury play a role in the initiation and continuation of hyperalgesia and allodynia. The present study was designed to investigate the effects of ethyl pyruvate (EP) on mechanical and cold allodynia, TNF-alpha expression, and apoptosis in DRG after spinal nerve ligation injury. METHODS: Rats were divided into 3 groups: control, pre-EP, and post-EP. EP (50 mg/kg) was intraperitoneally injected 30 minutes before (pre-EP) or after (post-EP) surgery. Behavioral tests to determine mechanical and cold allodynia were conducted before surgery and 4 and 7 days after surgery. Seven days after surgery, TNF-alpha protein levels in DRG were evaluated by enzyme-linked immunosorbent assay, and DRG apoptosis was determined by immunohistochemical detection of activated caspase-3. RESULTS: Treatment with EP significantly reduced mechanical and cold allodynia following spinal nerve ligation injury. TNF-alpha protein levels in the pre-EP (4.7 +/- 1.2 pg/200 microg; P < 0.001) and post-EP (6.4 +/- 1.8 pg/200 microg; P < 0.001) groups were 2-3 times lower than the control group (14.4 +/- 1.2 pg/200 microg). The percentages of neurons and satellite cells that co-localized with caspase-3 were also significantly lower in the pre-EP and post-EP groups than the control group. CONCLUSIONS: These results demonstrate that EP has a strong anti-allodynic effect that acts through the inhibition of TNF-alpha expression and apoptosis in DRG after spinal nerve ligation injury.
Subject(s)
Animals , Rats , Apoptosis , Caspase 3 , Cell Death , Cold Temperature , Constriction , Diagnosis-Related Groups , Enzyme-Linked Immunosorbent Assay , Ganglia, Spinal , Hyperalgesia , Ligation , Neurons , Pyruvates , Pyruvic Acid , Spinal Nerve Roots , Spinal Nerves , Tumor Necrosis Factor-alphaABSTRACT
Hemorrhagic hypovolemic shock secondary to trauma is an important cause of morbidity and mortality worldwide. During the last few years, new concepts have emerged and the guidelines of fluid resuscitation in these patients have been redefined. The concept of hypotensive resuscitation has been established and new colloid solutions based on starch have been manufactured, been hydroxyethyl starch in a balanced electrolytic solution, the most studied and successful one. It has been reported, as well, the positive effects of the pharmacologic modulation of the inflammatory pathways in experimental model subjects submitted to hypovolemic shock. Products such as, ethyl pyruvate and the Na+/H+ type 1 inhibitor, BIIB513, have been Studies only experimentally in rodent models using colloids as the primary resuscitation fluid. The significant improvement in the hemodinamyc, pattern and the cardiac and inflammatory indexes and mediators, has created the basis for their use in clinical trials in the near future. The systemic inflammatory response is an important cause of multiple organ failure that increases the late mortality of patients surviving the initial early phases of hypovolemic traumatic shock and its experimental modulation in rodent models with products such as ethyl pyruvate and BIIB513 has produced excellent in vivo and in vitro results.
Universalmente se considera el Shock hipovolémico de origen hemorrágico como una importante causa de morbi-mortalidad. Durante los últimos años se ha redefinido los conceptos de la reanimación con líquidos intravenosos en los pacientes con choque hipovolémico y establecido los conceptos de reanimación hipotensa con el uso de nuevos coloides derivados del almidón, tales como el hidroxietil-almidón en solución electrolítica balanceada (Hextend®). Así mismo, se ha reportado el beneficio que conlleva el uso de modificadores de la cascada inflamatoria en modelos experimentales de sujetos sometidos a choque hipovolémico hemorrágico. Productos como el etil piruvato y la BIIB513, un inhibidor selectivo del intercambiador Na+/H+ tipo 1, han sido estudiados sólo experimentalmente en modelos roedores, empleando coloides como principal elemento de reanimación. Al mejorar el perfil hemodinámico, parámetros cardíacos y niveles de mediadores inflamatorios, estos compuestos constituyen una base cierta para ser incluidos en estudios clínicos en un futuro próximo. La respuesta inflamatoria sistémica está íntimamente implicada en la patogénesis de la Falla Orgánica Múltiple, aumentando la mortalidad tardía de pacientes que sobreviven las etapas tempranas del shock hipovolémico hemorrágico traumático. Su modulación experimental con el etil piruvato o bien la BIIB513 ha dado excelente resultado tanto en modelos experimentales in vivo como in vitro.
Subject(s)
Humans , Sodium-Hydrogen Exchangers/antagonists & inhibitors , Hydroxyethyl Starch Derivatives/pharmacology , Mesylates/pharmacology , Shock/drug therapy , Isotonic Solutions/pharmacology , Hemodynamics , Wounds and Injuries/complications , Inflammation , Resuscitation/methods , Shock, Hemorrhagic/etiology , Shock, Hemorrhagic/drug therapy , Shock/etiology , Plasma Substitutes/pharmacologyABSTRACT
PURPOSE: Ethyl pyruvate has anti-inflammatory properties and protects organs from ischemia/reperfusion (I/R)-induced tissue injury. The aim of this study was to determine whether ethyl pyruvate decreases the inflammatory response after regional I/R injury and whether ethyl pyruvate protects against delayed regional I/R injury in an in vivo rat heart model after a 24 hours reperfusion. MATERIALS AND METHODS: Rats were randomized to receive lactated Ringer's solution or ethyl pyruvate dissolved in Ringer's solution, which was given by intraperitoneal injection 1 hour prior to ischemia. Rats were subjected to 30 min of ischemia followed by reperfusion of the left coronary artery territory. After a 2 hours reperfusion, nuclear factor kappaB, myocardial myeloperoxidase activity, and inflammatory cytokine levels were determined. After the 24 hours reperfusion, the hemodynamic function and myocardial infarct size were evaluated. RESULTS: At 2 hours after I/R injury, ethyl pyruvate attenuated I/R-induced nuclear factor kappaB translocation and reduced myeloperoxidase activity in myocardium. The plasma circulating levels of inflammatory cytokines decreased significantly in the ethyl pyruvate-treated group. At 24 hours after I/R injury, ethyl pyruvate significantly improved cardiac function and reduced infarct size after regional I/R injury. CONCLUSION: Ethyl pyruvate has the ability to inhibit neutrophil activation, inflammatory cytokine release, and nuclear factor kappaB translocation. Ethyl pyruvate is associated with a delayed myocardial protective effect after regional I/R injury in an in vivo rat heart model.
Subject(s)
Animals , Male , Rats , Anti-Inflammatory Agents/pharmacology , Cell Nucleus/metabolism , Cytoplasm/metabolism , Heart/physiopathology , Inflammation , Myocardial Infarction/prevention & control , Myocardium/metabolism , NF-kappa B/metabolism , Peroxidase/metabolism , Pyruvates/pharmacology , Rats, Sprague-Dawley , Reperfusion Injury/drug therapyABSTRACT
In order to study the effects of ethyl pyruvate on cardiomyocyte apoptosis following ischemia/reperfusion (I/R) in vitro and the expression of Bcl-2 and Bax proteins, isolated rat hearts were perfused in a Langendorff model. Twenty-four rats were randomly divided into 3 groups (n=8 in each group): control group was perfused for 120min. In the I/R group, after 30min stabilization the injury was induced by 30min global ischemia followed by 60min reperfusion. Ethyl pyruvate (EP) group was set up with the same protocol as I/R group except that it was supplied with 2mmol/L EP 15min before ischemia and throughout reperfusion. Myocardial malonaldehyde (MDA) content Was measured. Myocardial apoptotic index (AI) was tested by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. The expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax in cardiac myocytes was detected by immunohistochemistry. As compared with control group, the content of MDA, myocardial AI and the expression of Bcl-2, Bax proteins were increased significantly in I/R group, but the content of MDA, myocardial AI and the expression of Bax protein were decreased obviously and the expression of Bcl-2 protein was up-regulated in EP group (P<0.05). These results demonstrate that EP could inhibit apoptosis of cardiac myocytes possibly via alleviating oxidative stress, up-regulating Bcl-2 and down-regulating Bax proteins.
ABSTRACT
Objective To explore the effects of ethyl pyruvate (EP) on hepatic high mobility group box-1 protein (HMGB1) expression in experimental routine with acute necrotizing pancreatitis (ANT). Method ANP model was induced by retrograde injection of 5 % sodium taurocholate into pancreatic duct. Twenty-four male wistar rats were divided randomly into 3 groups(8 rats in each group): group A (ANT group); group B (ANP rats re-ceived ethyl pyruvate therapy) and group C (control group with sham operation). The concentration of plasma amylase (AMY), A.sr and ALT, and the activity of myeloperoxidase (MPO) in the liver were determined. The ex-pression of HMGB1 mRNA in liver was detected by using reverse transcription polymerase chain reaction (RT-PCR). The changes of morphological damage were observed under microscopy. The expression of HMGB1 in the liver was observed by using SP immunohistochemistry. ANOVA was performed with SPSS 10.0 statistical analysis software and the difference was accepted as significant if the P<0.05, as verified by using Duncan's and Tukey' s post hoc test. Results Compared with gxoup A,levels of plasma AMY,AST and ALT in group B were markedly lower (P<0.05). Compared with group C, MPO in group A was higher significantly (P<0.01).with group A, the pathological changes of pancreas and liver in group B were milder. Compared with group C,the hepatic HMGB1 mRNA expression was markedly higher in group A [(0.28±0.04) vs. (0.73±0.06), P<0.01]. By contrast,the HMGB1 mRNA expression was markedly lower in group B compared with group A [(0.46±0.05) vs. (0.73±0.06), P<0.05]. The HMGB1 protein expression in hepatocytes and Kupffer's cells of rats with ANP was significantly up-regulated compared with control group, but it was reduced significantly in EP treatment group. Conclusions HMGB1 as a late mediator in liver might be involved in the pathogenesis of acute hepatic injury with ANP. EP could down-regulate the hepatic HMGB1 expression together with improvement of liver function in rats with ANP.