RESUMO
Clinical reports suggest close interactions between stressors, particularly those of long duration, and liver diseases, such as hepatic inflammation, that is proposed to occur via reactive oxygen species. In the present study we have used 21-day social isolation of male Wistar rats as a model of chronic stress to investigate protein expression/activity of liver antioxidant enzymes: superoxide dismutases (SODs), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GLR), and protein expression of their upstream regulators: glucocorticoid receptor (GR) and nuclear factor kappa B (NFkB). We have also characterized these parameters in either naive or chronically stressed animals that were challenged by 30-min acute immobilization. We found that chronic isolation caused decrease in serum corticosterone (CORT) and blood glucose (GLU), increase in NFkB signaling, and disproportion between CuZnSOD, peroxidases (CAT, GPx) and GLR, thus promoting H2O2 accumulation and prooxidative state in liver. The overall results suggested that chronic stress exaggerated responsiveness to subsequent stressor at the level of CORT and GLU, and potentiated GLR response, but compromised the restoration of oxido-reductive balance due to irreversible alterations in MnSOD and GPx.
Assuntos
Catalase/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Fígado/enzimologia , Estresse Fisiológico/fisiologia , Superóxido Dismutase/metabolismo , Doença Aguda , Animais , Glicemia/metabolismo , Doença Crônica , Corticosterona/sangue , Peróxido de Hidrogênio/metabolismo , Masculino , NF-kappa B/metabolismo , Ratos , Ratos Wistar , Receptores de Glucocorticoides/metabolismo , Transdução de Sinais/fisiologia , Superóxido Dismutase-1 , Glutationa Peroxidase GPX1RESUMO
In 2012, alcohol liver disease resulted in 3.3 million-5.9% of global deaths. This study introduced whey protection capacity against chronic alcohol-induced liver injury. Rats were orally administered to 12% ethanol solution in water (ad libitum, average 8.14 g of ethanol/kg body weight (b.w.)/day) alone or combined with whey ( per os, 2 g/kg b.w./day). After 6-week treatment, chronic ethanol consumption induced significant histopathological liver changes: congestion, central vein dilation, hepatic portal vein branch dilation, Kupffer cells hyperplasia, fatty liver changes, and hepatocytes focal necrosis. Ethanol significantly increased liver catalase activity and glutathione reductase protein expression without significant effects on antioxidative enzymes: glutathione peroxidase (GPx), copper-zinc-containing superoxide dismutase (CuZnSOD) and manganese-containing superoxide dismutase (MnSOD). Co-treatment with whey significantly attenuated pathohistological changes induced by ethanol ingestion and increased GSH-Px and nuclear factor kappa B (NF-κB) protein expression. Our results showed positive effects of whey on liver chronically exposed to ethanol, which seem to be associated with NF-κB-GPx signaling.
Assuntos
Doença Hepática Crônica Induzida por Substâncias e Drogas/tratamento farmacológico , Hepatopatias Alcoólicas/tratamento farmacológico , Substâncias Protetoras/uso terapêutico , Soro do Leite , Consumo de Bebidas Alcoólicas , Animais , Doença Hepática Crônica Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Crônica Induzida por Substâncias e Drogas/patologia , Glutationa Peroxidase/metabolismo , Fígado/efeitos dos fármacos , Fígado/patologia , Hepatopatias Alcoólicas/metabolismo , Hepatopatias Alcoólicas/patologia , Masculino , NF-kappa B/metabolismo , Substâncias Protetoras/farmacologia , Ratos WistarRESUMO
Gamma-irradiation (gamma-IR) is extensively used in the treatment of hormone-resistant prostate carcinoma. The objective of the present study was to investigate the effects of 60Co gamma-IR on the growth, cell cycle arrest and cell death of the human prostate cancer cell line DU 145. The viability of DU 145 cells was measured by the Trypan blue exclusion assay and the 3(4,5-dimethylthiazol-2-yl)-2,5,diphenyltetrazolium bromide test. Bromodeoxyuridine incorporation was used for the determination of cell proliferation. Cell cycle arrest and cell death were analyzed by flow cytometry. Superoxide dismutase (SOD), specifically CuZnSOD and MnSOD protein expression, after 10 Gy gamma-IR, was determined by Western immunoblotting analysis. Gamma-IR treatment had a significant (P < 0.001) antiproliferative and cytotoxic effect on DU 145 cells. Both effects were time and dose dependent. Also, the dose of gamma-IR which inhibited DNA synthesis and cell proliferation by 50% was 9.7 Gy. Furthermore, gamma-IR induced cell cycle arrest in the G2/M phase and the percentage of cells in the G2/M phase was increased from 15% (control) to 49% (IR cells), with a nonsignificant induction of apoptosis. Treatment with 10 Gy gamma-IR for 24, 48, and 72 h stimulated CuZnSOD and MnSOD protein expression in a time-dependent manner, approximately by 3- to 3.5-fold. These data suggest that CuZnSOD and MnSOD enzymes may play an important role in the gamma-IR-induced changes in DU 145 cell growth, cell cycle arrest and cell death.
Assuntos
Apoptose/efeitos da radiação , Ciclo Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Raios gama , Neoplasias da Próstata/patologia , Superóxido Dismutase/efeitos da radiação , Western Blotting , Linhagem Celular Tumoral/efeitos da radiação , Humanos , Masculino , Neoplasias da Próstata/metabolismoRESUMO
PURPOSE: Radiation treatment of breast cancer (BC) often results in post-therapy complications. The undesired sequelae could be avoided by the diagnostic screening of biomarkers for prediction of ionizing radiation (IR)-linked injury of healthy tissues. PATIENTS AND METHODS: The expression of antioxidative defence enzymes CuZn- and Mn-superoxide dismutase (CuZnSOD, MnSOD) and tumor suppressor protein p53 was measured in blood cells of 19 women with BC (age groups 30-45 and 46-60 years) and respective controls. The proteins were detected by specific immunostaining and quantified by laser-scanning densitometry. RESULTS: Constitutive expression of CuZnSOD was significantly elevated in the group of BC patients (up to 254 arbitrary units, AU/mL) relative to the control group (105-130 AU/mL). The constitutive expression of MnSOD was elevated (up to 94 AU/mL) in the group of BC patients relative to the controls (53-56 AU/mL). p53 was also constitutively more expressed in BC patients (35-42 AU/mL) than in controls (32-33 AU/mL). Both MnSOD and p53 were inducible by (60)Co gamma-ray IR (up to 170 AU/mL and 51 AU/mL, respectively) in the BC patient group. The levels of IR-induced p53 correlated inversely with MnSOD levels. CONCLUSION: The constitutive expression of all 3 proteins could be a useful biomarker for the presence of BC, but only MnSOD overexpression may be the predictive biomarker for selection of BC patients that would be less susceptible to IR-linked complications.
RESUMO
Gamma-irradiation (gamma-IR) is extensively used in the treatment of hormone-resistant prostate carcinoma. The objective of the present study was to investigate the effects of 60Co gamma-IR on the growth, cell cycle arrest and cell death of the human prostate cancer cell line DU 145. The viability of DU 145 cells was measured by the Trypan blue exclusion assay and the 3(4,5-dimethylthiazol-2-yl)-2,5,diphenyltetrazolium bromide test. Bromodeoxyuridine incorporation was used for the determination of cell proliferation. Cell cycle arrest and cell death were analyzed by flow cytometry. Superoxide dismutase (SOD), specifically CuZnSOD and MnSOD protein expression, after 10 Gy gamma-IR, was determined by Western immunoblotting analysis. gamma-IR treatment had a significant (P < 0.001) antiproliferative and cytotoxic effect on DU 145 cells. Both effects were time and dose dependent. Also, the dose of gamma-IR which inhibited DNA synthesis and cell proliferation by 50 percent was 9.7 Gy. Furthermore, gamma-IR induced cell cycle arrest in the G2/M phase and the percentage of cells in the G2/M phase was increased from 15 percent (control) to 49 percent (IR cells), with a nonsignificant induction of apoptosis. Treatment with 10 Gy gamma-IR for 24, 48, and 72 h stimulated CuZnSOD and MnSOD protein expression in a time-dependent manner, approximately by 3- to 3.5-fold. These data suggest that CuZnSOD and MnSOD enzymes may play an important role in the gamma-IR-induced changes in DU 145 cell growth, cell cycle arrest and cell death.