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1.
Article in Chinese | WPRIM | ID: wpr-880089

ABSTRACT

OBJECTIVE@#To investigate the influence of GPT2(glutamic pyruvate transaminase 2)to biological characteristics of human acute myeloid leukemia cell line HL-60.@*METHODS@#The expression of GPT2 in hematological tumor and AML cell was detected. The lentvirus-mediated of short-hairpin RNA (shRNA) was constricted, and the knock-down efficiency of HL-60 in AML cell after infected by lentvirus-mediated was detected by Western blot and Q-PCR. CCK-8 assay and soft agar colony formation assay were used to detect the effect of GPT2 gene deletion to the cell proliferation potential. Fluorescence activated cell sorting(FACS) was used to analyze the effect of gene deletion to the cell cycle and Caspase 3/7 Activity Assay Kit was used to analyze the effect of GPT2 gene deletion to the cell apoptosis.@*RESULTS@#GPT2 showed mRNA high expression in AML patients. CCK-8, soft agar assay, and Caspase 3/7 Activity Assay Kit results showed that compared with shCtrl group, the cells in shGPT2-1、shGPT2-2、shGPT2-3 group showed the slowing down on proliferation, decreasing on colony ability, and the apoptosis of the cells was increasing significantly. FACS showed that GPT2 gene was related to the cycle of HL-60 cell.@*CONCLUSION@#GPT2 appears to involve the proliferation, cycle distribution and apoptosis of AML cell HL-60. The deletion of GPT gene can lead to the inhibitation of cells proliferation and increase apoptosis.


Subject(s)
Apoptosis , Cell Proliferation , HL-60 Cells , Humans , Leukemia, Myeloid, Acute , Pyruvates , Transaminases
2.
Acta cir. bras ; 31(11): 724-729, Nov. 2016. graf
Article in English | LILACS | ID: biblio-827660

ABSTRACT

ABSTRACT PURPOSE: To investigate the hepatotoxicity and nephrotoxicity of 3-Bromopyruvate (3BP) in mice. METHODS: Fifteen nude mice were grafted subcutaneously in the left flank with MDA-MB-231 cells, then all mice were divided into control group (PBS), 3BP group (8 mg/kg), positive group (DNR: 0.8 mg/kg) when tumor volume reached approximately 100 mm3. 28 days later, tumors, livers and kidneys were stored in 4 % formalin solution and stained with hematoxylin and eosin staining. The Kunming mice experiment included control group (PBS), 3BP group (4mg/kg; 8mg/kg; 16mg/kg), positive group (DNR: 0.8 mg/kg). 24 hours later, the blood were used for the determination of hepatic damage serum biomarkers. Livers were stored in 4 % formalin solution for the later detection. RESULTS: 3BP at the dose of 8mg/kg had a good effect on inhibiting tumor growth in nude mice and did not damage liver and kidney tissues. Kunming mice experiment showed 3BP at the dose of 16mg/kg did damage to liver tissues. CONCLUSION: 3-Bromopyruvate at the dose of suppressing tumor growth did not exhibit hepatotoxicity and nephrotoxicity in nude mice, and the effect on liver was confirmed in Kunming mice.


Subject(s)
Animals , Female , Mice , Pyruvates/toxicity , Enzyme Inhibitors/toxicity , Chemical and Drug Induced Liver Injury/pathology , Acute Kidney Injury/pathology , Kidney/drug effects , Liver/drug effects , Disease Models, Animal , Dose-Response Relationship, Drug , Acute Kidney Injury/chemically induced , Liver Neoplasms, Experimental/drug therapy , Mice, Inbred BALB C , Mice, Nude
3.
Clinics ; 70(7): 508-514, 2015. tab, graf
Article in English | LILACS | ID: lil-752397

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, Wistar
4.
São Paulo; s.n; 2014. [84] p. ilus, tab, graf.
Thesis in Portuguese | LILACS | ID: lil-748486

ABSTRACT

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 , Sepsis
5.
Chinese Journal of Hematology ; (12): 637-640, 2014.
Article in Chinese | WPRIM | ID: wpr-242098

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of glycolytic inhibitor 3-Bromopyruvate (3-BrPA) on the proliferation and apoptosis of mouse spleen lymphocytes and explore its mechanism.</p><p><b>METHODS</b>An one-way mixed lymphocyte culture (MLC) system was established, including BALB/c mouse spleen cells (H-2d) as stimulator and C57BL/6 mouse spleen cells (H-2b) as responder. With treatment of 3-BrPA at different concentrations (0-200 μmol/L), lymphocyte proliferation capacity was detected by the CCK-8 method, the expression of CD3, CD4, and CD8 by flow cytometry, and the concentrations of cytokine interleukin (IL)-4 and interferon (IFN)-γ in the supernatant by ELISA.</p><p><b>RESULTS</b>At a middle or high dose (over 20 μmol/L), 3-BrPA displayed a dose-dependent inhibitory effect on lymphocyte proliferation in the MLC system. The 50% inhibitory concentration (IC50) were 48.6, 41.2, and 41.9 μmol/L after 24, 36, and 48 h culture, respectively. With treatment of 50 μmol/L 3-BrPA, the IFN-γ level [(164.25 ± 20.14) ng/L] was significantly lower, compared with control [(277.61 ± 18.46) ng/L]. The IL-4 level [(31.06 ± 6.06) ng/L] was significantly higher, compared with control [(28.64 ± 3.97) ng/L]. Consequently, the IFN-γ/IL-4 ratio decreased significantly.</p><p><b>CONCLUSION</b>These results indicate that 3-BrPA had a significant inhibitory effect on the proliferation of mouse spleen lymphocytes cultured in MLC system, accompanied with the Th2-biased secretion of cytokines.</p>


Subject(s)
Animals , Apoptosis , Cell Proliferation , Cells, Cultured , Interferon-gamma , Metabolism , Interleukin-4 , Metabolism , Lymphocyte Culture Test, Mixed , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Pyruvates , Pharmacology , Spleen , Cell Biology , Metabolism
6.
Chinese Journal of Cancer ; (12): 356-364, 2014.
Article in English | WPRIM | ID: wpr-320524

ABSTRACT

3-Bromopyruvate (3BP) is a new, promising anticancer alkylating agent with several notable functions. In addition to inhibiting key glycolysis enzymes including hexokinase II and lactate dehydrogenase (LDH), 3BP also selectively inhibits mitochondrial oxidative phosphorylation, angiogenesis, and energy production in cancer cells. Moreover, 3BP induces hydrogen peroxide generation in cancer cells (oxidative stress effect) and competes with the LDH substrates pyruvate and lactate. There is only one published human clinical study showing that 3BP was effective in treating fibrolamellar hepatocellular carcinoma. LDH is a good measure for tumor evaluation and predicts the outcome of treatment better than the presence of a residual tumor mass. According to the Warburg effect, LDH is responsible for lactate synthesis, which facilitates cancer cell survival, progression, aggressiveness, metastasis, and angiogenesis. Lactate produced through LDH activity fuels aerobic cell populations inside tumors via metabolic symbiosis. In melanoma, the most deadly skin cancer, 3BP induced necrotic cell death in sensitive cells, whereas high glutathione (GSH) content made other melanoma cells resistant to 3BP. Concurrent use of a GSH depletor with 3BP killed resistant melanoma cells. Survival of melanoma patients was inversely associated with high serum LDH levels, which was reported to be highly predictive of melanoma treatment in randomized clinical trials. Here, we report a 28-year-old man presented with stage IV metastatic melanoma affecting the back, left pleura, and lung. The disease caused total destruction of the left lung and a high serum LDH level (4,283 U/L). After ethics committee approval and written patient consent, the patient received 3BP intravenous infusions (1-2.2 mg/kg), but the anticancer effect was minimal as indicated by a high serum LDH level. This may have been due to high tumor GSH content. On combining oral paracetamol, which depletes tumor GSH, with 3BP treatment, serum LDH level dropped maximally. Although a slow intravenous infusion of 3BP appeared to have minimal cytotoxicity, its anticancer efficacy via this delivery method was low. This was possibly due to high tumor GSH content, which was increased after concurrent use of the GSH depletor paracetamol. If the anticancer effectiveness of 3BP is less than expected, the combination with paracetamol may be needed to sensitize cancer cells to 3BP-induced effects.


Subject(s)
Acetaminophen , Therapeutic Uses , Adult , Carcinoma, Hepatocellular , Disease Progression , Drug Therapy, Combination , Enzyme Inhibitors , Glutathione , Glycolysis , Hexokinase , Humans , L-Lactate Dehydrogenase , Lactic Acid , Lung Neoplasms , Male , Melanoma , Drug Therapy , Necrosis , Neovascularization, Pathologic , Pleural Neoplasms , Prognosis , Pyruvates , Therapeutic Uses , Treatment Outcome
7.
Chinese Journal of Surgery ; (12): 450-454, 2014.
Article in Chinese | WPRIM | ID: wpr-314683

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of oral fluid resuscitation with pyruvate sodium-glucose-electrolyte solution (PGES) on hemodynamics, organ functions and mortalities during shock stage in dogs with burn.</p><p><b>METHODS</b>In comparison of oral pyruvate sodium-glucose-electrolyte solution (PGES) with NaHCO₃-glucose-electrolyte solution (HGES), beagle dogs with intubation of the carotid artery, jugular vein and jejunum for 24 hours were subjected to a 50% total body surface area (TBSA) burn, and were divided into three groups: pure burn without fluid resuscitation (NR, n = 8), and two oral fluid resuscitation (each n = 10), in which dogs were given with Pry-GES (OP) or NaHCO₃-GES (OH) according to Parkland formula. The hemodynamic and organ functions were measured serially before burn and 2, 6, 8, 12 and 24 hours after burn at no anaesthesia state A. Twenty-four hours mortality rate following burn was also recorded.</p><p><b>RESULTS</b>Two hours after burn, the mean arterial pressure of NR, OH and OP group was (45 ± 8), (57 ± 8) and (80 ± 9) mmHg (1 mmHg = 0.133 kPa) respectively, which were significantly reduced (t = 16.967, 14.595 and 10.100, all P < 0.05) compared with those before injury ((42 ± 6), (144 ± 6) and (142 ± 6) mmHg respectively), the change of cardiac output, dp/dtmax of left ventricular contractility and intestinal mucosal blood flow had the same trend as the mean arterial pressure. The systemic vascular resistance and organ parameters (Cr, CK-MB, ALT and DAO) in all groups increased obviously (t = -46.894--2.465, all P < 0.05). All measurements of NR group kept worsening, and all died within 24 hours after burn; while those of two oral resuscitation groups had improved gradually (F = 0.001-1.600, all P < 0.05), OP group was significantly superior to OH group (F = 0.013-0.466, P < 0.05). At 24 hours after burn, 6 (6/10) survived in OP group, 4 (4/10) in OH group and 0 (0/8) in NR group.</p><p><b>CONCLUSION</b>The Pyr-GES may be superior to the standard NaHCO₃-GES in the improvement of hemodynamics and organ functions during oral resuscitation in dogs with 50%TBSA full thickness burn.</p>


Subject(s)
Animals , Body Surface Area , Burns , Therapeutics , Disease Models, Animal , Dogs , Fluid Therapy , Hemodynamics , Male , Pyruvates , Therapeutic Uses , Rehydration Solutions , Shock , Therapeutics
8.
Article in Chinese | WPRIM | ID: wpr-356993

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of 3-bromopyruvate (3-BP) in sensitizing hepatocellular carcinoma cells to cisplatin-induced apoptosis and its possible mechanism.</p><p><b>METHODS</b>The growth inhibition of HepG2 and SMMC7721 cells following exposures to different concentrations of 3-BP and cisplatin was measured by MTT assay. The apoptosis of cells treated with 100 µmol/L 3-BP with or without 8 µmol/L cisplatin was assessed using flow cytometry with PI staining, and the activity of caspase-3 and intracellular ATP level were detected using commercial detection kits; the expression of XIAP and PARP was analyzed using Western blotting.</p><p><b>RESULTS</b>3-BP produced obvious inhibitory effects on HepG2 and SMMC7721 cells at the concentrations of 50-400 µmol/L with IC50 values of 238.9∓13.9 µmol/L and 278.7∓11.7 µmol/L for a 48-h treatment, respectively. Cisplatin also inhibited the growth of HepG2 and SMMC7721 cells at the concentrations of 2-32 µmol/L, with IC50 values of 16.4∓0.9 µmol/L and 20.9∓1.8 µmol/L after a 48-h treatment, respectively. Treatment with 100 µmol/L 3-BP combined with 8 µmol/L cisplatin for 48 h resulted in a growth inhibition rate of (60.6∓2.2)% in HepG2 cells and (56.8∓2.3)% in SMMC7721 cells, which were significantly higher than those in cells treated with 3-BP or cisplatin alone. The combined treatment for 48 h induced an apoptotic rate of (51.1∓4.3)% in HepG2 cells and (46.5∓3.9)% in SMMC7721 cells, which were also markedly higher than those in cells with 3-BP or cisplatin treatment alone.</p><p><b>CONCLUSION</b>3-BP can sensitize HepG2 and SMMC7721 cells to cisplatin-induced apoptosis possibly by causing intracellular ATP deficiency, down-regulating XIAP, and increasing caspase-3 activity.</p>


Subject(s)
Adenosine Triphosphate , Metabolism , Antineoplastic Agents , Pharmacology , Apoptosis , Carcinoma, Hepatocellular , Pathology , Caspase 3 , Metabolism , Cisplatin , Pharmacology , Hep G2 Cells , Humans , Liver Neoplasms , Pathology , Pyruvates , Pharmacology , X-Linked Inhibitor of Apoptosis Protein , Metabolism
9.
Acta cir. bras ; 28(8): 559-567, Aug. 2013. ilus, tab
Article in English | LILACS | ID: lil-680609

ABSTRACT

PURPOSE: To investigate if the ethyl-pyruvate solution could reduce mortality in AP and/or diminish the acute lung injury. METHODS: Forty male rats, weighing between 270 to 330 grams were operated. An experimental model of severe AP by injection of 0.1ml/100g of 2.5% sodium taurocholate into the bilio-pancreatic duct was utilized. The rats were divided into two groups of ten animals each: CT - control (treatment with 50ml/kg of Ringer's solution, intraperitoneal) and EP (treatment with 50ml/kg of Ringer ethyl- pyruvate solution, intra-peritoneal), three hours following AP induction. After six hours, a new infusion of the treatment solution was performed in each group. Two hours later, the animals were killed and the pulmonary parenchyma was resected for biomolecular analysis, consisting of: interleukin, myeloperoxidase, MDA, nitric oxide, metalloproteinases and heat shock protein. In the second part of the experiment, another, 20 rats were randomly divided into EP and CT groups, in order to evaluate a survival comparison between the two groups. RESULTS: There were no significant differences in IL-1B,IL-10, MMP-9, HSP70, nitric oxide, MPO, MDA (lipidic peroxidation) concerning both groups. The levels of IL-6 were significantly diminished in the EP group. Furthermore, the MMP-2 levels were also reduced in the EP group (p<0.05). The animals from the EP treatment groups had improved survival, when compared to control group (p<0.05). CONCLUSION: The ethyl-pyruvate diminishes acute lung injury inflammatory response in acute pancreatitis and ameliorates survival when compared to control group, in the experimental model of necrotizing acute pancreatitis.


Subject(s)
Animals , Male , Rats , Acute Lung Injury/drug therapy , Cytokines/metabolism , Matrix Metalloproteinases/metabolism , Pancreatitis, Acute Necrotizing/drug therapy , Pyruvates/pharmacology , Acute Lung Injury/chemically induced , Acute Lung Injury/enzymology , Disease Models, Animal , Immunoblotting , Isotonic Solutions/pharmacology , Kaplan-Meier Estimate , Pancreatitis, Acute Necrotizing/mortality , Random Allocation , Rats, Wistar , Reference Values , Reproducibility of Results , Time Factors , Treatment Outcome
10.
The Korean Journal of Pain ; : 135-141, 2013.
Article in English | WPRIM | ID: wpr-31288

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 , Apoptosis , Caspase 3 , Compliance , Diagnosis-Related Groups , Ganglia, Spinal , Glial Fibrillary Acidic Protein , Hyperalgesia , Myelin Sheath , Neuralgia , Neurons , Paclitaxel , Pyruvates , Pyruvic Acid , Rats
11.
Article in Chinese | WPRIM | ID: wpr-319424

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of glycolysis inhibitor 3-bromopyruvate (3-BrPA) in inducing apoptosis of human breast carcinoma cells SK-BR-3 and the possible mechanism.</p><p><b>METHODS</b>MTT assay was used to detect the growth inhibition induced by 3-BrPA in breast cancer cells SK-BR-3. The apoptotic cells were detected by flow cytometry with propidium iodide (PI). ATP levels in the cells were detected by ATP assay kit, and DHE fluorescent probe technique was used to determine superoxide anion levels; the mitochondrial membrane potential was assessed using JC-1 staining assay.</p><p><b>RESULTS</b>MTT assay showed that the proliferation of SK-BR-3 cells was inhibited by 3-BrPA in a time- and concentration-dependent manner. Exposure to 80, 160, and 320 µmol·L(-1) 3-BrPA for 24 h resulted in cell apoptosis rates of 6.7%, 22.3%, and 79.6%, respectively, and the intracellular ATP levels of SK-BR-3 cells treated with 80, 160, 320 µmol·L(-1) 3-BrPA for 5 h were 87.7%, 60.6%, and 23.7% of the control levels. 3-BrPA at 160 µmol·L(-1) increased reactive oxygen levels and lowered mitochondrial membrane potential of SK-BR-3 cells.</p><p><b>CONCLUSION</b>3-BrPA can inhibit cell proliferation, reduce the mitochondrial membrane potential and induce apoptosis in SK-BR-3 cells, the mechanism of which may involve a reduced ATP level by inhibiting glycolysis and increasing the reactive oxygen level in the cells.</p>


Subject(s)
Apoptosis , Cell Line, Tumor , Female , Glycolysis , Humans , Membrane Potential, Mitochondrial , Pyruvates , Pharmacology , Reactive Oxygen Species , Metabolism
12.
The Korean Journal of Pain ; : 213-220, 2012.
Article in English | WPRIM | ID: wpr-74032

ABSTRACT

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 , Apoptosis , Caspase 3 , Cell Death , Cold Temperature , Constriction , Diagnosis-Related Groups , Enzyme-Linked Immunosorbent Assay , Ganglia, Spinal , Hyperalgesia , Ligation , Neurons , Pyruvates , Pyruvic Acid , Rats , Spinal Nerve Roots , Spinal Nerves , Tumor Necrosis Factor-alpha
13.
Yonsei Medical Journal ; : 838-844, 2010.
Article in English | WPRIM | ID: wpr-72898

ABSTRACT

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 , Anti-Inflammatory Agents/pharmacology , Cell Nucleus/metabolism , Cytoplasm/metabolism , Heart/physiopathology , Inflammation , Male , Myocardial Infarction/prevention & control , Myocardium/metabolism , NF-kappa B/metabolism , Peroxidase/metabolism , Pyruvates/pharmacology , Rats , Rats, Sprague-Dawley , Reperfusion Injury/drug therapy
14.
Chinese Journal of Burns ; (6): 104-108, 2010.
Article in Chinese | WPRIM | ID: wpr-305617

ABSTRACT

<p><b>OBJECTIVE</b>To observe the influence of high mobility group box-1 protein (HMGB1) derived from spleen on the phenotype of regulatory T lymphocytes (Treg) and HMGB1-mediated immune function in severely scalded rats after delayed resuscitation.</p><p><b>METHODS</b>One hundred and four Wistar rats were divided into normal control group (NC, n = 8), sham scald group (SS, n = 32), scald group (S, n = 32), and ethyl pyruvate (EP) treatment group (EPT, n = 32) according to the random comparison table. Rats in the latter 2 groups were subjected to 30%TBSA full-thickness scald, which were intraperitoneally injected with Ringer solution or EP solution at post scald hour (PSH) 6 (delayed antishock treatment) and administered with 4 mL Ringer solution or EP solution per 12 hours after PSH 12 till PSH 48. Rats in SS group were treated the same as that of S group except for sham scald with 37 degrees C water. Injured rats were sacrificed at post scald day (PSD) 1, 3, 5, 7 (rats in NC group were also sacrificed), and CD4(+)CD25(+)Treg were isolated from spleen with magnetic-activated cell sorting method. The content of HMGB1 in spleen and IL-2 level in supernatant were determined with ELISA. The expression of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) on Treg was determined with flow cytometry, and the proliferation activity of T lymphocytes was also detected (recorded as absorbance value). Data were processed with analysis of variance among groups and independent samples t test.</p><p><b>RESULTS</b>(1) Compared with that of rats in SS group and EPT group, the expression of splenic HMGB1 in S group increased significantly on PSD 1 through PSD 7 [peaked on PSD 1: (46.7 +/- 8.3) ng/mg protein]. (2) Compared with that in SS group, the expression of CTLA-4 in S group was enhanced significantly on PSD 1 through PSD 5 (with t value respectively 10.459, 12.051, 4.029, P < 0.05 or P < 0.01); while that in EPT group decreased significantly on PSD 1 through PSD 7 as compared with that from S group (with t value respectively 2.796, 9.913, 9.581, 10.022, P < 0.05 or P < 0.01). (3) Compared with that of rats in SS group, the proliferation activity of T lymphocytes in S group was markedly suppressed on PSD 1 through PSD 7 (nadir on PSD1: 0.167 +/- 0.059), and release of IL-2 was decreased significantly [nadir on PSD 5: (44 +/- 24) pg/mL]. T lymphocytes proliferation activity was restored and excretion of IL-2 increased in EPT group as compared respectively with that of S group at each time point.</p><p><b>CONCLUSIONS</b>The release of HMGB1 may stimulate splenic Treg to mature, thereby induce suppression of proliferation activity of T lymphocytes and immune function. EP can ameliorate immune dysfunction in animals with delayed resuscitation through inhibiting the synthesis and release of HMGB1.</p>


Subject(s)
Animals , Antigens, CD , Metabolism , Burns , Allergy and Immunology , CTLA-4 Antigen , Cell Proliferation , HMGB1 Protein , Metabolism , Interleukin-2 , Metabolism , Male , Pyruvates , Pharmacology , Rats , Rats, Wistar , Spleen , Cell Biology , Allergy and Immunology , T-Lymphocytes, Regulatory , Cell Biology , Allergy and Immunology
15.
Chinese Medical Journal ; (24): 1681-1685, 2009.
Article in English | WPRIM | ID: wpr-240844

ABSTRACT

<p><b>BACKGROUND</b>Breast cancer is one of the most common malignancies in women and is highly resistant to chemotherapy. Due to its high tumour selectivity, 3-bromopyruvic acid (3-BrPA), a well-known inhibitor of energy metabolism has been proposed as a specific anticancer agent. The present study determined the effect of 3-BrPA on proliferation, cell cycle and apoptosis in the human breast cancer MCF-7 cell line and other antitumour mechanisms.</p><p><b>METHODS</b>MCF-7 cells were treated with various concentrations of 3-BrPA for 1 - 4 days, and cell growth was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. Marked morphological changes in MCF-7 cells after treatment with 3-BrPA were observed using transmission electron microscopy. The distributions of the cell cycle and apoptosis were analyzed by flow cytometry. Immunohistochemistry was used to indicate the changes in the expression of Bcl-2, c-Myc, and mutant p53.</p><p><b>RESULTS</b>3-BrPA (25 microg/ml) significantly inhibited the proliferation of MCF-7 cells in a time-dependent manner. The MCF-7 cells exposed to 3-BrPA showed the typical morphological characteristics of apoptosis, including karyopycnosis, nuclear condensation and oversize cytoplasmic particles. In addition, flow cytometric assay also showed more apoptotic cells after 3-BrPA stimulation. The cells at the G0 and G1 phases were dramatically decreased while cells at the S and G2/M phases were increased in response to 3-BrPA treatment after 48 hours. Furthermore, 3-BrPA stimulation decreased the expressions of Bcl-2, c-Myc and mutant p53, which were strongly associated with the programmed cell death signal transduction pathway.</p><p><b>CONCLUSION</b>3-BrPA inhibits proliferation, induces S phase and G2/M phase arrest, and promotes apoptosis in MCF-7 cells, which processes might be mediated by the downregulation of the expressions of Bcl-2, c-Myc and mutant p53.</p>


Subject(s)
Antineoplastic Agents , Chemistry , Pharmacology , Apoptosis , Breast Neoplasms , Cell Cycle , Cell Division , Cell Line, Tumor , Cell Proliferation , Female , Flow Cytometry , G2 Phase , Humans , Immunohistochemistry , Molecular Structure , Pyruvates , Chemistry , Pharmacology , S Phase
16.
Article in English | WPRIM | ID: wpr-123977

ABSTRACT

OBJECTIVE: The purpose of this study was to compare the antitumor effect and hepatotoxicity of an intraarterial delivery of low-dose and high-dose 3-bromopyruvate (3-BrPA) and those of a conventional Lipiodol-doxorubicin emulsion in a rabbit VX2 hepatoma model. MATERIALS AND METHODS: This experiment was approved by the animal care committee at our institution. VX2 carcinoma was implanted in the livers of 36 rabbits. Transcatheter intraarterial administration was performed using low dose 3-BrPA (25 mL in a 1 mM concentration, n = 10), high dose 3-BrPA (25 mL in a 5 mM concentration, n = 10) and Lipiodol-doxorubicin emulsion (1.6 mg doxorubicin/ 0.4 mL Lipiodol, n = 10), and six rabbits were treated with normal saline alone as a control group. One week later, the proportion of tumor necrosis was calculated based on histopathologic examination. The hepatotoxicity was evaluated by biochemical analysis. The differences between these groups were statistically assessed with using Mann-Whitney U tests and Kruskal-Wallis tests. RESULTS: The tumor necrosis rate was significantly higher in the high dose group (93% +/- 7.6 [mean +/- SD]) than that in the control group (48% +/- 21.7) (p = 0.0002), but the tumor necrosis rate was not significantly higher in the low dose group (62% +/- 20.0) (p = 0.2780). However, the tumor necrosis rate of the high dose group was significantly lower than that of the Lipiodol-doxorubicin treatment group (99% +/- 2.7) (p = 0.0015). The hepatotoxicity observed in the 3-BrPA groups was comparable to that of the Lipiodol-doxorubicin group. CONCLUSION: Even though intraarterial delivery of 3-BrPA shows a dose-related antitumor effect, single session treatment seems to have limited efficacy when compared with the conventional method.


Subject(s)
Animals , Disease Models, Animal , Dose-Response Relationship, Drug , Doxorubicin/administration & dosage , Infusions, Intra-Arterial , Iodized Oil/administration & dosage , Liver Neoplasms, Experimental/drug therapy , Pyruvates/administration & dosage , Rabbits , Statistics, Nonparametric , Tomography, X-Ray Computed
17.
Article in English | WPRIM | ID: wpr-634605

ABSTRACT

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 120 min. In the I/R group, after 30 min stabilization the injury was induced by 30 min global ischemia followed by 60 min reperfusion. Ethyl pyruvate (EP) group was set up with the same protocol as I/R group except that it was supplied with 2 mmol/L EP 15 min 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.


Subject(s)
Apoptosis , In Situ Nick-End Labeling , Malondialdehyde/pharmacology , Myocardium/pathology , Myocytes, Cardiac/cytology , Oxidative Stress , Proto-Oncogene Proteins c-bcl-2/metabolism , Pyruvates/pharmacology , Rats, Sprague-Dawley , Reperfusion Injury , Tissue Distribution , bcl-2-Associated X Protein/metabolism
18.
Article in Chinese | WPRIM | ID: wpr-273867

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of ethyl pyruvate on barrier function of intestinal mucosa in dogs with septic shock.</p><p><b>METHODS</b>Twenty dogs with septic shock induced by lipopolysaccharides(LPS) were randomly divided into two groups. Dogs randomly received placebo (Ringer's solution, control group, n=8) or ethyl pyruvate in lactated Ringer's solution (0.05 g/kg loading dose over 10 mins, thereafter 0.05 g.kg(-1).h(-1) for 12 hours, EP treatment group, n=12). The diamine oxidase(DAO) activity and D-lactate content were detected at the 0, 8 th, 12 th and 24 th hour of septic shock. Animals were sacrificed at the 24 th hour after septic shock and the jejunal tissue was taken for histopathological examination.</p><p><b>RESULTS</b>The levels of plasma DAO and D-lactate were significantly elevated in both groups after septic shock than those before septic shock. The changes in intestinal parameters of hemoperfusion and permeability in EP treatment group were significantly lowered than those in control group. Inflammation of small intestinal mucosa was more severe in control group than that in EP group, and the pathologic score was significantly lower in EP group(2.33+/-0.25) than that in control group(3.39+/-0.38)(P<0.05).</p><p><b>CONCLUSION</b>Ethyl pyruvate can lessen intestinal permeability and protect intestinal barrier function in dogs with septic shock.</p>


Subject(s)
Animals , Dogs , Intestinal Mucosa , Pathology , Intestine, Small , Male , Pyruvates , Therapeutic Uses , Shock, Septic , Drug Therapy , Pathology
19.
Article in English | WPRIM | ID: wpr-62114

ABSTRACT

OBJECTIVE: We wanted to investigate the feasibility of using FDG-PET for evaluating the antitumor effect of intraarterial administration of a hexokinase II inhibitor, 3-bromopyruvate (3-BrPA), in a rabbit VX2 liver tumor model. MATERIALS AND METHODS: VX2 carcinoma was grown in the livers of ten rabbits. Two weeks later, liver CT was performed to confirm appropriate tumor growth for the experiment. After tumor volume-matched grouping of the rabbits, transcatheter intraarterial administration of 3-BrPA was performed (1 mM and 5 mM in five animals each, respectively). FDG-PET scan was performed the day before, immediately after and a week after 3-BrPA administration. FDG uptake was semiquantified by measuring the standardized uptake value (SUV). A week after treatment, the experimental animals were sacrificed and the necrosis rates of the tumors were calculated based on the histopathology. RESULTS: The SUV of the VX2 tumors before treatment (3.87+/-1.51[mean+/-SD]) was significantly higher than that of nontumorous liver parenchyma (1.72+/-0.34) (p < 0.0001, Mann-Whitney U test). The SUV was significantly decreased immediately after 3-BrPA administration (2.05+/-1.21) (p = 0.002, Wilcoxon signed rank test). On the one-week follow up PET scan, the FDG uptake remained significantly lower (SUV 1.41+/-0.73) than that before treatment (p = 0.002), although three out of ten animals showed a slightly increasing tendency for the FDG uptake. The tumor necrosis rate ranged from 50.00% to 99.90% (85.48%+/-15.87). There was no significant correlation between the SUV or the SUV decrease rate and the tumor necrosis rate in that range. CONCLUSION: Even though FDG-PET cannot exactly reflect the tumor necrosis rate, FDG-PET is a useful modality for the early assessment of the antitumor effect of intraarterial administration of 3-BrPA in VX2 liver tumor.


Subject(s)
Animals , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Feasibility Studies , Fluorodeoxyglucose F18 , Infusions, Intra-Arterial , Injections, Intra-Arterial , Liver Neoplasms, Experimental/drug therapy , Necrosis , Positron-Emission Tomography , Pyruvate Dehydrogenase Complex/antagonists & inhibitors , Pyruvates/pharmacology , Rabbits , Radiopharmaceuticals
20.
Chinese Journal of Surgery ; (12): 1210-1213, 2007.
Article in Chinese | WPRIM | ID: wpr-340827

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of ethyl pyruvate (EP) on high mobility group box-1 protein (HMGB1) expression in renal tissue and acute kidney injury in rats with delayed resuscitation after thermal injury.</p><p><b>METHODS</b>Seventy-eight Wistar rats subjected to 30% total body surface area full-thickness thermal injury followed with delayed resuscitation were divided into 3 groups: sham group (n = 18), injury group (n = 30) and EP group (n = 30). Renal tissue and blood samples were harvested to determine HMGB1 mRNA as well as its protein expression and renal function parameter at the 8, 24, 72 h post the "injury". HMGB1 mRNA was semi-quantitatively measured by reverse transcription polymerase chain reaction taking GAPDH as an internal standard, and HMGB1 protein expression was determined by Western blot and immunohistochemistry. Blood urea nitrogen (BUN) levels were measured with automatic biochemistry analyzer. The pathological changes of renal tissues were examined using HE staining.</p><p><b>RESULTS</b>Compared with sham controls, both mRNA and protein expressions of HMGB1 in injury group were significantly enhanced in kidneys at 8 - 72 h after thermal injury (P < 0.05), meanwhile serum BUN levels were markedly increased (P < 0.05). Compared with injury group, the renal HMGB1 mRNA and protein expressions were markedly down-regulated in EP group at 8 h, 24 h and 72 h post injury (P < 0.05), respectively, and meanwhile serum BUN levels were reduced significantly (P < 0.05). Inflammatory cell infiltration was found in renal tissues following injury, and kidney injury was markedly alleviated after treatment with EP.</p><p><b>CONCLUSIONS</b>It indicated that HMGB1 appears to be involved in the pathogenesis of post-burn acute kidney injury. Treatment with EP reduces renal HMGB1 expression, and protects against acute kidney injury secondary to delayed resuscitation after major burns.</p>


Subject(s)
Acute Disease , Animals , Blood Urea Nitrogen , Blotting, Western , Burns , Blood , Therapeutics , Disease Models, Animal , HMGB1 Protein , Genetics , Metabolism , Immunohistochemistry , Kidney , Metabolism , Pathology , Kidney Diseases , Genetics , Metabolism , Male , Pyruvates , Pharmacology , RNA, Messenger , Genetics , Metabolism , Random Allocation , Rats , Rats, Wistar , Resuscitation , Reverse Transcriptase Polymerase Chain Reaction , Time Factors
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