RESUMO
The aim of this study was to evaluate the effects of single oral doses of D-005 (a lipid extract obtained from the fruit oil of Acrocomia crispa) on LPS-induced acute lung injury (ALI) in mice. D-005 batch composition was: lauric (35.8%), oleic (28.4%), myristic (14.2%), palmitic (8.9%), stearic (3.3%), capric (1.9%), caprylic (1.2%), and palmitoleic (0.05%) acids, for a total content of fatty acids of 93.7%. D-005 (200 mg/kg) significantly reduced lung edema (LE) (≈ 28% inhibition) and Lung Weight/Body Weight ratio (LW/BW) (75.8% inhibition). D-005 (25, 50, 100 and 200 mg/kg) produced a significant reduction of Histological score (59.9, 56.1, 53.5 and 73.3% inhibition, respectively). Dexamethasone, as the reference drug, was effective in this experimental model. In conclusion, pretreatment with single oral doses of D-005 significantly prevented the LPS-induced ALI in mice.
El objetivo de este estudio fue evaluar los efectos de dosis orales uÌnicas de D-005 (extracto lipiÌdico obtenido del aceite de frutos de Acrocomia crispa) sobre el danÌo pulmonar agudo (DPA) inducido por LPS en ratones. La composicioÌn del lote de D-005 fue: aÌcido laÌurico (35.8%), oleico (28.4%), miriÌstico (14.2%), palmiÌtico (8.9%), esteaÌrico (3.3%), caÌprico (1.9%), capriÌlico (1.2%) y palmitoleico (0.05%), con un contenido total de aÌcidos grasos de 93.7%. D-005 (200 mg/kg) redujo significativamente el edema pulmonar (EP) (≈ 28% de inhibicioÌn) y la relacioÌn peso pulmoÌn/peso corporal (PP/PC) (75.8% de inhibicioÌn). D-005 (25, 50, 100 y 200 mg/kg) produjo una reduccioÌn significativa de la puntuacioÌn histoloÌgica (59.9, 56.1, 53.5 y 73.3% de inhibicioÌn, respectivamente). La dexametasona, faÌrmaco de referencia, fue efectiva en este modelo experimental. En conclusioÌn, el pretratamiento con dosis orales uÌnicas de D-005 previno significativamente el DPA inducido por LPS en ratones.
Assuntos
Animais , Camundongos , Extratos Vegetais/administração & dosagem , Arecaceae , Lesão Pulmonar Aguda/prevenção & controle , Extratos Vegetais/química , Lipopolissacarídeos/efeitos adversos , Administração Oral , Cromatografia Gasosa , Lesão Pulmonar Aguda/induzido quimicamente , Ácidos Graxos/análise , Frutas , Pulmão/efeitos dos fármacosRESUMO
OBJECTIVE: The aim of this study was to evaluate the antinociceptive activity of the methanol extract of Tabebuia hypoleuca stems (THME). MATERIALS AND METHODS: The animals were divided into 5 groups of 8 mice for each test (negative controls, positive controls, and 3 groups treated with THME at doses of 150, 300, and 500 mg/kg, p.o.). The antinociceptive effect of THME was evaluated using the writhing, formalin, tail flick, and hot plate models in mice. RESULTS: In the writhing test, THME (150, 300, and 500 mg/kg) produced significantly (p < 0.001) fewer writhes induced by acetic acid than in the control group. In the formalin test, the licking time for THME at doses of 300 and 500 mg/kg was signiï¬cantly shorter (p < 0.001) compared to the control group in the first phase of the formalin test, whereas in the second phase only the dose of 500 mg/kg showed an antinociceptive effect. In addition, THME at doses of 300 and 500 mg/kg significantly increased the latency time in the tail flick test (p < 0.05 and p < 0.001, respectively) and in the hot plate test (p < 0.01 and p < 0.001, respectively) compared to the control group. CONCLUSIONS: These results show that THME had antinociceptive activity using several models of nociception, and they suggest that the effect is mediated by the participation of both peripheral and central antinociceptive mechanisms.
Assuntos
Analgésicos/farmacologia , Extratos Vegetais/farmacologia , Tabebuia/efeitos dos fármacos , Ácido Acético , Análise de Variância , Animais , Cuba , Feminino , Masculino , Metanol , Camundongos , Camundongos Endogâmicos BALB C , Dor/tratamento farmacológico , Medição da Dor , Ratos , Ratos Sprague-Dawley , Tabebuia/toxicidade , CaudaRESUMO
One important goal of cancer immunotherapy is to prevent and treat tumor metastasis. We have previously reported the significant antitumor effect induced by the immunization with our human papillomavirus therapeutic protein-based vaccine (LALF32-51-E7) without adjuvant and admixed with clinically relevant adjuvants in the subcutaneous TC-1 tumor challenge model. In the present study, we evaluated the efficacy of the above mentioned vaccine formulations in controlling the hematogenous spread of TC-1 tumor cells using a more tumourigenic clone named TC-1* and other intravenous injection site less stressful than the tail vein. We generated a lung metastasis model by injecting TC-1* cells into the retro-orbital venous sinus and this is the first study describing it. Also, this is the first study that demonstrates the efficacy of the immunization with LALF32-51-E7 without adjuvant and admixed with VSSP or Al(OH)3 in controlling metastatic tumors increasing the survival of the mice. Our TC-1 lung metastasis model can be used to test the efficacy of other immunotherapeutic strategies based on E6/E7 antigens.
Assuntos
Imunoterapia , Neoplasias Pulmonares/secundário , Neoplasias Pulmonares/terapia , Proteínas E7 de Papillomavirus/imunologia , Vacinas contra Papillomavirus/uso terapêutico , Neoplasias do Colo do Útero/terapia , Animais , Feminino , Vetores Genéticos , Humanos , Neoplasias Pulmonares/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Proteolipídeos , Linfócitos T Citotóxicos , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/imunologia , Neoplasias do Colo do Útero/patologiaRESUMO
The development of therapeutic vaccines against chronic hepatitis B requires the capacity of the formulation to subvert a tolerated immune response as well as the evaluation of histopathological damage resulting from the treatment. In the present study, the dynamicity of induced immune response to hepatitis B surface antigen (HBsAg) was evaluated in transgenic mice that constitutively express the HBsAg gene (HBsAg-tg mice). After immunization with a vaccine candidate containing both surface (HBsAg) and core (HBcAg) antigens of hepatitis B virus (HBV), the effect of vaccination on clearance of circulating HBsAg and the potential histological alterations were examined. Transgenic (tg) and non-transgenic (Ntg) mice were immunized by intranasal (IN) and subcutaneous (SC) routes simultaneously. A control group received phosphate-buffered saline (PBS) by IN route and aluminum by SC route. Positive responses, at both humoral and cellular levels, were obtained after five immunizations in HBsAg-tg mice. Such responses were delayed and of lower intensity in tg mice, compared to vaccinated Ntg mice. Serum IgG response was characterized by a similar IgG subclass pattern. Even when HBsAg-specific CD8+ T cell responses were clearly detectable by gamma-interferon ELISPOT assay, histopathological alterations were not detected in any organ, including the liver and kidneys. Our study demonstrated, that it is possible to subvert the immune tolerance against HBsAg in tg mice, opening a window for new studies to optimize the schedule, dose, and formulation to improve the immune response to the therapeutic vaccine candidate. These results can be considered a safety proof to support clinical developments for the formulation under study. HOW TO CITE THIS ARTICLE: Freyre FM, Blanco A, Trujillo H, Hernández D, García D, Alba JS, Lopez M, Merino N, Lobaina Y, Aguilar JC. Dynamic of Immune Response induced in Hepatitis B Surface Antigen-transgenic Mice Immunized with a Novel Therapeutic Formulation. Euroasian J Hepato-Gastroenterol 2016;6(1):25-30.
RESUMO
The present study examines the possible effect of the glucosylxanthone mangiferin (MG) on pain-related behaviors in a tonic acute pain model (formalin test at 5%) and in a chronic constriction injury (CCI) model to clarify the underlying transient and long-term mechanisms. Acute administration of MG (10-100mg/kg, i.p.) reduced licking/biting exclusivity in the tonic phase of formalin test in a naloxone and yohimbine-sensitive manner. This effect was enhanced by a nonselective nitric oxide synthase (NOS) inhibitor (NG-monomethyl-L-arginine) and by a non-competitive N-methyl-D-aspartate (NMDA) antagonist (ketamine), but it was reversed by the NOS substrate (L-arginine). Pre-treatment with intrathecal yohimbine prevented the anti-hypernociceptive effect of systemic MG. Pre-treatment during 4 days before surgical and 3 days after CCI with MG (50mg/kg, i.p.) reduced mechanical hypernociception and decreased the signs of Wallerian degeneration (WD) of the sciatic nerve. MG improved the PC-12 cellular viability exposure to glutamate-mediated neuronal death, also involved in neuropathic pain. The findings of this study suggest that MG shows ability to decrease tonic pain in the formalin test. A transient activity of this xanthone on nociceptive pathways mediated by α2 adrenergic receptors in cooperation with the opioid system could be involved, at least in part, in this effect. Its neuroprotective effect by preventing WD in mononeuropathic rats could be implicated in the mechano-antihypernociceptive long term mechanisms.
Assuntos
Analgésicos/uso terapêutico , Modelos Animais de Doenças , Neuralgia/tratamento farmacológico , Xantonas/uso terapêutico , Animais , Comportamento Animal/efeitos dos fármacos , Masculino , Células PC12 , Ratos , Ratos Sprague-Dawley , Xantonas/farmacologiaRESUMO
We previously showed that JM-20, a novel 1,5-benzodiazepine fused to a dihydropyridine moiety, possessed an anxiolytic profile similar to diazepam and strong neuroprotective activity in different cell models relevant to cerebral ischemia. Here, we investigated whether JM-20 protects against ischemic neuronal damage in vitro and in vivo. The effects of JM-20 were evaluated on hippocampal slices subjected to oxygen and glucose deprivation (OGD). For in vivo studies, Wistar rats were subjected 90 min of middle cerebral artery occlusion (MCAo) and oral administration of JM-20 at 2, 4 and 8 mg/kg 1 h following reperfusion. Twenty-four hours after cerebral blood flow restoration, neurological deficits were scored, and the infarct volume, histopathological changes in cortex, number of hippocampal and striatal neurons, and glutamate/aspartate concentrations in the cerebrospinal fluid were measured. Susceptibility to brain mitochondrial swelling, membrane potential dissipation, H2O2 generation, cytochrome c release, Ca2+ accumulation, and morphological changes in the organelles were assessed 24 h post-ischemia. In vitro, JM-20 (1 and 10 µM) administered during reperfusion significantly reduced cell death in hippocampal slices subjected to OGD. In vivo, JM-20 treatment (4 and 8 mg/kg) significantly decreased neurological deficit scores, edema formation, total infarct volumes and histological alterations in different brain regions. JM-20 treatment also protected brain mitochondria from ischemic damage, most likely by preventing Ca2+ accumulation in organelles. Moreover, an 8-mg/kg JM-20 dose reduced glutamate and aspartate concentrations in cerebrospinal fluid and the deleterious effects of MCAo even when delivered 8 h after blood flow restoration. These results suggest that in rats, JM-20 is a robust neuroprotective agent against ischemia/reperfusion injury with a wide therapeutic window. Our findings support the further examination of potential clinical JM-20 use to treat acute ischemic stroke.
Assuntos
Benzodiazepinas/farmacologia , Isquemia Encefálica/tratamento farmacológico , Aminoácidos Excitatórios/metabolismo , Mitocôndrias/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Niacina/análogos & derivados , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Encéfalo/fisiopatologia , Isquemia Encefálica/patologia , Isquemia Encefálica/fisiopatologia , Morte Celular/efeitos dos fármacos , Morte Celular/fisiologia , Modelos Animais de Doenças , Glucose/deficiência , Infarto da Artéria Cerebral Média/tratamento farmacológico , Infarto da Artéria Cerebral Média/fisiopatologia , Masculino , Mitocôndrias/patologia , Mitocôndrias/fisiologia , Niacina/farmacologia , Distribuição Aleatória , Ratos Wistar , Técnicas de Cultura de TecidosRESUMO
OBJECTIVES: The aim was to study the effects of Mangifera indica extract and its major component mangiferin on lung inflammation response and Th2 cytokine production using a murine experimental model of allergic asthma. METHODS: BALB/c mice were intraperitoneally sensitized with 10 µg of ovoalbumin (OVA) adsorbed on aluminium hydroxide on days 0, 7 and 14. Seven days after the last injection, the mice were challenged with 2% aerosolized OVA inhalation for 30 min beginning on day 21 and continuing until day 24. To evaluate the protective effect, mice were orally treated with M. indica extract (50, 100 or 250 mg/kg) or mangiferin (50 mg/kg) from days 0 to 24. Anti-OVA immunoglobulin E, interleukin (IL)-4 and IL-5 were determined by ELISA and lungs were analysed by histology. KEY FINDINGS: M. indica extract and mangiferin produced a marked reduction of airway inflammation around vessels and bronchi, inhibition of IL-4 and IL-5 cytokines in bronchoalveolar lavage fluid and lymphocyte culture supernatant, IgE levels and lymphocyte proliferation. CONCLUSION: This is the first pre-clinical report of the anti-inflammatory properties of M. indica extract and mangiferin in experimental asthma and it could be an important part of pre-clinical requirement necessary for its use to complement the treatment of this complex disease.
Assuntos
Anti-Inflamatórios/uso terapêutico , Asma/tratamento farmacológico , Inflamação/tratamento farmacológico , Mangifera/química , Fitoterapia , Extratos Vegetais/uso terapêutico , Xantonas/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Asma/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Imunoglobulina E/metabolismo , Inflamação/imunologia , Interleucina-4/metabolismo , Interleucina-5/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Linfócitos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina , Casca de Planta , Extratos Vegetais/farmacologia , Caules de Planta , Células Th2/metabolismo , Xantonas/isolamento & purificação , Xantonas/farmacologiaRESUMO
Mango (Mangifera indica L.) stem bark aqueous extract (MSBE) is a natural product with antioxidant, anti-inflammatory, analgesic, and immunomodulatory effects. Its formulations (e.g., tablets, capsules, syrup, vaginal oval, and suppositories) are known by the brand name of Vimang. In view of the ethnomedical, preclinical, and clinical uses of this extract and the necessity to assess its possible toxicological effect on man, a toxicological analysis of a standard extract is reported in this paper. Acute toxicity was evaluated in mice and rats by oral, dermal, and intraperitoneal (i.p.) administration. The extract, by oral or dermal administration, showed no lethality at the limit doses of 2,000 mg/kg body weight and no adverse effects were found. Deaths occurred with the i.p. administration at 200, but not 20 mg/kg in mice. MSBE was also studied on irritant tests in rabbits, and the results showed that it was nonirritating on skin, ocular, or rectal mucosa. The extract had minimal irritancy following vaginal application.
Assuntos
Antioxidantes/toxicidade , Mangifera/química , Extratos Vegetais/toxicidade , Administração Cutânea , Administração Oral , Animais , Antioxidantes/administração & dosagem , Antioxidantes/isolamento & purificação , Relação Dose-Resposta a Droga , Feminino , Injeções Intraperitoneais , Masculino , Medicina Tradicional , Camundongos , Casca de Planta , Extratos Vegetais/administração & dosagem , Coelhos , Ratos , Ratos Sprague-Dawley , Testes de Toxicidade AgudaRESUMO
Central sensitization theory has been defined as pivotal for understanding the excitability changes in central neurons following peripheral inflammation or neuropathic injury. Considerable evidence has demonstrated that activation of the N-methyl-D-aspartate (NMDA) subtype of glutamate receptors and subsequent nitric oxide (NO) production are the key in these changes. Consequently, neuromodulator drugs have been developed during the last decades. The electroacupuncture (EA) that acts as biochemical modulator in the spinal horn cord would prevent these changes. The aim of this study was to determine the thermal anti-hyperalgesic effect of EA (10 Hz, 3 mA) and its combination with L-NAME as nitric oxide synthase (NOS) inhibitor in carrageenan-induced hyperalgesia in rats. Also, it investigated the changes in the plasmatic concentrations of NO metabolites. Moreover, the EA combination with sub-effective dose of ketamine as a NMDA antagonist was tested. The EA pre-treatment conducted in unsedated, unrestrained and conscious animals showed a thermal anti-hyperalgesic effect in correspondence with plasmatic increase of NO metabolites. The L-NAME (30 mg/kg) pre-administration decreased significantly the plasmatic concentrations of NO(2)(-)/NO(3)(-) and suppressed the anti-hyperalgesic effect of EA. The combination of EA with ketamine enhanced the anti-hyperalgesic effect. These data constitute the first report that suggested the participation, at least in part, of the L-arginine-NOS-NO-GMPc pathway activation in anti-hyperalgesic effect of EA in carrageenan-induced inflammation model.
Assuntos
Eletroacupuntura , Inflamação/metabolismo , Inflamação/terapia , Óxido Nítrico/metabolismo , Animais , Carragenina , Inibidores Enzimáticos/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Temperatura Alta , Hiperalgesia/induzido quimicamente , Hiperalgesia/metabolismo , Hiperalgesia/terapia , Inflamação/induzido quimicamente , Ketamina/farmacologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Nitratos/sangue , Óxido Nítrico Sintase/antagonistas & inibidores , Dióxido de Nitrogênio/sangue , Medição da Dor , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVES: The ischaemia-reperfusion process is largely mediated by reactive oxygen species. Taking into account that a transient and controlled administration of ozone is able to upregulate cellular antioxidant enzymes, a morphological, biochemical and functional renal study was performed in rats undergoing warm renal ischaemia. METHODS: Rats were divided into four groups. All except the negative controls underwent 60 min' bilateral renal ischaemia followed by 10 days' reperfusion. The positive control group received no further treatment. The ozone group received an ozone/oxygen mixture (ozone dose 0.5 mg/kg) immediately after the ischaemia and daily for the 10 days' reperfusion; the oxygen group were given the same concentration of oxygen alone (13 mg/kg). Biochemical parameters fructosamine, phospholipase A2, catalase, superoxide dismutase and thiobarbituric acid reactive substances were measured, as well as renal plasma flow and glomerular filtration rate. KEY FINDINGS: Renal plasma flow and glomerular filtration rate decreased significantly in the positive controls and the oxygen group whereas values in the ozone group were similar to those in the negative control group. With respect to the biochemical parameters, ozone maintained a homeostasis redox, with significant increases in catalase and superoxide dismutase activities and similar values for phospholipase A2 and fructosamine compared with the negative control group. Fewer morphological alterations were seen in kidneys from the ozone group. No advantages were obtained in the positive control and oxygen groups. CONCLUSIONS: The protective effect of ozone may be explained by upregulation of the antioxidant defence system and beneficial effects on blood circulation and in oxygen metabolism. Ozone treatment may represent a therapeutic approach for minimising renal damage after transplantation.
Assuntos
Injúria Renal Aguda/tratamento farmacológico , Oxigênio/uso terapêutico , Ozônio/uso terapêutico , Injúria Renal Aguda/diagnóstico , Injúria Renal Aguda/fisiopatologia , Animais , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Modelos Animais de Doenças , Sequestradores de Radicais Livres/antagonistas & inibidores , Sequestradores de Radicais Livres/metabolismo , Frutosamina/metabolismo , Rim/efeitos dos fármacos , Testes de Função Renal , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Ozônio/análise , Fosfolipases A2/metabolismo , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/farmacologia , Espécies Reativas de Oxigênio/uso terapêutico , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico , Isquemia QuenteRESUMO
Apoptosis seems to play an important role in cancer immunotherapy outcome. We have studied the kinetic pattern of apoptosis induction in H125 human lung carcinoma xenografts after treatment with the monoclonal antibody (MAb) anti-epidermal growth factor receptor (EGFr) IOR EGF/r3. Tumor-bearing nude mice were injected intravenously with a single 8 mg/kg dose of IOR EGF/r3 and tumor specimens were taken up to 30 days post treatment. Apoptosis was measured by morphometric analysis of the histological sections at each tumor specimen over time points. The results showed a significant apoptotic response in tumors within six days after injection of this MAb reaching a peak at 20 days post treatment. The kinetics were very broad, with apoptotic cells present over the entire time-frame. However, the time course of the apoptotic index showed a significant difference to the mitotic index. Finally, the MAb-induced apoptosis was related to tumor growth delay indicating a probable arrest of cell cycle and a corresponding inhibition of tumor progression, which was corroborated by the Ki67 and proliferating cell nuclear antigen (PCNA) biomarkers.
Assuntos
Anticorpos Monoclonais/farmacologia , Apoptose/efeitos dos fármacos , Receptores ErbB/imunologia , Neoplasias/patologia , Animais , Humanos , Masculino , Camundongos , Camundongos NusRESUMO
In vivo preventive effects of a Mangifera indica L extract (Vimang) or its major component mangiferin on iron overload injury have been studied in rats given respectively, 50, 100, 250 mg kg(-1) body weight of Vimang, or 40 mg kg(-1) body weight of mangiferin, for 7 days prior to, and for 7 days following the administration of toxic amounts of iron-dextran. Both Vimang or mangiferin treatment prevented iron overload in serum as well as liver oxidative stress, decreased serum and liver lipid peroxidation, serum GPx activity, and increased serum and liver GSH, serum SOD and the animals overall antioxidant condition. Serum iron concentration was decreased although at higher doses, Vimang tended to increase it; percent tranferrin saturation, liver weight/body mass ratios, liver iron content was decreased. Treatment increased serum iron-binding capacity and decreased serum levels of aspartate-amine transferase (ASAT) and alanine-amine transferase (ALAT), as well as the number of abnormal Kupffer cells in iron-loaded livers. It is suggested that besides acting as antioxidants, Vimang extract or its mangiferin component decrease liver iron by increasing its excretion. Complementing earlier in vitro results from our group, it appears possible to support the hypothesis that Vimang and mangiferin present therapeutically useful effects in iron overload related diseases.
Assuntos
Antioxidantes/farmacologia , Sobrecarga de Ferro/tratamento farmacológico , Fígado/efeitos dos fármacos , Extratos Vegetais/farmacologia , Xantonas/farmacologia , Animais , Feminino , Glutationa/análise , Glutationa Peroxidase/sangue , Ferro/sangue , Sobrecarga de Ferro/sangue , Fígado/metabolismo , Fígado/patologia , Mangifera , Ratos , Ratos Wistar , Superóxido Dismutase/sangueRESUMO
This research was performed in order to determine the potential protective effects of ozonized sunflower oil (OSO) in the injury of rat gastric mucosa induced by absolute ethanol and as well as to elucidate the role of reactive oxygen species (ROS), lipid peroxidation, and some important constituents of antioxidant defense such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in these effects. OSO was administered to rats intragastrically by a cannula and it was applied during four days to animals. The doses of OSO administered daily to each group of rats were 4, 12, and 24 mg/kg, respectively, and one hour after the last treatment, absolute ethanol (1 mL/200 mg body weight) was administered. Our results showed that gastric ulcer index was significantly reduced in rats pretreated with OSO as compared with ethanol-treated controls. However, in rats pretreated with OSO, no significant reduction of TBARS content in gastric mucosa was found as compared to those rats treated with ethanol alone. In contrast, SOD and GSH-Px activities were significantly increased in gastric mucosa of OSO-pretreated rats with respect to those treated with ethanol alone. In summary, our results demonstrate that OSO pretreatment exerts protective effects in ethanol-induced gastric ulcers in rats. Furthermore, these results provide evidence that these protective effects of OSO are mediated at least partially by stimulation of some important antioxidant enzymes such as SOD and GSH-Px, which are scavengers of ROS and therefore prevent gastric injury induced by them.
Assuntos
Antioxidantes/farmacologia , Óleos de Plantas/farmacologia , Úlcera Gástrica/prevenção & controle , Animais , Catalase/metabolismo , Etanol , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patologia , Glutationa Peroxidase/metabolismo , Masculino , Ozônio/química , Óleos de Plantas/química , Ratos , Ratos Sprague-Dawley , Úlcera Gástrica/induzido quimicamente , Úlcera Gástrica/metabolismo , Óleo de Girassol , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
The objective of this work was the generation of an animal model of the SCA2 disease for future studies on the benefits of therapeutic molecules and neuropathological mechanisms that underline this human disorder. The transgenic fragment was microinjected into pronuclei of B6D2F1 X OF1 mouse hybrid strain. For Northern blots, RNAs were hybridized with a human cDNA fragment from the SCA2 gene and a mouse beta-actin cDNA fragment. Monoclonal antibody directed to the N-terminal of the ataxin 2 protein with 22Q was used for Western blot analysis. A rotating rod apparatus was utilized to measure motor coordination of mice. Immunohistochemical detection of Purkinje neurons was performed with anti-calbindin 28K as primary antibody. Ubiquitous expression of the SCA2 transgene with 75 CAG repeats regulated by the SCA2 self promoter was obtained after generation of our transgenic mice. Analysis of transgenic mice revealed significant differences of motor coordination compared with the wild type littermates. Specific degeneration of Purkinje neurons and transgene over-expression in the brain, liver and skeletal muscle, rather than in lungs and kidneys was also observed, resembling the expression pattern of the ataxin 2 in humans.
Assuntos
Proteínas do Tecido Nervoso/metabolismo , Regiões Promotoras Genéticas/fisiologia , Células de Purkinje/patologia , Degenerações Espinocerebelares/metabolismo , Degenerações Espinocerebelares/patologia , Análise de Variância , Animais , Ataxinas , Northern Blotting/métodos , Western Blotting/métodos , Calbindina 1 , Calbindinas , Modelos Animais de Doenças , Humanos , Imuno-Histoquímica/métodos , Camundongos , Camundongos Transgênicos , Atividade Motora/fisiologia , Proteínas do Tecido Nervoso/genética , Células de Purkinje/metabolismo , RNA Mensageiro/biossíntese , Sequências Reguladoras de Ácido Nucleico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Teste de Desempenho do Rota-Rod/métodos , Proteína G de Ligação ao Cálcio S100/metabolismo , Degenerações Espinocerebelares/fisiopatologia , Fatores de TempoRESUMO
Cisplatin (CDDP), an anticancer drug, induces remarkable toxicity in the kidneys of animals and humans and it has been well documented that reactive oxygen species and the renal antioxidant system are strongly involved in acute renal damage induced by CDDP. The aim of the present study was to investigate whether or not the renal antioxidant system plays also an important role in chronic renal damage induced by repeated doses of CDDP (1 mg/kg intraperitoneally twice weekly during 10 weeks in rats). In order to elucidate it, serum creatinine and urea levels, renal glutathione and thiobarbituric acid-reactive substances (TBARS) content, as well as renal superoxide dismutase and glutathione peroxidase activities were measured in the kidney homogenates of chronically CDDP-treated rats and additionally histological studies were performed in the rat kidneys. The chronic treatment with CDDP induced a significant increase in creatinine and urea levels in serum, but the other parameters mentioned above were not significantly modified as compared to the values in nontreated rats. Taking into account these results, we conclude that chronic CDDP administration induces also severe nephrotoxicity, in contrast to CDDP acute application, without any significant modification in the activity of relevant antioxidant enzymes such as superoxide dismutase and glutathione peroxidase, renal glutathione and lipid peroxides, by which the role of the antioxidant system in chronic nephrotoxicity induced by CDDP in rats is uncertain.
Assuntos
Antineoplásicos/toxicidade , Antioxidantes/metabolismo , Cisplatino/toxicidade , Nefropatias/induzido quimicamente , Rim/efeitos dos fármacos , Peróxidos Lipídicos/metabolismo , Animais , Antineoplásicos/administração & dosagem , Cisplatino/administração & dosagem , Glutationa/metabolismo , Humanos , Rim/citologia , Rim/metabolismo , Rim/patologia , Nefropatias/metabolismo , Nefropatias/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Extratos de Tecidos/química , Ureia/sangueRESUMO
The liver is damaged by sustained ischaemia during liver transplantation, and the reperfusion after ischaemia results in further functional impairment. Ozone oxidative preconditioning (OzoneOP) protected the liver against ischaemia/reperfusion (I/R) injury through different mechanisms. The aim of this study was to investigate the influence of the inhibition of protein synthesis on the protective actions conferred by OzoneOP in hepatic I/R. Rats were treated with cycloheximide (CHX) in order to promote protein synthesis inhibition after OzoneOP treatment. Plasma transaminases, malondialdehyde and 4-hydroxyalkenals and morphological characteristics were measured as an index of hepatocellular damage; Cu/Zn-superoxide dismutase (SOD), Mn-SOD, catalase, total hydroperoxides and glutathione levels as markers of endogenous antioxidant system. OzoneOP increased Mn-SOD isoform and ameliorated mitochondrial damage. CHX abrogated the protection conferred by OzonoOP and decreased Mn-SOD activity. Cellular redox balance disappeared when CHX was introduced. Protein synthesis is involved in the protective mechanisms mediated by OzoneOP. Ozone treatment preserved mitochondrial functions and cellular redox balance.
Assuntos
Transplante de Fígado , Fígado/lesões , Fígado/metabolismo , Ozônio/administração & dosagem , Biossíntese de Proteínas , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Catalase/metabolismo , Cicloeximida/farmacologia , Glutationa/metabolismo , Dissulfeto de Glutationa/metabolismo , Precondicionamento Isquêmico , Fígado/efeitos dos fármacos , Fígado/ultraestrutura , Transplante de Fígado/efeitos adversos , Transplante de Fígado/fisiologia , Masculino , Microscopia Eletrônica , Oxirredução , Inibidores da Síntese de Proteínas/farmacologia , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismoRESUMO
Increased formation of MG (methylglyoxal) and related protein glycation in diabetes has been linked to the development of diabetic vascular complications. Diabetes is also associated with impaired wound healing. In the present study, we investigated if prolonged exposure of rats to MG (50-75 mg/kg of body weight) induced impairment of wound healing and diabetes-like vascular damage. MG treatment arrested growth, increased serum creatinine, induced hypercholesterolaemia (all P < 0.05) and impaired vasodilation (P < 0.01) compared with saline controls. Degenerative changes in cutaneous microvessels with loss of endothelial cells, basement membrane thickening and luminal occlusion were also detected. Acute granulation appeared immature (P < 0.01) and was associated with an impaired infiltration of regenerative cells with reduced proliferative rates (P < 0.01). Immunohistochemical staining indicated the presence of AGEs (advanced glycation end-products) in vascular structures, cutaneous tissue and peripheral nerve fibres. Expression of RAGE (receptor for AGEs) appeared to be increased in the cutaneous vasculature. There were also pro-inflammatory and profibrotic responses, including increased IL-1beta (interleukin-1beta) expression in intact epidermis, TNF-alpha (tumour necrosis factor-alpha) in regions of angiogenesis, CTGF (connective tissue growth factor) in medial layers of arteries, and TGF-beta (transforming growth factor-beta) in glomerular tufts, tubular epithelial cells and interstitial endothelial cells. We conclude that exposure to increased MG in vivo is associated with the onset of microvascular damage and other diabetes-like complications within a normoglycaemic context.
Assuntos
Angiopatias Diabéticas/induzido quimicamente , Aldeído Pirúvico/farmacologia , Pele/lesões , Cicatrização , Animais , Glicemia/análise , Colesterol/sangue , Angiopatias Diabéticas/imunologia , Angiopatias Diabéticas/fisiopatologia , Modelos Animais de Doenças , Frutosamina/sangue , Injeções Intraperitoneais , Interleucina-1/análise , Masculino , Microcirculação , Neovascularização Patológica , Distribuição Aleatória , Ratos , Ratos Wistar , Pele/irrigação sanguínea , Pele/imunologia , Triglicerídeos/sangue , Fator de Necrose Tumoral alfa/análise , Vasodilatação/efeitos dos fármacosRESUMO
The h-R3 is a humanized monoclonal antibody (Mab) that binds to the external domain of the human epidermal growth factor receptor (EGF-R). It has being used for the treatment of head and neck tumors. Since it is known that different animal species have different EGF-R amino acid sequences, it raises the handicap that, a priori, the animal species relevant for the pharmacodynamic, pharmacokinetic, and toxicologic studies of this Mab are unknown. To elucidate relevant laboratory animal species, the authors investigated the immunohistochemical recognition by h-R3 Mab of EGF-R in small skin biopsy samples obtained from NMRI nu/nu, C57Bl/6, and OF-1 mice; Sprague-Dawley rats; Hartley guinea pigs; New Zealand rabbits; and Cercopithecus aethiops and Macaca fascicularis monkeys. Additionally, three human skin biopsies were obtained from trauma victims. The immunolocalization of EGF-R in different tissue sections was performed using the avidin-biotin peroxidase complex (ABC) technique. The skin sections from different tissues were tested with the biotinylated h-R3 Mab or with a biotinylated irrelevant Mab, used as negative control. The staining intensity was qualified as:-, no staining; +, weak staining; ++, moderate staining; +++, strong staining. Macaca fascicularis monkey, New Zealand rabbit, and OF-1 mouse skins had a strong staining intensity; Cercopithecus aethiops monkey and Sprague-Dawley rat skins showed a moderate to strong staining intensity; C57Bl/6 mouse skins showed a moderate staining intensity. No staining was observed in the skins from Hartley guinea pigs and MNRI nu/nu mice. The fact that h-R3 Mab recognizes other animal EGF receptors in addition to the human EGF-R makes it possible to perform relevant pharmacodynamic, pharmacokinetic, and toxicologic studies in some laboratory animals.
Assuntos
Anticorpos Monoclonais/imunologia , Receptores ErbB/análise , Pele/química , Animais , Chlorocebus aethiops , Receptores ErbB/imunologia , Receptores ErbB/metabolismo , Cobaias , Humanos , Imuno-Histoquímica , Macaca fascicularis , Camundongos , Camundongos Nus , Coelhos , Ratos , Ratos Sprague-Dawley , Pele/imunologia , Pele/metabolismoRESUMO
Se estudió el efecto del ozono por vía rectal a diferentes dosis sobre variables de función renal y la presión arterial sistólica en un modelo de glomerulonefritis tóxica experimental por adriamicina. La glomerulonefritis experimental es causa de insuficiencia renal, caracterizada por un daño renal progresivo. Existen diferentes esquemas de tratamiento, los cuales son muy caros y producen inmunosupresión, que afecta la calidad de vida del paciente. Para desarrollar este trabajo se utilizaron 40 ratas hembras Wistar de 200 g de peso, divididas en 4 grupos, uno control, otro control positivo, que recibieron adriamicina durante un período de 10 semanas, y otros 2 que recibieron terapia con ozono. A todos se les determinó la proteinuria, presión arterial sistólica y diuresis de 24 h. Los resultados mostraron que la terapia con ozono a una dosis de 0,3 mg/kg tuvo efecto renoprotector
Assuntos
Animais , Ratos , Doxorrubicina , Glomerulonefrite , Modelos Animais , Ozônio , Ratos Wistar , Insuficiência RenalRESUMO
Metabolism of arachidonic acid by cyclooxygenase is one of the primary sources of reactive oxygen species in the ischemic brain. Neuronal overexpression of cyclooxygenase-2 has recently been shown to contribute to neurodegeneration following ischemic injury. In the present study, we examined the possibility that the neuroprotective effects of the cyclooxygenase-2 inhibitor nimesulide would depend upon reduction of oxidative stress following cerebral ischemia. Gerbils were subjected to 5 min of transient global cerebral ischemia followed by 48 h of reperfusion and markers of oxidative stress were measured in hippocampus of gerbils receiving vehicle or nimesulide treatment at three different clinically relevant doses (3, 6 or 12 mg/kg). Compared with vehicle, nimesulide significantly (P<0.05) reduced hippocampal glutathione depletion and lipid peroxidation, as assessed by the levels of malondialdehyde (MDA), 4-hydroxy-alkenals (4-HDA) and lipid hydroperoxides levels, even when the treatment was delayed until 6 h after ischemia. Biochemical evidences of nimesulide neuroprotection were supported by histofluorescence findings using the novel marker of neuronal degeneration Fluoro-Jade B. Few Fluoro-Jade B positive cells were seen in CA1 region of hippocampus in ischemic animals treated with nimesulide compared with vehicle. These results suggest that nimesulide may protect neurons by attenuating oxidative stress and reperfusion injury following the ischemic insult with a wide therapeutic window of protection.