RESUMO
Liver regeneration is critical to survival after traumatic injuries, exposure to hepatotoxins, or surgical interventions, yet the underlying signaling and metabolic pathways remain unclear. In this study, we show that hepatocyte-specific loss of the mitochondrial deacetylase SIRT3 drastically impairs regeneration and worsens mitochondrial function after partial hepatectomy. Sirtuins, including SIRT3, require NAD as a cosubstrate. We previously showed that the NAD precursor nicotinamide riboside (NR) promotes liver regeneration, but whether this involves sirtuins has not been tested. Here, we show that despite their NAD dependence and critical roles in regeneration, neither SIRT3 nor its nuclear counterpart SIRT1 is required for NR to enhance liver regeneration. NR improves mitochondrial respiration in regenerating WT or mutant livers and rapidly increases oxygen consumption and glucose output in cultured hepatocytes. Our data support a direct enhancement of mitochondrial redox metabolism as the mechanism mediating improved liver regeneration after NAD supplementation and exclude signaling via SIRT1 and SIRT3. Therefore, we provide the first evidence to our knowledge for an essential role for a mitochondrial sirtuin during liver regeneration and insight into the beneficial effects of NR.
Assuntos
Regeneração Hepática/fisiologia , Mitocôndrias Hepáticas/fisiologia , Niacinamida/análogos & derivados , Compostos de Piridínio/farmacologia , Sirtuína 3/metabolismo , Animais , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Regeneração Hepática/efeitos dos fármacos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Mitocôndrias Hepáticas/efeitos dos fármacos , Niacinamida/farmacologia , Oxirredução , Sirtuína 1/genética , Sirtuína 1/metabolismo , Sirtuína 3/genéticaRESUMO
Nonalcoholic fatty liver disease (NAFLD) is quickly becoming the most common liver disease worldwide. Within the NAFLD spectrum, patients with nonalcoholic steatohepatitis (NASH) are at the highest risk of developing cirrhosis and disease progression to hepatocellular carcinoma. To date, therapeutic options for NASH patients have been ineffective, and therefore, new options are urgently needed. Hence, a model system to develop new therapeutic interventions is needed. Here, we introduce two new in vitro models of steatosis induction in HepG2 cells and primary murine hepatocytes. We used a recently discovered novel class of bioactive anti-inflammatory lipids called branched fatty acid esters of hydroxyl fatty acids. Among these bioactive lipids, palmitic-acid-9-hydroxy-stearic-acid (9-PAHSA) is the most promising as a representative nondrug therapy based on dietary supplements or nutritional modifications. In this study, we show a therapeutic effect of 9-PAHSA on lipotoxicity in steatotic primary hepatocytes and HepG2 cells. This could be shown be increased viability and decreased steatosis. Furthermore, we could demonstrate a preventive effect in HepG2 cells. The outcome of 9-PAHSA administration is both preventative and therapeutically effective for hepatocytes with limited damage. In conclusion, bioactive lipids like 9-PAHSA offer new hope for prevention or treatment in patients with fatty liver and steatosis.
Assuntos
Fígado Gorduroso/patologia , Hepatócitos/efeitos dos fármacos , Mitocôndrias Hepáticas/efeitos dos fármacos , Doenças Mitocondriais/prevenção & controle , Ácido Palmítico/farmacologia , Ácidos Esteáricos/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Citoproteção/efeitos dos fármacos , Fígado Gorduroso/metabolismo , Fígado Gorduroso/fisiopatologia , Células Hep G2 , Hepatócitos/patologia , Hepatócitos/fisiologia , Humanos , Fígado/efeitos dos fármacos , Fígado/patologia , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias Hepáticas/fisiologia , Doenças Mitocondriais/etiologiaRESUMO
Maternal smoking leads to glucose and lipid metabolic disorders and hepatic damage in the offspring, potentially due to mitochondrial oxidative stress. Mitoquinone mesylate (MitoQ) is a mitochondrial targeted antioxidant with high bioavailability. This study aimed to examine the impact of maternal cigarette smoke exposure (SE) on offspring's metabolic profile and hepatic damage, and whether maternal MitoQ supplementation during gestation can affect these changes. Female Balb/c mice (eight weeks) were either exposed to air or SE for six weeks prior to mating and throughout gestation and lactation. A subset of the SE dams were supplied with MitoQ in the drinking water (500 µmol/L) during gestation and lactation. Intraperitoneal glucose tolerance test was performed in the male offspring at 12 weeks and the livers and plasma were collected at 13 weeks. Maternal SE induced glucose intolerance, hepatic steatosis, mitochondrial oxidative stress and related damage in the adult offspring. Maternal MitoQ supplementation reduced hepatic mitochondrial oxidative stress and improved markers of mitophagy and mitochondrial biogenesis. This may restore hepatic mitochondrial health and was associated with an amelioration of glucose intolerance, hepatic steatosis and pathological changes induced by maternal SE. MitoQ supplementation may potentially prevent metabolic dysfunction and hepatic pathology induced by intrauterine SE.
Assuntos
Fígado Gorduroso/induzido quimicamente , Exposição Materna , Síndrome Metabólica/induzido quimicamente , Compostos Organofosforados/farmacologia , Poluição por Fumaça de Tabaco/efeitos adversos , Ubiquinona/análogos & derivados , Animais , Antioxidantes/administração & dosagem , Antioxidantes/farmacologia , Feminino , Lactação , Lipidômica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mitocôndrias Hepáticas/fisiologia , Compostos Organofosforados/administração & dosagem , Estresse Oxidativo , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Ubiquinona/administração & dosagem , Ubiquinona/farmacologiaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Disturbed mitochondrial function and energy crisis serve as key mechanisms for the development of liver injury. Hence, targeting cellular mitochondria in liver diseases might serve as a therapeutic option. Tapinanthus globifer (A.Rich.) Tiegh. has been used in traditional medicine in the management of liver disease. However, there is no scientific evidence supporting such use. AIM OF THE STUDY: The current investigation was designed to evaluate the protective role of Tapinanthus globifer treatment on the liver mitochondrial function after the induction of hepatotoxicity by the hepatotoxic agent Fe2+in vitro. MATERIALS AND METHODS: In this study, isolated mitochondria from rats' liver was incubated with Fe2+ (10⯵M) for 1â¯h in the absence or presence of T. globifer (50, 100 and 200⯵g/mL) metanolic extract (MVA). Mitochondrial viability, mitochondrial membrane potential (ΔΨm), mitochondrial swelling (MPTP)., total thiol content, lipid peroxidation (TBARS) and reactive oxygen species (ROS) production were measured. HPLC-DAD was used to identify potential phytochemicals in MVA. RESULTS: (MVA) was able to improve mitochondrial dysfunction induced by Fe2+, by attenuating MTT reduction, increased ΔΨm and mitochondrial swelling. Reduced total thiol and non-protein thiol contents which were associated with increased lipid peroxidation and ROS generation in Fe2+-treated mitochondria were significantly improved by MVA co-treatment. HPLC-DAD analysis revealed the presence of gallic acid, catechin, epigallocatechin, caffeic acid, rutin, glycoside flavonoid and quercetin in MVA that can be responsible for its beneficial effect. CONCLUSION: MVA phyto-compounds enhance mitochondrial redox signaling and possess mitochondrial function improving potential, thereby, providing scientific basis for its use in traditional medicine.
Assuntos
Loranthaceae , Mitocôndrias Hepáticas/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Ferro/toxicidade , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias Hepáticas/fisiologia , Dilatação Mitocondrial/efeitos dos fármacos , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/química , Folhas de Planta , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismoRESUMO
OBJECTIVES: Acquired aplastic anemia (AA) is a hematopoietic stem cell disease that leads to hematopoietic disorder and peripheral blood pancytopenia. We investigated whether nutritional support is helpful to AA recovery. METHODS: We established a rat model with AA. A nutrient mixture was administered to rats with AA through different dose gavage once per day for 55 d. Animals in this study were assigned to one of five groups: normal control (NC; group includes normal rats); AA (rats with AA); high dose (AA + nutritional mixture, 2266.95 mg/kg/d); medium dose (1511.3 mg/kg/d); and low dose (1057.91 mg/kg/d). The effects of nutrition administration on general status and mitochondrial function of rats with AA were evaluated. RESULTS: The nutrient mixture with which the rats were supplemented significantly improved weight, peripheral blood parameters, and histologic parameters of rats with AA in a dose-dependent manner. Furthermore, we observed that the number of mitochondria in the liver, spleen, kidney, and brain was increased after supplementation by transmission electron microscopy analysis. Nutrient administration also improved mitochondrial DNA content, adenosine triphosphate content, and membrane potential but inhibited oxidative stress, thus, repairing the mitochondrial dysfunction of the rats with AA. CONCLUSIONS: Taken together, nutrition supplements may contribute to the improvement of mitochondrial function and play an important role in the recuperation of rats with AA.
Assuntos
Anemia Aplástica/fisiopatologia , Anemia Aplástica/terapia , Mitocôndrias/fisiologia , Apoio Nutricional/métodos , Trifosfato de Adenosina/análise , Anemia Aplástica/patologia , Animais , Encéfalo/ultraestrutura , DNA/análise , Modelos Animais de Doenças , Rim/ultraestrutura , Potencial da Membrana Mitocondrial/fisiologia , Microscopia Eletrônica de Transmissão , Mitocôndrias/química , Mitocôndrias/patologia , Mitocôndrias Hepáticas/patologia , Mitocôndrias Hepáticas/fisiologia , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Baço/ultraestruturaRESUMO
Moutan Cortex, the root bark of Paeonia suffruticosa ANDREWS in Ranunculaceae, has widely demonstrated analgesic, anti-spasmodic, and anti-inflammatory effects in various cancer and immune cell lines. Oxidative stress is associated with development of several diseases, including liver disease. We prepared the water extract of Moutan Cortex (MCE) to investigate the cytoprotective activities and its mechanism. MCE protected hepatocytes from arachidonic acid (AA)+iron induced oxidative stress, as indicated by reactive oxygen species (ROS) production and cell viability analysis. MCE also suppressed mitochondrial dysfunction in AA+iron-treated human hepatocyte-derived hepatocellular carcinoma cell line, HepG2 cells. In addition, MCE treatment induces AMP-activated protein kinase (AMPK) and liver kinase B1 phosphorylation, which play a role in inhibition of oxidative stress induced cell death. Moreover, one of the MCE compounds, chlorogenic acid, exerted protective effects against oxidative stress and apoptosis. Taken together, MCE protected hepatocytes against AA+iron-induced oxidative stress through AMPK activation, and may be a candidate for the treatment of liver disease.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Hepatócitos/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Quinases Proteína-Quinases Ativadas por AMP , Proteínas Quinases Ativadas por AMP/metabolismo , Ácido Araquidônico , Linhagem Celular Tumoral , Ácido Clorogênico/farmacologia , Glutationa/metabolismo , Hepatócitos/metabolismo , Humanos , Peróxido de Hidrogênio/metabolismo , Ferro , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias Hepáticas/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Paeonia , Proteínas Serina-Treonina Quinases/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
Celastrol is a natural terpenoid isolated from Tripterygium wilfordii, a well-known Chinese medicinal herb that presents anti-proliferative activities in several cancer cell lines. Here, we investigated whether celastrol induces apoptosis on hepatocellular carcinoma Bel-7402 cells and further explored the underlying molecular mechanisms. Celastrol caused a dose- and time-dependent growth inhibition and apoptosis of Bel-7402 cells. It increased apoptosis through the up-regulation of Bax and the down-regulation of Bcl-2 in Bel-7402 cells. Moreover, celastrol induced the release of cytochrome c and increased the activation of caspase-3 and caspase-9, suggesting that celastrol-induced apoptosis was related to the mitochondrial pathway. These results indicated that celastrol could induce apoptosis in Bel-7402 cells, which may be associated with the activation of the mitochondria-mediated pathway.
Assuntos
Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Mitocôndrias Hepáticas/genética , Mitocôndrias Hepáticas/fisiologia , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Triterpenos/farmacologia , Carcinoma Hepatocelular/genética , Caspase 3/metabolismo , Caspase 9/metabolismo , Citocromos c/metabolismo , Relação Dose-Resposta a Droga , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Hepáticas/genética , Triterpenos Pentacíclicos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Tripterygium/química , Triterpenos/isolamento & purificação , Células Tumorais Cultivadas , Proteína X Associada a bcl-2/metabolismoRESUMO
We previously demonstrated that iron overload induces liver damage by causing the formation of reactive oxygen species (ROS). Taurine is a potent free radical scavenger that attenuates the damage caused by excessive oxygen free radicals. Therefore, the aim of the present study was to investigate whether taurine could reduce the hepatotoxicity of iron overload with regard to ROS production. Mice were intraperitoneally injected with iron 5 days/week for 13 weeks to achieve iron overload. It was found that iron overload resulted in liver dysfunction, increased apoptosis and elevated oxidative stress. Taurine supplementation increased liver taurine levels by 40% and led to improved liver function, as well as a reduction in apoptosis, ROS formation and mitochondrial swelling and an attenuation in the loss of the mitochondrial membrane potential. Treatment with taurine mediated a reduction in oxidative stress in ironoverloaded mice, attenuated liver lipid peroxidation, elevated antioxidant enzyme activities and maintained reduced glutathione levels. These results indicate that taurine reduces ironinduced hepatic oxidative stress, preserves liver function and inhibits hepatocyte apoptosis. Therefore, taurine may be a potential therapeutic drug to reduce liver damage caused by iron overload.
Assuntos
Sobrecarga de Ferro/tratamento farmacológico , Fígado/efeitos dos fármacos , Estresse Oxidativo , Substâncias Protetoras/administração & dosagem , Taurina/administração & dosagem , Alanina Transaminase/sangue , Animais , Apoptose/efeitos dos fármacos , Aspartato Aminotransferases/sangue , Catalase/metabolismo , Modelos Animais de Doenças , Glutationa Peroxidase/metabolismo , Hepatócitos/metabolismo , Peroxidação de Lipídeos , Fígado/enzimologia , Fígado/patologia , Masculino , Malondialdeído/sangue , Potencial da Membrana Mitocondrial , Camundongos , Mitocôndrias Hepáticas/fisiologia , Dilatação MitocondrialRESUMO
PURPOSE: To analyze the role of hyperbaric oxygen therapy as hepatic preconditioning in rats submitted to hepatic ischemia and reperfusion. METHODS: Wistar rats were randomly divided into three groups: SHAM, rats submitted to surgical stress without hepatic ischemia and reperfusion, I/R, rats submitted to total hepatic pedicle ischemia for 30 min, followed by 5 min of reperfusion; HBOI/R, rats submitted to 60 minutes of hyperbaric oxygen therapy at 2 atm and immediately submitted to the experimental protocol of ischemia and reperfusion. Liver function was assessed by measuring serum alanine aminotransferase and aspartate aminotransferase, as well as mitochondrial function by determining states 3 and 4 of mitochondrial respiration, respiratory control rate and mitochondrial permeability transition (mitochondrial swelling). The results were analyzed by the Mann-Whitney test and all P-values <0.05 were considered significant. RESULTS: There were significant differences in serum aspartate aminotransferase values in groups SHAM vs. HBOI/R, I/R vs HBOI/R, alanine aminotransferase in groups SHAM and I/R; State 3 in SHAM groups vs. I/R, SHAM vs. HBOI/R, State 4 in I/R vs HBOI/R groups, respiratory control rate in SHAM vs I/R groups; mitochondrial swelling in SHAM vs. I/R groups, and SHAM vs HBOI/R. CONCLUSION: Hyperbaric preconditioning improved hepatic mitochondrial function and decreased serum markers of liver injury in the ischemia and reperfusion process.
Assuntos
Oxigenoterapia Hiperbárica/métodos , Precondicionamento Isquêmico/métodos , Fígado/irrigação sanguínea , Mitocôndrias Hepáticas/fisiologia , Traumatismo por Reperfusão/prevenção & controle , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Respiração Celular , Fígado/fisiologia , Masculino , Consumo de Oxigênio , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
PURPOSE: To evaluate the effect of the hyperbaric oxygen (HBO) treatment as a pre-conditioning for I/R effects in the liver ischemia. METHODS: Fifty-seven male Wistar rats (260-300g) were submitted to the following procedures: SHAM; I/R, rats submitted to I/R, consisting of partial ischemia of 70% of the liver for 90 minutes followed by 15 minutes of reperfusion; HBO I/R 1 ATA, 30 minutes of HBO treatment at the pressure of 1 absolute atmosphere (ATA) during the ischemia time. HBO I/R 2 ATA, 30 minutes of HBO (2 ATA) during the ischemia time. Pre HBO I/R 30', rats submitted to 30 minutes of HBO (2 ATA) immediately before the I/R time. Pre HBO I/R 90', rats submitted to 90 minutes of HBO (2 ATA) immediately before the I/R time. RESULTS: There was a significant worsening of all the parameters of mitochondrial energy production (state 3, 4, RCR and Swelling) in the I/R group, when compared to the Sham group (I/R Assuntos
Oxigenoterapia Hiperbárica/métodos
, Precondicionamento Isquêmico/métodos
, Fígado/irrigação sanguínea
, Traumatismo por Reperfusão/terapia
, Alanina Transaminase/sangue
, Animais
, Aspartato Aminotransferases/sangue
, Respiração Celular
, Edema/etiologia
, Masculino
, Mitocôndrias Hepáticas/fisiologia
, Doenças Mitocondriais/etiologia
, Ratos Wistar
, Reprodutibilidade dos Testes
, Fatores de Tempo
, Resultado do Tratamento
RESUMO
PURPOSE: To determine the effect of hyperbaric hyperoxia as hepatic preconditioning on hepatocellular integrity in rats submitted to intermittent hepatic ischemia/reperfusion injury. METHODS: Twenty male Wistar rats were divided into 4 groups (SHAM, I/R, HBO-I/R and CONTROL). The surgical technique consisted of total clamping of the hepatic pedicle for 15 min, followed by reperfusion for 5 min, performed twice. The application of hyperbaric oxygen (HBO) was carried out in a collective chamber (simultaneous exposure of 4 rats) pressurized directly with oxygen at 2 ATA for 60 min. Tissue malondialdehyde (MDA) levels were determined and blood samples were collected for the determination of serum AST and ALT levels. Data were analyzed statistically by the Mann-Whitney test, with the level of significance set at p < 0.05. RESULTS: A statistically significant difference in MDA (p< 0.05) was observed between control and HBO-I/R, but not between control and I/R. Regarding AST, there was a difference between control and I/R and HBO-I/R. Analysis of ALT revealed a significant difference between control and I/R (p<0.05) and between I/R and HBO-I/R, with no difference between control and HBO-IR. CONCLUSION: Hyperoxic preconditioning proved to be favorable regarding alanine transaminase, but not aspartate aminotranserase or malondialdehyde levels.
Assuntos
Oxigenoterapia Hiperbárica/métodos , Precondicionamento Isquêmico/métodos , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/terapia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Masculino , Malondialdeído/análise , Mitocôndrias Hepáticas/fisiologia , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
PURPOSE: To evaluate the effects of hyperbaric oxygen on rats submitted to hepatic ischemia and reperfusion. METHODS: Twenty-three Wistar rats were divided at random into 3 groups: SHAM, rats submitted to surgical and anesthetic stress without induction of hepatic ischemia/reperfurion; I/R, rats submitted to total ischemia of the hepatic pedicle for 25 min followed by 5 min of reperfusion; HBOI/R, rats submitted to 60 min of hyperbaric oxygen therapy at a pressure of 2 absolute atmospheres immediately after the experimental protocol of ischemia/reperfusion. Hepatic function was evaluated by quantitation of serum alanine aminotranferase (ALT) and aspartate aminotransferase (AST), and by mitochondrial function through the determination of states 3 and 4 of mitochondrial respiration, respiratory control ratio (RCR) and mitochondrial swelling. Data were analyzed by the Mann-Whitney test, with the level of significance set at p <0.05. RESULTS: There was a significant difference in state 3 values for the SHAM group vs I/R and I/R vs IRHBO, in state 4 values for the SHAM group vs I/R; and in mitochondrial swelling for the SHAM groups vs I/RHBO, SHAM vs I/R, and IR vs I/RHBO. CONCLUSION: The use of hyperbaric oxygen after I/R improved in a relative manner both the production of energy and the effects on the mitochondrial wall.
Assuntos
Oxigenoterapia Hiperbárica/métodos , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/terapia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Respiração Celular , Masculino , Mitocôndrias Hepáticas/fisiologia , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
PURPOSE: To evaluate the effects of hyperbaric oxygen on rats submitted to hepatic ischemia and reperfusion. METHODS: Twenty-three Wistar rats were divided at random into 3 groups: SHAM, rats submitted to surgical and anesthetic stress without induction of hepatic ischemia/reperfurion; I/R, rats submitted to total ischemia of the hepatic pedicle for 25 min followed by 5 min of reperfusion; HBOI/R, rats submitted to 60 min of hyperbaric oxygen therapy at a pressure of 2 absolute atmospheres immediately after the experimental protocol of ischemia/reperfusion. Hepatic function was evaluated by quantitation of serum alanine aminotranferase (ALT) and aspartate aminotransferase (AST), and by mitochondrial function through the determination of states 3 and 4 of mitochondrial respiration, respiratory control ratio (RCR) and mitochondrial swelling. Data were analyzed by the Mann-Whitney test, with the level of significance set at p <0.05. RESULTS: There was a significant difference in state 3 values for the SHAM group vs I/R and I/R vs IRHBO, in state 4 values for the SHAM group vs I/R; and in mitochondrial swelling for the SHAM groups vs I/RHBO, SHAM vs I/R, and IR vs I/RHBO. CONCLUSION: The use of hyperbaric oxygen after I/R improved in a relative manner both the production of energy and the effects on the mitochondrial wall. .
Assuntos
Animais , Masculino , Oxigenoterapia Hiperbárica/métodos , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/terapia , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Respiração Celular , Mitocôndrias Hepáticas/fisiologia , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
PURPOSE: To determine the effect of hyperbaric hyperoxia as hepatic preconditioning on hepatocellular integrity in rats submitted to intermittent hepatic ischemia/reperfusion injury. METHODS: Twenty male Wistar rats were divided into 4 groups (SHAM, I/R, HBO-I/R and CONTROL). The surgical technique consisted of total clamping of the hepatic pedicle for 15 min, followed by reperfusion for 5 min, performed twice. The application of hyperbaric oxygen (HBO) was carried out in a collective chamber (simultaneous exposure of 4 rats) pressurized directly with oxygen at 2 ATA for 60 min. Tissue malondialdehyde (MDA) levels were determined and blood samples were collected for the determination of serum AST and ALT levels. Data were analyzed statistically by the Mann-Whitney test, with the level of significance set at p < 0.05. RESULTS: A statistically significant difference in MDA (p< 0.05) was observed between control and HBO-I/R, but not between control and I/R. Regarding AST, there was a difference between control and I/R and HBO-I/R. Analysis of ALT revealed a significant difference between control and I/R (p<0.05) and between I/R and HBO-I/R, with no difference between control and HBO-IR. CONCLUSION: Hyperoxic preconditioning proved to be favorable regarding alanine transaminase, but not aspartate aminotranserase or malondialdehyde levels. .
Assuntos
Animais , Masculino , Oxigenoterapia Hiperbárica/métodos , Precondicionamento Isquêmico/métodos , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/terapia , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Malondialdeído/análise , Mitocôndrias Hepáticas/fisiologia , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
PURPOSE: To evaluate the effect of the hyperbaric oxygen (HBO) treatment as a pre-conditioning for I/R effects in the liver ischemia. METHODS: Fifty-seven male Wistar rats (260-300g) were submitted to the following procedures: SHAM; I/R, rats submitted to I/R, consisting of partial ischemia of 70% of the liver for 90 minutes followed by 15 minutes of reperfusion; HBO I/R 1 ATA, 30 minutes of HBO treatment at the pressure of 1 absolute atmosphere (ATA) during the ischemia time. HBO I/R 2 ATA, 30 minutes of HBO (2 ATA) during the ischemia time. Pre HBO I/R 30', rats submitted to 30 minutes of HBO (2 ATA) immediately before the I/R time. Pre HBO I/R 90', rats submitted to 90 minutes of HBO (2 ATA) immediately before the I/R time. RESULTS: There was a significant worsening of all the parameters of mitochondrial energy production (state 3, 4, RCR and Swelling) in the I/R group, when compared to the Sham group (I/R <Sham, p<0.05). There was also a significant worsening in state 4, RCR and mitochondrial edema in the Pre HBO I/R 90' group compared to the I/R group. Hepatic enzyme concentrations were significantly higher in the I/R group. CONCLUSION: The use of hyperbaric oxygen before and during I/R did not improve the production of hepatocellular energy reduced by I/R, nor did it prevent the installation of mitochondrial edema induced by Iischemia/reperfusion. .
Assuntos
Animais , Masculino , Oxigenoterapia Hiperbárica/métodos , Precondicionamento Isquêmico/métodos , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/terapia , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Respiração Celular , Edema/etiologia , Mitocôndrias Hepáticas/fisiologia , Doenças Mitocondriais/etiologia , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
PURPOSE: To analyze the role of hyperbaric oxygen therapy as hepatic preconditioning in rats submitted to hepatic ischemia and reperfusion. METHODS: Wistar rats were randomly divided into three groups: SHAM, rats submitted to surgical stress without hepatic ischemia and reperfusion, I/R, rats submitted to total hepatic pedicle ischemia for 30 min, followed by 5 min of reperfusion; HBOI/R, rats submitted to 60 minutes of hyperbaric oxygen therapy at 2 atm and immediately submitted to the experimental protocol of ischemia and reperfusion. Liver function was assessed by measuring serum alanine aminotransferase and aspartate aminotransferase, as well as mitochondrial function by determining states 3 and 4 of mitochondrial respiration, respiratory control rate and mitochondrial permeability transition (mitochondrial swelling). The results were analyzed by the Mann-Whitney test and all P-values <0.05 were considered significant. RESULTS: There were significant differences in serum aspartate aminotransferase values in groups SHAM vs. HBOI/R, I/R vs HBOI/R, alanine aminotranferase in groups SHAM and I/R; State 3 in SHAM groups vs. I/R, SHAM vs. HBOI/R, State 4 in I/R vs HBOI/R groups, respiratory control rate in SHAM vs I/R groups; mitochondrial swelling in SHAM vs. I/R groups, and SHAM vs HBOI/R. CONCLUSION: Hyperbaric preconditioning improved hepatic mitochondrial function and decreased serum markers of liver injury in the ischemia and reperfusion process. .
Assuntos
Animais , Masculino , Oxigenoterapia Hiperbárica/métodos , Precondicionamento Isquêmico/métodos , Fígado/irrigação sanguínea , Mitocôndrias Hepáticas/fisiologia , Traumatismo por Reperfusão/prevenção & controle , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Respiração Celular , Fígado/fisiologia , Consumo de Oxigênio , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
Berberine is an isoquinoline alkaloid with anti-diabetic properties. Despite the central role of liver and thus hepatic mitochondria in whole-body metabolism, berberine effects on hepatic mitochondrial function in an obesity model are still unknown. Here, we demonstrate that berberine treatment recovers mitochondrial efficiency when altered by a high-fat feeding. Mitochondria isolated from the liver of high-fat fed rats exhibited decreased capacity to accumulate calcium and impaired oxidative phosphorylation (OXPHOS) capacity, as shown by impaired mitochondrial membrane potential, oxygen consumption and cellular ATP levels. Interestingly, the recovery of mitochondrial function by berberine was associated with an increased activity of the mitochondrial sirtuin 3 (SirT3). In conclusion, berberine potent protective effects against metabolic syndrome may rely on increasing mitochondrial SirT3 activity, normalizing mitochondrial function and preventing a state of energetic deficit caused by impaired OXPHOS.
Assuntos
Berberina/farmacologia , Dieta Hiperlipídica , Mitocôndrias Hepáticas/efeitos dos fármacos , Sirtuína 3/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Sequência de Bases , Berberina/administração & dosagem , Primers do DNA , Potencial da Membrana Mitocondrial , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias Hepáticas/fisiologia , Fosforilação Oxidativa , Ratos , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Sirtuína 3/genéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Liangxue Huayu Recipe (LHR) as a classical prescription is clinically employed to treat liver diseases in traditional Chinese medicine. AIM OF STUDY: In this study, we attempt to show that LHR attenuates hepatocyte apoptosis and hepatic injury induced by lipopolysaccharide (LPS) and D-galactosamine (GalN) in rats. The present study was also designed to examine whether LHR had the protective effects on d-GalN and tumor necrosis factor-α (TNF-α)-treated human L02 hepatocytes and its possible association with the mitochondrial pathway. MATERIALS AND METHODS: LHR is composed of three traditional Chinese medicines: Herba Rehmannia, Rhubarb and Radix Paeoniae Rubra. LHR at 541, 1082 and 2164 mg/kg was orally administered to model and normal rats for 7 days. The effects of LHR on serum levels of liver enzymes, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were measured. Hepatocyte apoptosis in vivo was assessed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method. Apoptosis in vitro and related morphological changes of human L02 hepatocytes were determined by high content screening (HCS) assay. The expression levels of Bcl-2, Bax and cytochrome c were detected by Western-blot analysis in L02 cells. In addition, the activities of caspase-3 and caspase-9 were tested by enzyme-linked immunosorbent detector. RESULTS: It revealed that LHR pretreatment effectively ameliorated the GalN/LPS-induced elevation of serum ALT and AST levels, and attenuated hepatocyte apoptosis in the rat model characterized by the addition of GalN/LPS. In subsequent experiments in vitro, LHR also attenuated GalN/TNF-α-induced apoptosis in human L02 hepatocytes. Furthermore, LHR improved the mitochondrial function, inhibited the upregulation of Bax/Bcl-2 protein ratio, decreased the release of cytochrome c from the mitochondria into the cytosol, as well as inhibited caspase-3 and caspase-9 activation in this cell model. CONCLUSIONS: These results indicate that LHR is effective in attenuating hepatocyte apoptosis both in vivo and in vitro, and this effect is partly mediated through the activation of the mitochondrial pathway and subsequent regulation of particular pro-apoptotic gene expression.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Fígado/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 9/metabolismo , Linhagem Celular , Citocromos c/metabolismo , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Fígado/metabolismo , Masculino , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/fisiologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Proteína X Associada a bcl-2/metabolismoRESUMO
PURPOSE: To analyze the effects of hyperbaric oxygen therapy on liver function in rats previously subjected to ischemia and reperfusion. METHODS: A randomly distribution of 23 Wistar rats was conducted into three groups: SHAM, animals subjected to surgical stress without restricting blood flow by clamping the hepatic pedicle, IR, rats underwent hepatic vascular occlusion intermittently for two complete cycles of 15 minutes of ischemia followed by 5 min of reperfusion, IR / HBO, rats underwent hepatic pedicle clamping and thereafter exposed to hyperbaric oxygen pressure of 2 absolute atmospheres for 60 minutes. We evaluated liver function through mitochondrial function, determined by the stages 3 and 4 of respiration, respiratory control ratio (RCR) and mitochondrial permeability transition (Swelling). Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also quantified . We analyzed the results using the Mann-Whitney test and were considered significant all results with p <0.05. RESULTS: There were significant differences between the results of stage 3 in SHAM vs IR group ; of the stage 4 in the groups IR vs SHAM and SHAM vs IR /HBO; of the Respiratory Control Ratio (RCR) in the group IR vs IR / HBO ; of alanine aminotransferase in the groups IR vs SHAM , SHAM vs IR/HBO and IR vs IR / HBO; aspartate aminotransferase in the groups SHAM vs IR and SHAM vs IR / HBO. CONCLUSION: The whole analysis of the mitochondrial function indicators permits us to conclude that the hyperbaric oxygen therapy acted as a protective agent of the mitochondrial function, minimizing the ischemia-reperfusion injury of the hepatic parenchyma.
Assuntos
Oxigenoterapia Hiperbárica , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/prevenção & controle , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Respiração Celular , Fígado/fisiopatologia , Masculino , Mitocôndrias , Mitocôndrias Hepáticas/fisiologia , Substâncias Protetoras , Distribuição Aleatória , Ratos , Ratos Wistar , Reperfusão , Traumatismo por Reperfusão/sangueRESUMO
PURPOSE: to assess the effect of hyperbaric oxygen (HBO) as pre-conditioning on periodic liver ischemia/reperfusion injury. METHODS: Thirty-six male Wistar rats were divided into 4 groups (SHAM, I/R , HBO-I/R and CONTROL). The surgical technique consisted of total clamping of the hepatic pedicle for 15 min followed by twice repeated reperfusion for 5 min (unclamping). HBO was applied in a collective chamber (simultaneous exposure of 4 rats) directly pressurized with oxygen at 2 ATA for 60 min. Hepatic mitochondrial function was determined using samples of the median lobe obtained after exactly 5 min of reperfusion for the analysis of mitochondrial respiration based on the determination of states 3 and 4, the respiratory control ratio and the transition of mitochondrial permeability (mitochondrial swelling).Data were analyzed by the Mann-Whitney test and the level of significance was set at p < 0.05. RESULTS: There was a statistically significant difference (p < 0.05) in state 3 between the CONTROL and I/R and HBO-I/R groups, in state 4 between the CONTROL and I/R and HBO-I/R groups; in respiratory control ratio (RCR) between the CONTROL and I/R and HBO-I/R groups and between the CONTROL and Sham groups, and in mitochondrial swelling between the CONTROL and I/R and HBO-/R groups and between the Sham and I/R and HBO-I/R groups. CONCLUSION: In this process of periodic ischemia and reperfusion, hyperbaric pre-conditioning did not improve significantly hepatic mitochondrial function.