The role of TNF-α and NFkß in an experimental model of intestinal carcinogenesis with 1, 2-dimethyhydrazine

ABSTRACT Purpose: To analyze the potential of tumor necrosis factor-α (TNF-α) and factor nuclear kappa B (NF-κB) as colorectal cancer (CRC) biomarkers in an experimental model of intestinal carcinogenesis with 1,2-dimethyhydrazine (1,2-DMH). Methods: Twenty-four male Wistar rats were divided into two groups: sham and 1,2-DMH. First, 1,2-DMH (20 mg/kg/week) was administered for 15 consecutive weeks. In the 25th week, proctocolectomy was conducted. Histopathological analysis, immunohistochemistry, and gene expression of TNF-α and NF-κB were performed. Statistical analysis was performed using GraphPad Prism. The location of aberrant crypt foci (ACF) was analyzed by Kruskal-Wallis' test. For analyses with two groups with parametric data, the t-test was used; for non-parametric data, the Mann-Whitney's test was used. P < 0.05 was considered significant. Results: The number of ACF and macroscopic lesions was significantly higher (p < 0.5) in the 1,2-DMH group compared to the sham group, and most ACF were concentrated in the distal segment of the colon. There was a statistically significant increase (p < 0.5) in protein and gene expression of TNF-α and NF-κB in the 1,2-DMH group compared to the sham group. Conclusions: Our results provide supportive evidence that TNF-α and NF-κB pathways are strongly involved in CRC development in rats and might be used as early biomarkers of CRC pathogenesis in experimental studies.

Preconditioning with L-Ala-Gln reduces the expression of inflammatory markers (TNF-α, NF-κB, IL-6 and HO-1) in an injury animal model of cerebrovascular ischemia in Meriones unguiculatus (gerbils)

Abstract Purpose To evaluate the neuroprotective effect of L-alanyl-glutamine in a gerbil model of brain ischemia-reperfusion injury based on immunohistochemical quantification of pro-inflammatory and cell activation biomarkers (TNF-α, NF-κB, IL-6 and HO-1). Methods Male gerbils weighing 100-180 g were pretreated with either 0.75 g/kg L-Ala-Gln (n=18) or 2.0 mL saline (n=18) administered i.v. 30 minutes before the bilateral ligation of the common carotid artery during 15 min and then the ligation was removed. Under anesthesia with urethane, brain tissue was harvested at 0 min (T0), 30 min (T30) and 60 min (T60) after reperfusion. The tissue was embedded in 10% formalin overnight and 4-μm sections were prepared for immunostaining with monoclonal antibodies. Immunostained cells were counted by optical microscopy. The statistical analysis used mean values based on 4 sections. Results The pretreatment with L-Ala-Gln animal group 1 demonstrated significantly lower levels of TNF-α, NF-κB and IL-6. On the other hand, the levels of HO-1 were significantly higher, suggesting a protective role in model of brain ischemia-reperfusion injury. Conclusion These findings suggest a protective effect of L-Ala-Gln by decreasing levels of TNF-alpha, IL-6 and NF-κB and Increasing levels of HO-1.

Comparative study on the efficacy of non-steroidal, steroid and non-use of anti-inflammatory in the treatment of acute epidemic conjunctivitis

Abstract Purpose To evaluate the effects of prednisolone against sodium diclofenac both with ciprofloxacin compared to artificial tears on the symptoms and signs of acute viral conjunctivitis. Methods Study included 37 patients diagnosed with acute conjunctivitis and distributed by three groups: A (1% prednisolone acetate + ciprofloxacin (0.3%); B (Sodium diclofenac (0.1%) + ciprofloxacin (0.3%) and C (artificial tears + ciprofloxacin (0.3%). Patients received medication 6/6 hours daily. Signs and symptoms (e.g. lacrimation, burning, photophobia, etc.) were scored at baseline and on the first, third, fifth and seventh days and in the end of treatment using a standardized questionnaire and slit lamp anterior segment examination. Results All three groups demonstrated an improvement in the signs and symptoms of conjunctivitis in their follow-up visits. There was no significant difference in symptom and sign scores between Group A and B and B and C in the study visits ( p >0.05). However, the comparison between groups A and C showed a clinical trend (p=0.05) on third evaluation suggesting better clinical action using the corticosteroids. Conclusion The prednisolone acetate was not superior to the use of sodium diclofenac or artificial tears in relieving the signs and symptoms of viral conjunctivitis.

Association of oxidative stress and DNA damage with grafting time in patients with multiple myeloma and lymphoma submitted to autologous hematopoietic stem cell transplantation / Associação do estresse oxidativo e dano ao DNA com o tempo de enxertia em pacientes com mieloma múltiplo e linfomas submetidos a transplante autólogo de células-tronco hematopoéticas

ABSTRACT The aim of the study was to investigate the association between oxidative stress and DNA damage with grafting time in patients submitted to autologous hematopoietic stem-cell transplantation (HSCT). The study included 37 patients submitted to autologous HSCT diagnosed with Multiple Myeloma (MM) and lymphoma (Hodgkin’s and non-Hodgkin’s). Biomarkers of oxidative stress and DNA damage index (DI) were performed at baseline (pre-CR) of the disease and during the conditioning regimen (CR), one day after the HSCT, ten days after HSCT and twenty days after HSCT, as well as in the control group consisting of 30 healthy individuals. The outcomes showed that both groups of patients had an hyperoxidative state with high DI when compared to baseline and to the control group and that the CR exacerbated this condition. However, after the follow-up period of the study, this picture was re-established to the baseline levels of each pathology. The study patients with MM showed a mean grafting time of 10.75 days (8 to 13 days), with 10.15 days (8 to 15 days) for the lymphoma patients. In patients with MM, there was a negative correlation between the grafting time and the basal levels of GPx (r = -0.54; p = 0.034), indicating that lower levels of this important enzyme are associated with a longer grafting time. For the DI, the correlation was a positive one (r = 0.529; p = 0.030). In the group with lymphoma, it was observed that the basal levels of NOx were positively correlated with grafting time (r = 0.4664, p = 0.032). The data indicate the potential of these biomarkers as predictors of toxicity and grafting time in patients with MM and Lymphomas submitted to autologous HSCT.

RESUMO O objetivo do estudo foi investigar a associação entre estresse oxidativo e dano ao DNA com o tempo de enxertia em pacientes submetidos ao transplante de células-tronco hematopoéticas autólogo (TCTH). Participaram do estudo 37 pacientes submetidos ao TCTH autólogo com diagnóstico de mieloma múltiplo (MM) e Linfomas (Hodgkin e não Hodgkin). Biomarcadores de estresse oxidativo e índice de dano ao DNA (ID) foram determinados no estado basal (Pré-RC) das doenças e durante o regime de condicionamento (RC), um dia após o TCTH, dez dias após o TCTH e vinte dias após o TCTH e no grupo controle composto por 30 individuos saudáveis. Os resultados demonstraram que os dois grupos de pacientes apresentaram um estado hiperoxidativo com elevado ID quando comparados ao estado basal e ao grupo controle e que o RC exacerbou essa condição. No entanto, após o tempo de acompanhamento do estudo, esse quadro foi reestabelecido ao estado basal de cada patologia. Os pacientes do estudo com MM apresentaram uma média do tempo de enxertia de 10,75 dias (8 a 13 dias), e de 10,15 dias (8 a 15 dias) para o grupo Linfoma. Nos pacientes com MM houve uma correlação negativa entre o tempo de enxertia e os níveis basais de GPx (r=-0,54; p=0,034), indicando que níveis mais baixos de GPx estão relacionados a um maior tempo de enxertia, e para o ID, a correlação foi positiva (r=0,529; p=0,030). No grupo com Linfoma, observou-se que os níveis basais de NOx correlacionaram-se positivamente com o tempo de enxertia (r= 0,4664; p=0,032). Os dados apontam para o potencial desses biomarcadores como preditores da toxicidade e do tempo de enxertia em pacientes com MM e Linfomas submetidos ao TCTH autólogo

Relevance of prognostic factors in the decision-making of stem cell transplantation in Myelodysplastic Syndromes / Relevância dos fatores prognósticos na decisão do transplante de células-tronco na Síndrome Mielodisplásica

ABSTRACT The hematopoietic stem cell transplantation (HSCT) is the only curative alternative for Myelodysplastic Syndrome (MDS), but many patients are not eligible for this treatment, as there are several limiting factors, especially in the case of patients with low-risk MDS. The aim of this study is to discuss the factors that can guide the decision-making on referring or not a patient to HSCT. Three cases of MDS, two of which were submitted to HSCT are presented. We intend to report the difficulties in referring patients with MDS to transplant and the prognostic factors that contribute to define eligibility.

RESUMO O transplante de células-tronco hematopoéticas (TCTH) é a única alternativa curativa para Síndrome Mielodisplásica (SMD), porém muitos pacientes não são elegíveis para esta opção, pois existem diversos fatores limitantes, principalmente no caso de pacientes com SMD de baixo risco. O objetivo do estudo é discutir os fatores que podem orientar a decisão no encaminhamento ou não para o TCTH. São apresentados três casos de SMD, dos quais dois foram submetidos ao TCTH. Nos propomos a relatar as dificuldades no encaminhamento dos pacientes com SMD ao transplante e os fatores prognósticos que contribuem para definir a elegibilidade.

Oil mixes omega 9, 6 and 3, enriched with seaweed, promoted reduction of thermal burned modulating NF-kB and Ki-67

PURPOSE: To examine the effects of the oil mixes (ω-9, ω-6 and ω-3) in rats subjected to thermal burn. It was also aimed to assess whether the sources of ω3 would interfere with the effect of such mixes on the thermal injury. METHODS: Thirty-six rats distributed into five groups: burned + water, burned + isolipid mix, burned + oil mix 1 (ALA), burned + oil mix 2 (ALA + EPA + DHA of fish) and burned + oil mix 3 (ALA + DHA from seaweed). The thermal injury was involving total thickness of skin. After the burns animals received the oil mixes for seven days. The lesions were evaluated by immunohistochemistry. RESULTS: Animals receiving mix 3 showed a smaller extension of the thermal injury as compared to those that were supplemented with other oils mixes. Expression of Ki-67 in the receiving Mix 3 increased as compared to all the other groups. Animals supplemented with mix 3 were able to inhibit NF-κB in injured tissue. CONCLUSION: Rats received oil mix in which the source of ω3 (ALA+DHA of seaweed) showed inhibition of NF-κB, increase in cell proliferation, and reduction the extension of thermal lesion. .

Fructo-oligosaccharide effects on serum cholesterol levels. An overview

PURPOSE: To address the effects of fructooligosaccharides (FOS) intake on serum cholesterol levels. METHODS: We performed a search for scientific articles in MEDLINE database from 1987 to 2014, using the following English keywords: fructooligosaccharides; fructooligosaccharides and cholesterol. A total of 493 articles were found. After careful selection and exclusion of duplicate articles 34 references were selected. Revised texts were divided into two topics: "FOS Metabolism" and "FOS effects on plasma cholesterol." RESULTS: The use of a FOS diet prevented some lipid disorders and lowered fatty acid synthase activity in the liver in insulin-resistant rats. There was also reduction in weight and total cholesterol in beagle dogs on a calorie-restricted diet enriched with short-chain FOS. Another study found that 2g FOS daily consumption increased significantly serum HDL cholesterol levels but did not ensure a significant reduction in levels of total cholesterol and triglycerides.. Patients with mild hypercholesterolemia receiving short-chain FOS 10.6g daily presented no statistically significant reduction in serum cholesterol levels. However, when FOS was offered to patients that changed their lifestyle, the reduction of LDL cholesterol and steatosis was higher. CONCLUSIONS: Fructooligosaccharides intake may have a beneficial effect on lipid metabolism and regulation of serum cholesterol levels in individuals that change their lifestyle. FOS supplementation use in diets may therefore be a strategy for lowering cholesterol. .

Preconditioning with L-alanyl-glutamine upon cerebral edema and hypocampus red neurons counting in rats subjected to brain ischemia /reperfusion injury

PURPOSE: To evaluate the effects of the dipeptide L-alanyl-glutamine (L-Ala-Gln) as a preconditioning agent to potentially promote reduction in the intensity of lesion or induction of resilience in rats subjected to global cerebral ischemia/reperfusion (I/R) injury. METHODS: Thirty-six male Wistar rats weighing 280-300g were randomly assigned to six groups (n=6). Groups Sham 1h and 24h were treated with saline and spared of further interventions. The remaining groups were submitted to clamping of the common carotid arteries for 30 minutes (ischemia) and treated with saline (SS) or L-Ala-Gln. Brain reperfusion was allowed for 1or 24 h. L-Ala-Gln was administered intravenously (0.75g/kg) 30 minutes before sham procedure or induction of global brain I/R injury. Brain edema and red neuron counting were determined. Results were expressed as Mean±SD for normal results and Median±Percentile for non parametric data. Significance was established at p<0.05. RESULTS: Global I/R injury promoted an increase in brain edema at 24 h after reperfusion, whereas preconditioning with L-Ala-Gln induced no change in edema. On the other hand, L-Ala-Gln preconditioning decreased significantly red neurons counting both at 1h and 24h post reperfusion (p<0.05). CONCLUSION: There was a significant preconditioning effect with L-Ala-Gln decreasing cell death (red neurons counting) at early (1h) and late reperfusion (24h) in the cerebral tissue. .

Preconditioning with mono and polyunsaturated fatty acids and low-intensity electrical stimulation. Effects on skin repair in rats

PURPOSE: To evaluate the effects of preconditioning with oils mixes containing ω3/ω6/ω9 associated with micro-currents on skin repair in rats. METHODS: One-hundred and eight Wistar rats randomized into G-1, G-2 and G-3 groups were treated with saline (0.9%), mix 1 (corn+soybean oils) and mix 2 (olive+canola+flaxseed oils), respectively, in a single dose (0.01ml/g) by gavage. Next, each group was subdivided into sham and stimulated subgroups. Pulsed-wave microcurrents (0.5 µA, 0.5 Hz) were applied to stimulated subgroups for 20 min. One hour later anesthetized rats were subjected to surgery. A dorsal incision (6 cm long) was carried out and closed with interrupted nylon sutures. Samples (1cm2 ) were harvested from the mid-portion of the incision on the 7, 14, 21 post-operative (P.O.) days. Variables were analyzed using Mann-Whitney/Dunn tests Significance level was set to 5 % (p<0.05). RESULTS: Micro-currents promoted increase of exudate and reduction of epithelialization on day 7 in G1 rats. Mixes 1/2 reduced vascularization on 7/14th days P.O. Both 1/2 mixes reduced fibrosis on day 14. Preconditioning with mix 1 led to increased expression of NF-kB on the 7th day. CONCLUSION: Preconditioning with microcurrents has pro-inflammatory effects while oil mixes 1 and 2 decrease fibrosis and vascularization in the proliferative phase of cicatrization.

Preconditioning with mono and polyunsaturated fatty acids and low-intensity electrical stimulation. Effects on skin repair in rats

PURPOSE: To evaluate the effects of preconditioning with oils mixes containing ω3/ω6/ω9 associated with micro-currents on skin repair in rats. METHODS: One-hundred and eight Wistar rats randomized into G-1, G-2 and G-3 groups were treated with saline (0.9%), mix 1 (corn+soybean oils) and mix 2 (olive+canola+flaxseed oils), respectively, in a single dose (0.01ml/g) by gavage. Next, each group was subdivided into sham and stimulated subgroups. Pulsed-wave microcurrents (0.5 µA, 0.5 Hz) were applied to stimulated subgroups for 20 min. One hour later anesthetized rats were subjected to surgery. A dorsal incision (6 cm long) was carried out and closed with interrupted nylon sutures. Samples (1cm2) were harvested from the mid-portion of the incision on the 7, 14, 21 post-operative (P.O.) days. Variables were analyzed using Mann-Whitney/Dunn tests Significance level was set to 5 % (p<0.05). RESULTS: Micro-currents promoted increase of exudate and reduction of epithelialization on day 7 in G1 rats. Mixes 1/2 reduced vascularization on 7/14th days P.O. Both 1/2 mixes reduced fibrosis on day 14. Preconditioning with mix 1 led to increased expression of NF-kB on the 7th day. CONCLUSION: Preconditioning with microcurrents has pro-inflammatory effects while oil mixes 1 and 2 decrease fibrosis and vascularization in the proliferative phase of cicatrization. .

Development of a subcutaneous endometriosis rat model

PURPOSE: To present a rat model of subcutaneous endometriosis for the study of pathophysiology and the effects of drugs. METHODS: Fifty three-month-old female Wistar rats (Rattus norvergicus) were distributed into one control group and four treatment groups: estradiol (2.5; 5; 10mg/kg sc), medroxyprogesterone acetate (0.5; 2; 5mg/kg sc), triptorelin pamoate (0.18; 0.56mg/kg sc) and acetylsalicylic acid (3mg/kg per os). The animals were autoimplanted subcutaneously with 4x4-mm uterine fragments to induce endometriosis. The endometriomas were measured on days 1, 7, 14 and 21. The relative dry and wet weights of the endometrioma were used to evaluate response to the drug. Endometrial -like tissue was confirmed by histology. The greatest weight gain was observed on day 14 (relative wet weight: 29.1 ± 6.7mg%, relative dry weight: 5.3 ± 0.9mg %). Treatments were administered between day 5 and day 14. RESULTS: The relative wet weight of the hemiuterus in the 10mg/kg estradiol group differed significantly from control and the other two estradiol groups (p=0.0001). In the medroxyprogesterone acetate group the weight decreased significantly but this decrease was not dose-dependent. Weight reduction was also significant in the triptorelin pamoate and the acetylsalicylic acid groups. CONCLUSION: The model of subcutaneous endometriosis is reproducible, low-cost and easy to perform, and suitable for the study of pathophysiology and the effects of drugs. .

Nutraceutical preconditioning with arginine and oil mixes. Effects on inflammatory mediators, oxidative stress and lipid profile in patients undergoing radical prostatectomy

PURPOSE: To investigate whether there is any effect resulting from preconditioning with nutraceutical supplementation containing arginine and oil mixes with high ω9:ω6 ratio and low ω6:ω3 ratio containing EPA and DHA, ALA fatty acids on inflammatory mediators, antioxidant and lipid profile modulation in surgical trauma. METHODS: Twenty-six men scheduled for radical prostatectomy were randomized into three groups and treated as follows: Group 1 (skim milk, 0% fat), Group 2 (supplement with ω6:ω3 ratio of 8:1 and arginine) and Group 3 (supplement with high ω9:ω6 ratio of 3.2:1 and low ω6:ω3 ratio of 1.4:1 and arginine). Patients received skin milk or supplements twice a day (200 ml) during five days prior to surgery. Peripheral venous blood samples were collected at three different timepoints: five days before surgery (PRE), before anesthesia induction (IND) and on the 2nd postoperative day (POS). Parameters analyzed included inflammatory cytokines (IL-1β, IL-6, IL-8 and TNF-α), antioxidants (catalase), lipid profile and heat shock protein (HSP-27). RESULTS: There were no significant differences between groups on inflammatory mediators and antioxidant parameters. However, lipid profile values (Cholesterol, LDL, Triglycerides, VLDL), were significantly different. CONCLUSION: Preconditioning with arginine and oil mixes containing high ω9:ω6 ratio and low ω6:ω3 ratio, has no effects on inflammatory mediators and oxidative stress in patients undergoing radical prostatectomy. Reduction of cholesterol, triglycerides, LDL and VLDL profiles may be related to the trauma effect. .

Tumor suppressor p53 protein expression: prognostic significance in patients with low-risk myelodysplastic syndrome

BACKGROUND: At the time of diagnosis, more than 50% of patients with myelodysplastic syndrome have a normal karyotype and are classified as having a favorable prognosis. However, these patients often show very variable clinical outcomes. Furthermore, current diagnostic tools lack the ability to look at genetic factors beyond karyotyping in order to determine the cause of this variability. OBJECTIVE: To evaluate the impact of p53 protein expression at diagnosis in patients with low-risk myelodysplastic syndrome. METHODS: This study enrolled 38 patients diagnosed with low-risk myelodysplastic syndrome. Clinical data were collected by reviewing medical records, and immunohistochemical p53 staining was performed on bone marrow biopsies. RESULTS: Of the 38 participants, 13 (34.21%) showed p53 expression in their bone marrow. At diagnosis, this group of patients also presented clinical features characteristic of a poor prognosis more often than patients who did not express p53. Furthermore, patients expressing p53 had a shorter median survival time compared to those without p53 expression. CONCLUSION: This study shows that the expression of p53 at diagnosis is a useful indicator of distinct clinical characteristics and laboratory profiles found in low-risk myelodysplastic syndrome patients. These data indicate that the immunohistochemical analysis of p53 may be a prognostic tool for myelodysplastic syndrome and should be used as an auxiliary test to help determine the best therapeutic choice. .

Glutamine and ornithine alpha-ketoglutarate supplementation on malate dehydrogenases expression in hepatectomized rats

PURPOSE: To evaluate the relative gene expression (RGE) of cytosolic (MDH1) and mitochondrial (MDH2) malate dehydrogenases enzymes in partially hepatectomized rats after glutamine (GLN) or ornithine alpha-ketoglutarate (OKG) suplementation. METHODS: One-hundred and eight male Wistar rats were randomly distributed into six groups (n=18): CCaL, GLNL and OKGL and fed calcium caseinate (CCa), GLN and OKG, 0.5g/Kg by gavage, 30 minutes before laparotomy. CCaH, GLNH and OKGH groups were likewise fed 30 minutes before 70% partial hepatectomy. Blood and liver samples were collected three, seven and 14 days after laparotomy/hepatectomy for quantification of MDH1/MDH2 enzymes using the real-time polymerase chain reaction (PCR) methodology. Relative enzymes expression was calculated by the 2-ΔΔC T method using the threshold cycle (CT) value for normalization. RESULTS: MDH1/MDH2 RGE was not different in hepatectomized rats treated with OKG compared to rats treated with CCa. However, MDH1/MDH2 RGE was greater on days 3 (321:1/26.48:1) and 7 (2.12:1/2.48:1) while MDH2 RGE was greater on day 14 (7.79:1) in hepatectomized rats treated with GLN compared to control animals. CONCLUSION: Glutamine has beneficial effects in liver regeneration in rats by promoting an up-regulation of the MDH1 and MDH2 relative gene expression. .

Ternatin pretreatment attenuates testicular injury induced by torsion/detorsion in Wistar rats / Pré-tratamento com ternatina atenua a lesão testicular induzida por torção/destorção em ratos Wistar

PURPOSE: To investigate the possible protective role of the bioflavonoid ternatin (TTN) when administered before induction of ischemia/reperfusion injury in rat testis. METHODS: Thirty-six Wistar rats were randomly assigned to 3 groups (n=12), divided in 2 subgroups (n=6). Saline 2.0ml (G-1), dimethylsulfoxide (DMSO) 3 percent solution (G-2) or TTN 12 mg/kg/dose (G-3) was administered ip. to all rats, respectively, 21, 12 and 1 hour before torsion. Anesthetized rats were subjected to ischemia (3 hours) induced by 720º torsion of the spermatic cord. Right testis and arterial blood samples were collected at the end of ischemia (T-0), and 3 hours later (T-3) for assessment of testis malonaldehyde (MDA), reduced glutathione (GSH), and plasma total antioxidant power (TAP). RESULTS: MDA decreased significantly (p<0,001) in G-2 and G-3 in T-0 and T-3 timepoints. Additional decrease in MDA was seen in G-3 after 3 hours of reperfusion (T-3). GSH increased significantly in G-2 (p<0.001) and G-3 (p<0.05) at the end the ischemia (T-0). A significant increase in GSH was seen 3 hours after testis detorsion (T-3) in G-2 rats. TAP values remained unchanged. CONCLUSION: The data provides in vivo evidence of the antiperoxidative and antioxidative properties of TTN in torted rat testis.

OBJETIVO: Investigar o possível efeito protetor do bioflavonóide ternatina (TTN) quando administrado antes da indução da lesão de isquemia/reperfusão testicular em ratos. MÉTODOS: Trinta e seis ratos Wistar, aleatoriamente distribuídos em três grupos (n=12) divididos em dois subgrupos (n=6) cada foram tratados com solução salina (G-1), dimetilsulfóxido (DMSO) 3 por cento (G-2) ou TTN 12 mg/kg/dose (G-3), administrados i.p. 21, 12 e 1 hora antes da torção. Ratos anestesiados foram submetidos à isquemia (3 horas) induzida por torção (720º) do cordão espermático direito. Amostras (testículo ipsilateral e 3,0 ml de sangue arterial) foram coletadas ao final da isquemia (T-0), e 3 horas depois (T-3) para a avaliação das concentrações de malonaldeído (MDA), glutationa reduzida (GSH) no testículo e capacidade antioxidante total (TAP) no plasma. RESULTADOS: MDA diminuiu significativamente nos grupos G-2 e G-3 nos tempos T-0 e T-3. Houve diminuição adicional no G-3 após 3 horas. GSH aumentou significativamente nos grupos G-2 (p<0,001) e G-3 (p<0,05) no T-0 e T-3 no G-2. TAP permaneceu inalterada. CONCLUSÃO: Os achados fornecem evidências in vivo das propriedades antioxidantes e antiperoxidativas da TTN na T/D do testículo do rato.

Effect of short-term ornithine alpha-ketoglutarate pretreatment on intestinal ischemia-reperfusion in rats / Efeitos do pré-tratamento em curto prazo com ornitina alfa-cetoglutarato na isquemia-reperfusão intestinal em ratos

PURPOSE: To investigate the effects of preventive enteral administration of ornithine alpha-ketoglutarate (OKG) in an ischemia-reperfusion rat model. METHODS: Sixty rats were randomized into five groups (G1-G5, n = 12). Each group was divided into two subgroups (n = 6) and treated with calcium carbonate (CaCa) or OKG by gavage. Thirty minutes later, the animals were anesthetized with xylazine 15mg + ketamine 1mg ip and subjected to laparotomy. G1-G3 rats served as controls. Rats in groups G4 and G5 were subjected to ischemia for 30 minutes. Ischemia was achieved by clamping the small intestine and its mesentery, delimiting a segment of bowel 5 cm long and 5 cm apart from the ileocecal valve. In addition, G5 rats underwent reperfusion for 30 minutes. Blood samples were collected at the end of the laparotomy (G1), after 30 minutes (G2, G4) and 60 minutes (G3, G5) to determine concentrations of metabolites (pyruvate, lactate), creatine phosphokinase (CPK), thiobarbituric acid reactive substances (TBARS) and glutathione (GSH). RESULTS: There was a significant decrease in tissue pyruvate and lactate and plasma CPK levels in OKG-treated rats at the end of reperfusion period. GSH levels did not change significantly in ischemia and reperfusion groups. However, TBARS levels increased significantly (p<0.05) in tissue samples in OKG-treated rats subjected to ischemia for 30 minutes. CONCLUSION: Short-term pretreatment with OKG before induction of I/R decreases tissue damage, increases pyruvate utilization for energy production in the Krebs cycle and does not attenuate the oxidative stress in this animal model.

OBJETIVO: Investigar os efeitos da administração enteral preventiva de ornitina alfa-cetoglutarato (OKG) em modelo de isquemia-reperfusão no rato. MÉTODOS: Sessenta ratos foram randomizados em cinco grupos (G1-G5, n=12). Cada grupo foi redistribuído em dois subgrupos (n=6) e tratado com carbonato de cálcio (CaCa) ou OKG por gavagem. Trinta minutos mais tarde, os animais foram anestesiados com xilazina 1mg+cetamina 15mg i.p. e submetidos à laparotomia. Os ratos dos grupos G4-G5 foram submetidos à isquemia por 30 minutos. A isquemia foi obtida por pinçamento do intestino delgado, delimitando um segmento com 5 cm de comprimento e distando 5 cm da válvula ileocecal. O grupo G5 foi submetido à reperfusão por 30 minutos. Amostras de sangue foram coletadas no final da laparotomia (G1), após 30 minutos (G2, G4) e 60 minutos (G3, G5) para determinação das concentrações de metabolitos (piruvato, lactato), creatinofosfoquinase (CPK), substâncias reativas ao ácido tiobarbitúrico (TBARS) e glutationa (GSH). RESULTADOS: Observou-se redução significante (p<0,05) das concentrações de piruvato e lactato, teciduais e CPK plasmático em ratos tratados com OKG, no final do período de reperfusão. Não houve alteração significante nos níveis plasmáticos e teciduais de GSH. Entretanto os níveis de TBARS aumentaram significativamente (p<0,05) em amostras de tecido de ratos tratados com OKG submetido à isquemia por 30 minutos. CONCLUSÃO: o pré-tratamento em curto prazo com OKG antes da indução da I/R diminui a lesão tecidual, aumenta a utilização de piruvato para produção de energia no ciclo de Krebs, mas não atenua o estresse oxidativo neste modelo animal.

Preconditioning with L-alanyl-glutamine reduces hepatic ischemia-reperfusion injury in rats / Pré-condicionamento com L-alanil-glutamina reduz a lesão de isquemia-reperfusão no fígado de ratos

PURPOSE: To evaluate the effects of pre-conditioning with L-alanyl- glutamine (L-Ala-Gln) in rats subjected to total hepatic ischemia. METHODS: Thirty Wistar rats, average weight 300g, were randomly assigned to 3 groups (n=10): G-1 - Saline, G-2- L-Ala-Gln, G-3-control (Sham). G-1 and G-3 groups were treated with saline 2.0 ml or L-Ala-Gln (0.75mg/Kg) intraperitoneally (ip) respectively, 2 hours before laparotomy. Anesthetized rats were subjected to laparotomy and total hepatic ischemia (30 minutes) induced by by clamping of portal triad. Control group underwent peritoneal puncture, two hours before the sham operation (laparotomy only). At the end of ischemia (G1 and G2), the liver was reperfused for 60 minutes. Following reperfusion blood samples were collected for evaluation of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels. Liver (medium lobe) was removed for immunohistochemistry study with antibody for Caspase-3. RESULTS: It was found a significant decrease (p<0.05) of ALT levels (270.6 +40.8 vs 83.3 +5.5 - p <0.05), LDH (2079.0 +262.4 vs. 206.6 +16.2 - p <0.05) and Caspase-3 expression (6.72 +1.35 vs. 2.19 +1.14, p <0.05) in rats subjected to I / R, comparing the group treated with L-Ala -Gln with G-2. Also, the ALT level was significantly lower (P<0.05) in G-1 and G-2 groups than in G-3 (control group). CONCLUSION: L-Ala-Gln preconditioning in rats submitted to hepatic I/R significantly reduces ALT, LDH and Caspase-3 expression, suggesting hepatic protection.

OBJETIVO: Avaliar os efeitos do pré-condicionamento com L-alanil-glutamina (L-Ala-Gln) em ratos submetidos à isquemia hepática total. MÉTODOS: Trinta ratos Wistar, peso médio 300g foram divididos aleatoriamente em três grupos (n = 10): G-1 - Saline, G-2: L-Ala-Gln, G-3: controle. G-1 e G-3 grupos foram tratados com 2,0 ml de solução salina ou L-Ala-Gln (0,75 mg / kg) intraperitoneal (ip), respectivamente, duas horas antes da laparotomia. Ratos anestesiados foram submetidos à laparotomia e isquemia hepática total (30 minutos) induzida por pinçamento da tríade portal. O grupo controle foi submetido à punção peritoneal, duas horas antes da operação simulada (apenas laparotomia). No final da isquemia, o fígado foi reperfundido por 60 minutos. As amostras de sangue foram colhidas ao término da reperfusão para determinação das concentrações alanina aminotransferase (ALT) e desidrogenase láctica (LDH). O lobo médio do fígado foi removido para estudo imuno-histoquímico com anticorpo para caspase-3. RESULTADOS: Houve diminuição significante (p<0.05) dos valores de ALT (270,6+40,8 vs 83,3+5,5 - p<0,05), LDH (2079,0+262,4 vs 206,6+16,2 - p<0,05) e expressão da Caspase-3 (6,72+1,35 vs 2,19+1,14 -p<0,05) nos ratos submetidos à I/R, comparando o grupo tratado com L-Ala-Gln, ao grupo salina. Além disso, o nível de ALT foi significativamente menor (P <0,05) no G-1 e G-2 do que no grupo G-3 (grupo controle). CONCLUSÃO: O pré-condicionamento com L-Ala-Gln em ratos submetidos a I/R hepática reduz significativamente as concentrações de ALT e LDH e a expressão da caspase-3, sugerindo proteção hepática.

Preconditioning with L-alanyl-L-glutamine in a Mongolian Gerbil model of acute cerebral ischemia/reperfusion injury / Pré-condicionamento com L-alanil-L-glutamina em modelo de isquemia/reperfusão cerebral aguda em Gerbils da Mongólia

PURPOSE: To investigate the effect of L-alanyl-L-glutamine (L-Ala-Gln) preconditioning in an acute cerebral ischemia/reperfusion (I/R) model in gerbils. METHODS: Thirty-six Mongolian gerbils (Meriones unguiculatus), (60-100g), were randomized in 2 groups (n=18) and preconditioned with saline 2.0 ml (Group-S) or 0.75g/Kg of L-Ala-Gln, (Group-G) administered into the femoral vein 30 minutes prior to I/R. Each group was divided into three subgroups (n=6). Anesthetized animals (urethane, 1.5g/Kg, i.p.) were submitted to bilateral occlusion of common carotid arteries during 15 minutes. Samples (brain tissue and arterial blood) were collected at the end of ischemia (T0) and after 30 (T30) and 60 minutes (T60) for glucose, lactate, myeloperoxidase (MPO), thiobarbituric acid reactive substances (TBARS), glutathione (GSH) assays and histopathological evaluation. RESULTS: Glucose and lactate levels were not different in studied groups. However glycemia increased significantly in saline groups at the end of the reperfusion period. TBARS levels were significantly different, comparing treated (Group-G) and control group after 30 minutes of reperfusion (p<0.05) in cerebral tissue. Pretreatment with L-Ala-Gln promoted a significant increase in cerebral GSH contents in Group-G at T30 (p<0.001) time-point compared with Group-S. At T30 and T60, increased levels of GSH occurred in both time-points. There were no group differences regarding MPO levels. Pyknosis, presence of red neurons and intracellular edema were significantly smaller in Group-G. CONCLUSION: Preconditioning with L-Ala-Gln in gerbils submitted to cerebral ischemia/reperfusion reduces oxidative stress and degeneration of the nucleus (pyknosis) and cell death (red neurons) in the cerebral tissue.

OBJETIVO: Investigar o efeito do pré-condicionamento com L-alanil-L-glutamina (L-Ala-Gln) em gerbils submetidos à isquemia/reperfusão (I/R) cerebral aguda. MÉTODOS: Trinta e seis gerbils (Meriones unguiculatus) (60-100g) foram divididos em dois grupos (n=18) e pré-condicionados com 2,0 ml de soro fisiológico (Grupo-S) ou 0.75g/kg de L-Ala-Gln, (Grupo-G), administrados na veia femoral 30 minutos antes da I / R. Cada grupo foi dividido em três subgrupos (n=6).Animais anestesiados com uretano, 1.5g/kg, ip, foram submetidos à oclusão bilateral das artérias carótidas comuns, durante 15 minutos. Amostras (tecido cerebral e sangue arterial) foram coletadas no final da isquemia (T0) e após 30 (T30) e 60 minutos (T60) para a aferição das concentrações de glicose, lactato, mieloperoxidase (MPO), substâncias reagentes ao ácido tiobarbitúrico (TBARS), glutationa (GSH) e avaliação histopatológica. RESULTADOS: As concentrações de glicose e lactato não foram diferentes nos grupos estudados; a glicemia aumentou significativamente no Grupo-S ao final da reperfusão. Concentrações de TBARS no tecido cerebral foram significativamente diferentes, comparando os Grupos G e S, no T30 (p <0,05). O pré-tratamento com L-Ala-Gln promoveu um aumento significativo de GSH cerebral no Grupo-G comparado ao Grupo-S no T30 (p <0,001). Houve aumento das concentrações de GSH no T30 e T60 no Grupo-G. Não houve diferenças quanto as concentrações de MPO. Picnose, presença de neurônios vermelhos e edema intracelular foram significativamente menores no Grupo-G. CONCLUSÃO: O pré-condicionamento com L-Ala-Gln em gerbils submetidos à isquemia/reperfusão cerebral reduz o estresse oxidativo, a degeneração nuclear (picnose) e morte celular (neurônios vermelhos) no tecido cerebral.

L-Alanyl-Glutamine dipeptide pretreatment attenuates ischemia-reperfusion injury in rat testis / Pré-tratamento com o dipeptídeo L-Alanil-glutamina atenua a lesão por isquemia e reperfusão no testículo do rato

PURPOSE: To investigate the effect of alanyl-glutamine dipeptide (L-Ala-Gln) pre-treatment on ischemia-reperfusion (I/R) injury after unilateral testicular torsion-detorsion in a comparative controlled experiment. METHODS: Forty-eight rats (150-200 g) randomly distributed into 4 groups (n=12), and distributed in 2 subgroups (n=6) each, were treated with saline 2.0 ml (G-1, G-3) or L-Ala-Gln 20 percent, 0.75g/kg dissolved in saline (total volume 2.0 ml) administered in the left saphenous vein 30 minutes before ischemia. Anesthetized rats were subjected to I/R induced by torsion (720°) of the right spermatic cord lasting 1h (G-1, G-2) or 3 hours (G-3, G4). Anesthesia was again applied at the end of ischemia time (T-0) for testis detorsion and 6 hours later (T-6) for orchiectomy. All operations were performed on the right testes through transverse scrotal incisions. Right orchiectomy was carried out at the end of ischemia (T-0), and 6 hours later (T-6) to evaluate the concentrations of malondialdehyde (MDA) and reduced glutathione (GSH) in the testis. RESULTS: Pretreatment with L-Ala-Gln reduced MDA contents in rat testis at the end of ischemia lasting 3 hours. There was significant increase of GSH levels in T-6 time-point after 1 hour of ischemia. GSH levels also increased in T-0 and T-6 time-points in rats subjected to ischemia for 3 hours. CONCLUSION: L-Ala-Gln administered before torsion/detorsion of the spermatic cord decreases lipid peroxidation during ischemia and protects the testis from oxidative stress by upregulating GSH levels during reperfusion.

OBJETIVO: Investigar o efeito do pré-tratamento com o dipeptídeo L-alanil-glutamina (L-Ala-Gln) sobre a lesão de isquemia e reperfusão (I/R), induzida por torção/destorção do testículo em um experimento controlado e comparativo. MÉTODOS: Quarenta e oito ratos (150-200 g) divididos em quatro grupos (n=12) e distribuídos em dois subgrupos (n = 6) cada, foram tratados com 2,0 ml de solução salina (G-1, G-3 ) ou L-Ala-Gln 20 por cento, 0,75g/kg dissolvida em solução salina (volume total de 2,0 ml), administrada na veia safena 30 minutos antes da isquemia. Ratos anestesiados foram submetidos à torção (720°) do cordão espermático direito durante 1h (G-1, G-2) ou 3 horas (G-3, G4) para indução da I/R. A anestesia foi reaplicada no final do tempo de isquemia (T-0) para destorção do testículo e 6 horas depois (T-6) para orquiectomia. Todas as operações foram realizadas nos testículos direitos através de incisões escrotais. Orquiectomia direita foi realizada no final de isquemia (T-0), e seis horas depois (T-6) para avaliar as concentrações de malondialdeído (MDA) e glutationa reduzida (GSH) no testículo. RESULTADOS: O pré-tratamento com L-Ala-Gln reduziu os níveis de MDA no testículo de ratos no final da isquemia (3 horas). Entretanto os níveis de GSH aumentaram significativamente no T-6 após 1 hora de isquemia e também no T-0 e T-6 em ratos submetidos à isquemia por 3 horas. CONCLUSÃO: L-Ala-Gln administrada antes da torção/destorção do cordão espermático diminui a peroxidação lipídica na isquemia e protege o testículo contra o estresse oxidativo, promovendo aumento dos níveis de GSH durante a reperfusão.

Preconditioning with oil mixes of high ratio Omega-9: Omega-6 and a low ratio Omega-6:Omega-3 in rats subjected to brain ischemia/reperfusion / Pré-condicionamento com misturas de óleos com Ômega-9: Ômega-6 (alta relação) e Ômega-6:Ômega-3 (baixa relação) em ratos submetidos à isquemia/reperfusão cerebral

PURPOSE: This study aimed to assess the effects of preconditioning with mixtures of oils containing high/low ratio of ω-6/ω-3 and ω-9/ω-6, respectively, in an experimental model of cerebral ischemia-reperfusion (I/R). METHODS: Forty-two Wistar rats were randomly distributed into two groups: control (n=24) and test (n=18). Control group was subdivided in 4 subgroups (n=6): G1: Sham-Water; G2: I/R-Water; G3: Sham-Isolipidic and G4: I/R-Isolipid. The animals received water or a isolipid mixture containing ω-3 oils (8:1 ratio) and ω-9/ω-6 (0.4:1 ratio) by gavage for seven days. Test group included 3 subgroups (n=6) G5: I/R-Mix1, G: 6 I/R-Mix2 and G7: I/R-Mix3. Test group animals received oily mixtures of ω-3 (1.4:1 ratio) and ω-6 (3.4:1 ratio), differing only in source of ω-3: G5 (alpha-linolenic acid); G6 (alpha-linolenic, docosahexaenoic and eicosapentaenoic acids), and G7 (alpha-linolenic and docosahexaenoic acids). On day 7 I/R rats underwent cerebral ischemia with bilateral occlusion of common carotid arteries for 1 hour followed by reperfusion for 3 hours. G1 and G3 animals underwent sham operation. Concluded the experiment, animals were decapitated and their brains sliced for red neurons (RN) count in CA3 area of the hippocampus. Variables were compared using ANOVA-Tukey test. RESULTS: The use of different mix preparations promoted a decrease in red cell count in all three groups (G5/G6/G7), compared with G2/G4, confirming the protective effect of different oil blends, regardless of ω-3 source. CONCLUSION: Pre-conditioning with mixtures of oils containing high ratio ω-6/ω-3 and low ω-9/ω-6 relationship protects brain neurons against I/R injury in an experimental model.

OBJETIVO: Avaliar os efeitos do pré-condicionamento com misturas de óleos contendo relação alta/baixa de ω-6/ω-3 e ω-9/ω-6, respectivamente, em um modelo experimental de isquemia/reperfusão (I/R) cerebral. MÉTODOS: Quarenta e dois ratos foram distribuídos aleatoriamente em dois grupos: controle (n=24) e teste (n=18). Grupo controle foi subdividido em quatro subgrupos (n=6): G1: Sham-Água; G2: I/R-Água; G3: Sham-Isolipídico e G4: I/R-Isolipídico. Os animais receberam água ou uma mistura isolipidica contendo ω-6/ω-3 óleos (8:1) e ω-9/ω-6 (0,4:1) por gavagem, durante sete dias. O grupo teste incluiu três subgrupos (n=6) G5: I/R-Mix1, G: 6 I/R-Mix2 e G7: I/R-Mix3. Animais do grupo teste receberam de misturas de óleos ω-6/ω-3 (1,4:1) e ω-9/ω-6 (3,4:1), diferindo apenas na fonte de -3: G5:alpha-linolênico; G6: ácidos alpha-linolênico, eicosapentaenóico e docosahexaenóico e G7:ácidos alpha-linolênico e docosahexaenóico. No 7º dia os grupos I/R foram submetidos à isquemia cerebral (1h) por oclusão bilateral das artérias carótidas comuns seguida de reperfusão (3h). Ratos G1 e G3 foram submetidos à operação simulada. Concluído o experimento, os animais foram decapitados e seus cérebros fatiados para contagem dos neurônios vermelhos na área CA3 do hipocampo. As variáveis foram comparadas pelo teste de ANOVA-Tukey. RESULTADOS: A utilização de diferentes misturas de óleos promoveu uma diminuição na contagem de células vermelhas nos grupos G5/G6/G7, em comparação com G2/G4, confirmando o efeito protetor das misturas de óleos, independentemente da origem de ω-3. CONCLUSÃO: O pré-condicionamento com misturas de óleos contendo alta proporção de ω-6/ω-3 e baixa proporção de ω-9/ω-6 protege os neurônios cerebrais da lesão de I/R em um modelo experimental.