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.

New metallophamaceutic reduced renal injury induced by non-steroidal anti-inflammatory

Abstract Purpose To evaluate the effect of Rut-bpy (Cis-[Ru(bpy)2(SO3)(NO)]PF 6), a novel nitric oxide donor, able to modulate the histological changes caused by the NASID (meloxicam). Methods Wistar rats were assigned into three groups (n=6 rats/group): Sham group (saline solution), NSAID group (meloxicam - 15 mg/kg) and Rut-bpy group (100 mg/kg of Rut-bpy associated with 15mg/kg of meloxicam). At the end of experiments, kidneys were removed for histological study, fractal dimension and lacunarity in all animals. Results At the histological examination, all animals (six animals - 100 %) in the NSAID group had membrane thickening and other changes (necrosis, acute tubular congestion and vascular congestion); on the other hand, only one animal (16.6 %) of the Rut-bpy group had congestion. The fractal dimension and lacunarity were greater in the control and Rut-bpy group than in NSAIDs group (p<0.05). Conclusion Rut-bpy may prevent renal histological changes in rats caused by meloxicam.

Efeitos do nitrosil rutênio na lesão cerebral induzida porisquemia e reperfusão em ratos

Efeitos do nitrosil rutênio na lesão cerebral induzida por isquemia e reperfusão em ratos. MARCIO WILKER SOARES CAMPELO. Pós-Graduação Stricto Sensu do Departamento de Cirurgia, Faculdade de Medicina, Universidade Federal do Ceará (Grau de Mestre em Cirurgia). Novembro, 2009. Orientador: Prof. Dr. Paulo Roberto Leitão de Vasconcelos. Introdução e objetivo: Doadores de NO podem diminuir a lesão neuronal durante a isquemia e reperfusão cerebral (I/R) por aumento do fluxo sanguíneo cerebral. O objetivo deste estudo é avaliar se um novo complexo de nitrosilo complexo de rutênio (Rut-bpy) capaz liberar NO direto na musculatura lisa vascular apresenta algum efeito durante I/R. Método: Foram utilizados 96 ratos machos, da linhagem Wistar, com peso médio de 290.27 g, distribuídos em 2 fases com 8 grupos cada: Fase de isquemia 4 grupos sham (SF, Rut-bpy,L-NAME e L-NAME+Rut-bpy) e 4 grupos isquemia (SF, Rut-bpy,L-NAME e L-NAME+Rut-bpy); da mesma forma foi dividido a fase de reperfusão ( 4 grupos sham e 4 grupos isquemia/reperfusão) com as mesmas drogas teste da fase de isquemia. Foi utilizado um modelo de isquemia cerebral global incompleta, com oclusão da artéria carótida comum bilateral e administração do SF, Rut-bpy e L-NAME via intraperitoneal. No final do experimento os animais foram decapitados e o cérebro fatiado para ser avaliado à área de lesão por histoquímica. Durante todo o experimento a PAM dos animais foi monitorizada...

Effects of nitrosyl ruthenium in injury brain induced by ischemia and reperfusion of rats.MARCIO WILKER SOARES CAMPELO. Stricto SensuPost-graduation. Department of Surgery, Medicine School, Federal University of Ceará (Degree of Master of Surgery). November, 2009. Advisor: Prof. Dr. Paulo Roberto Leitão de Vasconcelos. Background and purpose - Nitric oxide (NO) donors are known to reduce neuronal damage during brain ischemia and reperfusion by increasing the blood flow. Rut-bpy is a novel nitrosyl-ruthenium complex releasing NO directly into the vascular smooth musculature. The objective of the study was to evaluate the effect of Rut-bpy on a rat model of brain ischemia and reperfusion. Methods - Ninety-six male Wistar rats weighing approximately 290g were randomly assigned to 16 groups. Four groups and their respective sham groups were submitted to ischemia (Stage 1), while four groups and their respective sham groups were submitted to ischemia + reperfusion (Stage 2). At each stage of the experiment the groups were treated pairwise with saline solution (SS), Rut-bpy, L-NAME and L-NAME+Rut-bpy, respectively. The study was based on an incomplete global brain ischemia model with occlusion of the common bilateral carotid arteries and intraperitoneal administration of the study drugs. Following the experiment the animals were decapitated and the brain was sectioned for histochemical evaluation of the area of damage. The mean arterial blood pressure (MABP) was monitored throughout the experiment...

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. .

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. .

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. .

An optimized animal model for partial and total skin thickness burns studies / Um modelo animal aperfeiçoado para estudo de queimaduras superficiais e profundas da pele

PURPOSE: Development of an improved animal model for studying skin burns in rats. METHODS: Twenty-four male Wistar rats were randomly assigned to four groups (n=6): G1-Control, G2- T100°C, G3-T150°C and G4-T200°C. Two 10 x 10 mm squares were outlined with a sterile surgical marker on each side and along the vertebral column using a prepared template positioned between the anterior and posterior limbs. G2-G4 rats were subjected to 100°C, 150°C and 200ºC thermal burns, respectively. G1 rats served as controls. Burns were inflicted by applying a copper plate connected to an electronic temperature controlling device to the dorsal skin of anesthetized rats. Four burns were produced on each animal (total area: 4 cm²/animal) leaving about 1 cm of undamaged skin between burn areas. Analgesia was administered during 24 h after burn injury by adding 30 mg codeine phosphate hemihydrate to 500 ml tap water. RESULTS: The application of 100°C and 150ºC resulted in partial thickness skin burns with central reepithelialization of the burned area only at 100°C. In G4 group the whole thickness of the skin was injured without central reepithelialization. However, there was marginal reepithelialization in all groups. CONCLUSION: The model studied is inexpensive and easily reproducible, enabling the achievement of controlled burns with partial or total impairment of the skin in experimental animals.

OBJETIVO: Desenvolvimento de um modelo animal aperfeiçoado para estudo de queimaduras cutâneas em ratos. MÉTODOS: Vinte e quatro ratos Wistar, machos, foram distribuídos aleatoriamente em quatro grupos (n=6): G1-Controle, G2-T100°C, G3-T150°C e G4-T200°C. Dois quadrados medindo 10x10 mm foram delineados com um marcador cirúrgico estéril em cada lado e ao longo da coluna vertebral e posicionados entre os membros anteriores e posteriores, utilizando um molde previamente preparado. Os ratos dos grupos G2-G4 foram submetidos a queimaduras térmicas de 100°C, 150°C e 200°C, respectivamente. O grupo G1 foi utilizado como controle. As queimaduras foram infligidas pela aplicação de uma placa de cobre, ligada a um dispositivo de controle eletrônico de temperatura, na pele dorsal de ratos anestesiados. Quatro queimaduras foram produzidas em cada animal (área total: 4 cm2/animal), deixando cerca de 1 cm de pele intacta entre as áreas queimadas. Analgesia foi obtida durante 24 horas após a queimadura por adição de 30mg de fosfato hemi-hidratado de codeína a 500 ml de água potável. RESULTADOS: A aplicação 100°C e 150°C resultou na produção de queimaduras profundas comprometendo parte da espessura da pele, com reepitelização central da área queimada, somente a 100°C. No grupo G4 houve lesão de toda a espessura da pele sem reepitelização central. Entretanto, observou-se reepitelização marginal em todos os grupos estudados. CONCLUSÃO: O modelo estudado é de baixo custo e facilmente reproduzível, propiciando a obtenção controlada de queimaduras com comprometimento parcial ou total da pele, em animais experimentais.

Effects of Rut-bpy (Cis-[Ru(bpy)2(SO3)(NO)]PF 6), a novel nitric oxide donor, in L-NAME-induced hypertension in rats / Efeitos do Rut-bpy (Cis-[Ru(bpy)2(SO3)(NO)]PF 6), um novo doador de óxido nítrico, na hipertensão induzida com L-NAME em ratos

PURPOSE: To evaluate the effect of Rut-bpy (Cis-[Ru(bpy)2(SO3)(NO)]PF 6), a novel nitric oxide donor in Nω-nitro-L-arginine methyl ester (L-NAME)-induced hypertensive rats. METHODS: Twenty-four male Wistar rats were randomly assigned to four groups (n=6), named according to the treatment applied (G1-Saline, G2-Rut-bpy, G3-L-NAME and G4-L-NAME+Rut-bpy). L-NAME (30 mg/Kg) was injected intraperitoneally 30 minutes before the administration of Rut-bpy (100 mg/Kg). Mean abdominal aorta arterial blood pressure (MAP) was continuously monitored. RESULTS: Mean arterial blood pressure (MAP) in G3 rats rose progressively, reaching 147±16 mmHg compared with 100±19 mm Hg in G1 rats (p<0.05). In G4 rats, treated with L-NAME+Rut-bpy, MAP reached 149+11 mm Hg while in G2 rats, treated with Rut-bpy, MAP values were 106±11 mm Hg. In G1 rats these values decreased progressively reaching 87+14 mm Hg after 30 minutes. An important finding was the maintenance of the MAP throughout the experiment in G2 rats. CONCLUSION: Rut-bpy does not decrease the MAP in L-Name induced hypertensive rats. However, when it is used in anesthetized hypotensive rats a stable blood pressure is obtained.

OBJETIVO: Avaliar o efeitos do Rut-bpy (Cis-[Ru (bpy)2(SO3)(NO)] PF6), um novo doador de óxido nítrico, em ratos hipertensos induzidos pelo éster metílico de N-nitro-L-arginina (L-NAME). MÉTODOS: Vinte e quatro ratos Wistar machos foram distribuídos aleatoriamente em quatro grupos (n = 6), nomeados de acordo com o tratamento aplicado (G1-Salina, G2-Rut-bpy, G3-L-NAME e G4-L-NAME+Rut -bpy). L-NAME (30 mg / Kg) foi injetado por via intraperitoneal 30 minutos antes da administração de Rut-bpy (100 mg / kg). A pressão arterial média (PAM) da aorta abdominal foi monitorada continuamente. RESULTADOS: A pressão arterial média (PAM) em ratos do grupo G3 subiu progressivamente, chegando a 147 ±16 mm Hg, em comparação com 100 ±19 mm Hg em ratos do G1 (p <0,05). Em ratos G4, tratados com L-NAME + Rut-bpy, a PAM atingiu 149±11 milímetros de Hg, enquanto no G2 (ratos tratados com Rut bpy) os valores da PAM foram 106 ±11 mm Hg. No G1 esses valores decresceram progressivamente, atingindo 87±14 mm Hg após 30 minutos. Um achado importante foi a manutenção da PAM durante todo o experimento em ratos do grupo G2. CONCLUSÃO: O uso de Rut bpy não diminui a PAM em ratos hipertensos por L-NAME. No entanto, quando ele é usado em ratos anestesiados, hipotensos, uma pressão arterial estável é obtida.