Limonin inhibits apoptosis, promotes autophagy and improves non-alcoholic fatty liver disease by promoting oxidative metabolism of fatty acids through PPARα / 药学学报

Non-alcoholic fatty liver disease (NAFLD) is a very common chronic liver disease in clinic, which can further develop into liver fibrosis, cirrhosis, eventually hepatocellular carcinoma and liver failure. Limonin is a natural triterpenoid compound containing furan rings. Previous studies have found that limonin has good anti-inflammatory, analgesic and liver protective functions. However, the mechanism of action of limonin on NAFLD has not been clarified. Based on the background, C57BL/6J male mice were fed with high fat diet (HFD) to establish NAFLD model (the experiment was approved by the Animal Ethics Committee of Hefei University of Technology, the approval number is HFUT20220429001), and limonin was added to the mice for administration by intragastric administration (i.g.). The results showed that HFD can induce typical NAFLD phenotypes, including impaired liver function, increased fat accumulation, and increased serum aspartate amino transferase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP) levels in mice. Mice were treated with limonin (50 and 100 mg·kg-1) for 10 weeks, and it was found that limonin could restore dyslipidemia and improve fat accumulation in liver cells of mice. In addition, we conducted in vitro experiments with human hepatoma cell line HepG2 cells, and found that limonin can promote the expression of oxidative metabolism and autophagy related genes and inhibit apoptosis in HepG2 cells. Mechanistically, limonin improves high-fat food-induced NAFLD by promoting the expression of oxidative metabolism genes transcriptional coactivator of peroxisome proliferator activating receptor γ (PPARγ) (PGC1α) and carnitine palmitoyl transferase 1 alpha (CPT1α) through peroxisome proliferator activates receptor alpha (PPARα). These results indicate that limonin can inhibit apoptosis, promote autophagy and improve NAFLD by promoting oxidative metabolism of fatty acids through PPARα.

Effects of hyperbaric oxygen therapy on human psychomotor performance: A review / 中西医结合学报

Psychomotor performance is the coordination of a sensory or ideational (cognitive) process and a motor activity. All sensorimotor processes involved in planning and execution of voluntary movements need oxygen supply and seem to be significantly disrupted in states of hypoxia. Hyperbaric oxygen therapy has become a widely used treatment in routine medicine and sport medicine due to its beneficial effects on different aspects of human physiology and performance. This paper presents state-of-the-art data on the effects of hyperbaric oxygen therapy on different aspects of human psychomotor function. The therapy's influence on musculoskeletal properties and motor abilities as well as the effects of hyperbaric oxygenation on cognitive, myocardial and pulmonary functions are presented. In this review the molecular and physiological processes related to human psychomotor performance in response to hyperbaric oxygen are discussed to contribute to this fast-growing field of research in integrative medicine. Please cite this article as: Olex-Zarychta D. Effects of hyperbaric oxygen therapy on human psychomotor performance: A review. J Integr Med. 2023; 21(5): 430-440.

Fish oil supplementation and physical exercise during the development period increase cardiac antioxidant capacity in Wistar rats

Abstract Aim: To investigate if treadmill exercise (Ex) associated with fish oil (FO) supplementation during lactation would influence the biochemical profile as well as the oxidative balance in the hearts of male juvenile rats. Methods: Fifteen days-old rats were submitted to a daily moderate Ex training (based on their maximal running capacity) and FO supplementation for 4 weeks. Forty-eight hours after the last exercise session, blood fasting glucose and lipid profile were assessed according to the manufacturer's recommendations, while the oxidative status of the hearts was evaluated via colorimetric and absorbance-based assays. Results: FO associated with Ex decreased triglycerides (TG-79.27 ± 5.75 to 60.24 ± 6.25 mg/dL) and very-low-density lipoprotein cholesterol levels (VLDL-15.85 ± 1.15 to 12.05 ± 1.25 mg/dL) when compared to sedentary animals. FO, alone, reduced atherogenic index (AI- 1.14 ± 0.03 vs. 1.01 ± 0.04 a.u) while increased high-density lipoprotein cholesterol (HDL-43.90 ± 2.50 vs. 59.43 ± 3.15 mg/dL) of sedentary animals. Additionally, both Ex (67.3 ± 13.5 nmol/mg prot) and FO supplementation (56.6 ± 5.5 nmol/mg prot) decreased the oxidative damage to lipids in non-trained animals (105.8 ± 10.8 nmol/mg prot). The interventions also protected the protein content from oxidative stress (Ex- 5.15 ± 0.46; FO- 4.5 ± 0.5; and vehicle sedentary-7.3 ± 0.6 µmol/mg prot), while increasing the antioxidant defense and oxidative metabolism. Conclusion: Our findings suggest that intervention in juvenile rats can improve cardiac metabolism. These are the first findings to show the positive effects of the association between FO and moderate treadmill Ex during the critical period of development. We believe these results can drive early-life origins of heart disease through different avenues and, possibly, assist the development of a heart disease prevention program as well as an adjunctive therapeutic resource.

Simulation of the oxidative metabolization pattern of netupitant,an NK1 receptor antagonist,by electrochemistry coupled to mass spectrometry / 药物分析学报

Considering the frequent use of netupitant in polytherapy,the elucidation of its oxidative metabolization pattern is of major importance.However,there is a lack of published research on the redox behavior of this novel neurokinin-1 receptor antagonist.Therefore,this study was performed to simulate the intensive hepatic biotransformation of netupitant using an electrochemically driven method.Most of the known enzyme-mediated reactions occurring in the liver(i.e.,N-dealkylation,hydroxylation,and N-oxidation)were successfully mimicked by the electrolytic cell using a boron-doped diamond working electrode.The products were separated by reversed-phase high-performance liquid chromatography and identified by high-resolution mass spectrometry.Aside from its ability to pinpoint formerly unknown metabolites that could be responsible for the known side effects of netupitant or connected with any new perspective concerning future therapeutic indications,this electrochemical process also represents a facile alternative for the synthesis of oxidation products for further in vitro and in vivo studies.

Avaliação da atividade funcional dos neutrófilos sanguíneos em ovelhas primíparas e pluríparas da raça Lacaune / [Functional evaluation of blood neutrophils in primiparous and pluriparous Lacaune ewes]

This study aimed to evaluate the neutrophil oxidative metabolism and phagocytosis of Zymonsan particles of primiparous and pluriparous Lacaune ewes during the first 30 days after lambing. A total of 20 ewes were evaluated, 10 primiparous (GPR) and 10 pluriparous (GPL). Evaluation of basal oxidative metabolism was performed using the nitroblue tetrazolium (NBT) technique, stimulating neutrophil phagocytosis with Zymosan particles. Blood samples were collected at parturition day (M1) and 1, 3, 7, 15 and 30 days after parturition, corresponding to M2, M3, M4, M5 and M6, respectively. In relation to the groups, GPR presented lower oxidative basal metabolism neutrophils in M1 compared to M4, in M3 with M1, M2, M4 and M5. In M4 and M5 differences were found at all times and in M6 with M4 and M5. Higher percentage of neutrophils than phagocytes were found in M4, M5 and M6 than in M1 and M2 in GPL animals. At all times GPR presented a lower percentage of phagocytosis than GPL. Thus it is concluded that the immune response in pluriparous sheep was more efficient than in primiparous sheep.(AU)

Research progress on pyruvate dehydrogenase kinase inhibitors / 药学学报

Tumor cells can metabolize glucose through glycolysis to intermediates for biomacromolecule synthesis by inhibiting the activity of the pyruvate dehydrogenase complex (PDC) in mitochondria. In this process, pyruvate dehydrogenase kinases (PDKs) play a key role. The inhibition of the activity of PDKs can effectively block this metabolic pathway, activate mitochondrial oxidative metabolism, and induce tumor cell apoptosis. PDK inhibitors have become a research hotspot in medicinal chemistry, and novel structures targeting classical binding sites have been synthesized. In this paper, recent research progress on PDK inhibitors is reviewed to provide information on these latest entities and to explore their clinical applicability.

Estresse oxidativo em cães com doença periodontal: comparação dos biomarcadores plasmáticos e salivares / Oxidative stress in dogs with periodontal disease: comparison of plasmatic and salivary biomarkers

A doença periodontal (DP) é a enfermidade inflamatória mais comum da cavidade oral dos cães. A quantificação de biomarcadores do plasma e da saliva tem sido utilizada para avaliar o estresse oxidativo sistêmico (EOS) e local (EOL) da DP humana. Na DP canina, os mecanismos do estresse oxidativo não estão bem caracterizados e estabelecidos. O objetivo do presente estudo foi investigar a hipótese de que o EOS ocorre na DP canina e de que a saliva pode ser utilizada para avaliar o EOL. Analisou-se, também, a hipótese de que a ativação do metabolismo oxidativo dos neutrófilos contribui para EOS na DP dos cães. Para tal, foram selecionados 20 cães adultos portadores de DP, agrupados de acordo com o grau de lesão: gengivite (n=6), periodontites leve (n=8) e avançada (n=6). O grupo controle foi composto pelos mesmos 20 cães, 30 dias após o tratamento periodontal. Para avaliar o metabolismo oxidativo dos neutrófilos circulantes foi quantificada a produção de superóxido pelo teste de redução do tetrazólio nitroazul (NBT). As concentrações de oxidante total (TOC) e de espécies reativas ao ácido tiobartbitúrico (TBARS) no plasma foram quantificadas para avaliar o EOS. Para a avaliação do estresse oxidativo local, foi quantificado o TOC salivar e a concentração dos principais antioxidantes da saliva (albumina, ácido úrico e bilirrubina total). O EOS na DP foi confirmado pelo aumento da produção de superóxido dos neutrófilos circulantes, TOC e TBARS plasmático. Foi possível quantificar todos os biomarcadores na saliva de cães, porém nenhum foi capaz de expressar o EOL da DP canina. Esta é uma das primeiras evidências de que o EOS ocorre em cães com DP e que a ativação do metabolismo oxidativo dos neutrófilos pode contribuir para desequilíbrio entre antioxidantes e oxidantes. Este estudo ressalta a importância da higiene bucal dos cães para a prevenção da DP e de lesões degenerativas crônicas de diversos tecidos causadas pelo EOS.(AU)

Periodontal disease (PD) is the most common inflammatory disease of the oral cavity of dogs. Quantitation of plasma and salivary biomarkers have been used to assess the systemic oxidative stress (SOS) and local (LOS) of human PD. In canine PD, oxidative stress mechanisms are not well characterized and established. Our objective was to investigate the hypothesis that SOS occurs in dog PD and saliva can be used to evaluate the LOS. We also investigated the hypothesis that the activation of neutrophil oxidative metabolism contributes to SOS in dog SD. For this purpose, 20 adult dogs were selected PD patients, grouped according to the degree of injury: gingivitis (n=6), light periodontitis (n=8) and advanced periodontitis (n=6). The control group was composed of the same 20 dogs, 30 days after periodontal treatment. To assess oxidative metabolism of circulating neutrophils superoxide production was measured by test nitroblue tetrazolium reduction (NBT). The total oxidant concentrations (TOC) and reactive species to tiobartbitúrico acid (TBARS) in plasma were quantified to evaluate SOS. For the evaluation of local oxidative stress were quantified salivary TOC and concentration of the main antioxidant in saliva (albumin, uric acid, and total bilirubin). EOS in dogs with PD was confirmed by increased superoxide production of circulating neutrophils, TOC, and plasma TBARS. It was possible to quantify all the biomarkers in the saliva of dogs, but none was able to express the LOS canine PD. This is the first evidence that SOS occurs in dogs with PD and that activation of the oxidative metabolism of neutrophils may contribute to an imbalance between oxidants and antioxidants. This study highlights the importance of oral hygiene of dogs to prevent PD and chronic degenerative lesions of various tissues caused by SOS.(AU)

Research advances in non-P450-mediated drug oxidative metabolism / 药学学报

The major non-P450 enzymes involved in the oxidative metabolism of drugs are:the flavincontaining monooxygenase (FMO), the monoamine oxidase (MAO), the aldehyde oxidase (AO), the xanthine oxidase (XO), the alcohol dehydrogenase (ADH) and the aldehyde dehydrogenase (ALDH). In recent years, the role of non-P450 enzymes in drug oxidative metabolism has garnered increasing attention. However, the contribution of non-P450 enzymes to the drug oxidative metabolism is possibly underestimated in many cases, as most metabolism studies in drug discovery and lead optimization are conducted using in vitro test systems related to P450 enzymes. In this article, these non-P450 enzymes in terms of catalyzed reaction types, common substrates, gene polymorphism and drug interaction are reviewed, and the in vitro models and factors for non-P450-mediated oxidative metabolism are summarized. Similar to P450 enzymes, non-P450 enzymes can directly catalyze the oxidation of drugs, yielding therapeutically active metabolites or toxic metabolites. These enzymes can also oxidize the toxic metabolites, generated from P450-catalyzed reaction, to nontoxic metabolites. In general, most non-P450 enzymes (such as FMO and MAO) appear to be much less inducible than P450 enzymes.

Alteration of metabolic characteristics on the masseter muscle fiber of unilateral chewing rats and its adenosine monophosphate activated protein kinase regulatory mechanism / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aims to determine the influence of unilateral chewing on metabolic characteristics of masseter muscle fibers in rats and the regulatory effect of an adenosine monophosphate activated protein kinase (AMPK) signal pathway on metabolism.</p><p><b>METHODS</b>Rats were submitted to exodontia of all the right maxillary molars and divided into 2, 4, 6, and 8 weeks groups, and corresponding control groups were set as well. Sections were stained by nicotine adenine dinucleotide tetrazolim reductase(NADH-TRase) to demonstrate the types, proportion, and density of masseter muscle fibers. AMPKα1 and p-AMPK(Thr172) levels in bilateral masseter muscles were detected by Western blot.</p><p><b>RESULTS</b>In the 2-week group, the percentage of dark fibers augmented in the ipsilateral side, whereas the percentage of intermediary fibers in the contralateral side was increased accompanied by a decrease of light fibers, compared with the control group (P<0.05). The percentage of dark fibers was increased in the bilateral sides, whereas the percentage of dark fiber in the ipsilateral sides surpassed that of the contralateral sides in the 4, 6, and 8-week groups. The percentage of intermediary fibers was decreased in the bilateral sides in the 6 and 8-week groups (P<0.05). The percentage of light fibers was reduced in the ipsilateral sides in the 8-week group, whereas no alteration was observed in contralateral sides (P>0.05). In the ipsilateral sides, p-AMPK (Thr172)/AMPKα1 levels were increased in the 2 and 4-week groups (P<0.05), whereas no change was observed in the contralateral sides in either group (P>0.05).</p><p><b>CONCLUSIONS</b>Unilateral chewing increases the oxidative metabolic ability in bilateral masseter muscle fibers especially in the non-working side accompanied with change of muscle fiber types. The improvement of aerobic metabolism ability is related to the AMPK signal pathway.
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Comparação entre duas metodologias para a realização do teste de redução do tetrazólio nitroazul (NBT) em caprinos / Comparison among two methodologys at realization nitroblue tetrazolium reduction test (NBT) in caprines

O teste de redução do tetrazólio nitroazul (NBT) é um dos métodos mais utilizados para a avaliação do metabolismo oxidativo dos neutrófilos. Porém, o custo por amostra e o tempo de utilização do corante são desvantagens do método. O objetivo deste trabalho foi comparar duas metodologias para realização do teste do NBT a fim de maximizar a sua utilização e diminuir o custo por amostra processada. Foram utilizadas 10 cabras, adultas clinicamente sadias e colhidas amostras de sangue por meio de venipunção jugular em tubos de 10mL sendo retirada uma alíquota de 500µL em tubo "eppendorff" contendo 2µL de heparina. Através desta alíquota foram realizadas três metodologias para o teste do NBT: A) técnica original Park & Good (1970); B) redução de 50% do volume do corante; C) redução de 50% e armazenamento do corante a -20°C por 180 dias. Foram confeccionados e corados esfregaços sanguíneos (May Grünwald-Giemsa) das amostras com as metodologias propostas e realizada contagem de 100 neutrófilos em microscopia óptica para determinar o percentual de neutrófilos reativos ao NBT. A análise estatística pelo método de Análise de Medidas Repetida (ANOVA) demonstrou diferenças significativas (P<0,05) entre os dois métodos e o original (A=14,4±4,6; B=1,9±1,4; C=1,1±0,9), porém apresentou alta correlação pelo teste de Spearman entre os métodos (rs=0,82 (AxB), rs=0,75 (BxC) e rs=0,93 (AxC)). Conclui-se que o teste de Redução do Tetrazólio Nitroazul (NBT) segundo a metodologia descrita por Park & Good (1970) e a redução do corante em 50% do seu volume e a sua redução associada ao congelamento por 180 dias a -20ºC podem ser realizadas em caprinos, porém os valores devem ser comparados com as respectivas metodologias utilizadas.(AU)

The nitroblue tetrazolium reduction test (NBT) is one of the most used methods to evaluate the oxidative metabolism of neutrophils. However, the cost for each sample and dye's life-time are disadvantages of the method. This paper aim was to compare two NBT test methodologies in order to maximize its use and decrease the cost for each sample. It was made using 10 adult and healthy goats and blood samples were taken through venipuncion in jugular vein, using 10mL tubes, and then a 500µL sample was taken in eppendoff tubes with 2µL of heparin. Using this sample were made three methodologies for the NBT test: A) Park & Good (1970) original technique; B) dye reduction in 50% of the volume; C) 50% reduction and dye storage at -20°C for 180 days. It was made and dyed the blood smear (May Grünwald-Giemsa) from the samples, using the proposed methodologies and then made the counting of 100 neutrophils in optic microscopy, to determinate the percentage of NBT reactive neutrophils. The statistic analysis by the method of Repeated Measuring Analysis (ANOVA) evinced significant differences (P<0,05) between the two methods and the original (A=14.4±4.6; B=1.9±1.4; C=1.1±0.9), however it showed high correlation by the Spearman test between the methods (rs=0,82 (AxB), rs=0,75 (BxC) e rs=0.93 (AxC)). It was concluded that the original Park & Good (1970) method and two methodologies for dye reduction in 50% and it's reduction associated with NBT dye freezing for 180 days can be used in caprines, but the values should be compared with their respective methodologies.(AU)

Comparação entre duas metodologias para a realização do teste de redução do tetrazólio nitroazul (NBT) em caprinos

Abstract: The nitroblue tetrazolium reduction test (NBT) is one of the most used methods to evaluate the oxidative metabolism of neutrophils. However, the cost for each sample and dye's life-time are disadvantages of the method. This paper aim was to compare two NBT test methodologies in order to maximize its use and decrease the cost for each sample. It was made using 10 adult and healthy goats and blood samples were taken through venipuncion in jugular vein, using 10mL tubes, and then a 500L sample was taken in eppendoff tubes with 2L of heparin. Using this sample were made three methodologies for the NBT test: A) Park & Good (1970) original technique; B) dye reduction in 50% of the volume; C) 50% reduction and dye storage at -20°C for 180 days. It was made and dyed the blood smear (May Grünwald-Giemsa) from the samples, using the proposed methodologies and then made the counting of 100 neutrophils in optic microscopy, to determinate the percentage of NBT reactive neutrophils. The statistic analysis by the method of Repeated Measuring Analysis (ANOVA) evinced significant differences (P 0,05) between the two methods and the original (A=14.4±4.6; B=1.9±1.4; C=1.1±0.9), however it showed high correlation by the Spearman test between the methods (rs=0,82 (AxB), rs=0,75 (BxC) e rs=0.93 (AxC)). It was concluded that the original Park & Good (1970) method and two methodologies for dye reduction in 50% and it's reduction associated with NBT dye freezing for 180 days can be used in caprines, but the values should be compared with their respective methodologies.

Resumo: O teste de redução do tetrazólio nitroazul (NBT) é um dos métodos mais utilizados para a avaliação do metabolismo oxidativo dos neutrófilos. Porém, o custo por amostra e o tempo de utilização do corante são desvantagens do método. O objetivo deste trabalho foi comparar duas metodologias para realização do teste do NBT a fim de maximizar a sua utilização e diminuir o custo por amostra processada. Foram utilizadas 10 cabras, adultas clinicamente sadias e colhidas amostras de sangue por meio de venipunção jugular em tubos de 10mL sendo retirada uma alíquota de 500L em tubo "eppendorff" contendo 2L de heparina. Através desta alíquota foram realizadas três metodologias para o teste do NBT: A) técnica original Park & Good (1970); B) redução de 50% do volume do corante; C) redução de 50% e armazenamento do corante a -20°C por 180 dias. Foram confeccionados e corados esfregaços sanguíneos (May Grünwald-Giemsa) das amostras com as metodologias propostas e realizada contagem de 100 neutrófilos em microscopia óptica para determinar o percentual de neutrófilos reativos ao NBT. A análise estatística pelo método de Análise de Medidas Repetida (ANOVA) demonstrou diferenças significativas (P 0,05) entre os dois métodos e o original (A=14,4±4,6; B=1,9±1,4; C=1,1±0,9), porém apresentou alta correlação pelo teste de Spearman entre os métodos (rs=0,82 (AxB), rs=0,75 (BxC) e rs=0,93 (AxC)). Conclui-se que o teste de Redução do Tetrazólio Nitroazul (NBT) segundo a metodologia descrita por Park & Good (1970) e a redução do corante em 50% do seu volume e a sua redução associada ao congelamento por 180 dias a -20ºC podem ser realizadas em caprinos, porém os valores devem ser comparados com as respectivas metodologias utilizadas.

Protection Effect of Wogonin on Mice with Hyperlipidemia / 医药导报

Objective To explore the effects of wogonin on hyperlipidemia in mice and clarify the molecule mechanism. Methods Thirty mice were evenly divided into three group normal control group,model control group and treatment group. The normal control group was given normal diet,the model control group received high-fat diet,the treatment group received high-fat diet with wogonin (500 mg·kg-1 ). Results The mice developed hyperlipidemia 12 weeks after starting the high-fat diet. The body weight,visceral fat and fat index were increased (P<0. 05). After treatment,these indices were reduced ( P < 0. 01). Wogonin significantly reduced the total cholesterol ( TC),low density lipoprotein ( LDL),high density lipoprotein (HDL),except the triglyceride (TG). Compared to the model control group,the hepatic lipase(HL) and lipoprotein lipase(LPL) activity in the treatment group were recovered (P<0. 05),but HMG-CoA reductase activity was inhibited ( P<0. 01). Mechanistic study suggested that the lipid-lowering effect might be related to the lipid synthesis genes (SREBP-1c,FAS, PPARγ) and the lipid metabolism genes (PPARα,CPT-1). Conclusion Wogonin can prevent hyperlipidemia,which might be related to the regulation of enzyme activity and the changes of lipid synthesis and oxidative metabolism.

Oxidative metabolism and muscle biochemical profile of polo horses supplemented with an ADE vitamin complex / Metabolismo oxidativo e perfil bioquímico muscular de cavalos de pólo suplementados com complexo vitamínico ADE

Horses used for the game of polo experience abrupt and frequent changes in exercise intensity. To meet this variable energy demand, the horses use both aerobic and anaerobic pathways in varying proportions and intensities. In this context, there must be a balance between the formation of reactive oxygen species (ROS) and the action of antioxidants to prevent oxidative stress and its consequences. The effect of supplementation with an ADE vitamin complex on oxidative metabolism was evaluated in 18 crossbred horses randomly divided between a treated group (TG) and a control group (CG). The TG animals received the ADE vitamin complex (1mL/50 kg of body weight) by deep intramuscular injection at 30 and 15 days before the game. The CG horses received 10ml of saline by the same administration route and schedule. During the polo match, the animals played for a total of 7.5 min. Blood samples were collected on the same days as the treatments were administered, and immediately before and at 15, 90 and 180 minutes after the game. The concentrations of creatine phosphokinase (CK), lactate dehydrogenase (LDH), lactate, glucose, aspartate aminotransferase (AST), glutathione (GSH), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured in the blood samples. After the game, the TG demonstrated higher levels of AST, lactate and glucose than the CG, suggesting more efficient energy use by the treated animals. The higher GSH and lower lactate levels in the TG before the game suggest the presence of a greater antioxidant supply in the treated animals. The maintenance of the MDA levels indicates that neither of the groups exhibited oxidative stress.

O jogo de pólo se caracteriza por mudanças abruptas e frequentes na intensidade do exercício dos cavalos. Para satisfazer esta demanda inconstante de energia, os animais utilizam as vias aeróbia e anaeróbia em proporções e intensidade variáveis. Neste contexto deve haver equilíbrio entre a formação das espécies reativas de oxigênio (EROs) e a ação das substâncias antioxidantes a fim de evitar o estresse oxidativo e suas consequências. Avaliou-se o efeito da suplementação com vitaminas ADE no metabolismo oxidativo destes animais. Para tanto, 18 equinos mestiços foram distribuídos aleatoriamente em dois grupos: tratado e controle (GT) e controle (GC).Os animais do GT receberam complexo vitamínico ADE (1 mL/50 kg de peso vivo) pela via intramuscular profunda aos 30 e 15 dias antes do jogo. Os cavalos do GC receberam, pela mesma via de administração e nos mesmos momentos, 10mL de solução fisiológica. Os animais jogaram um tempo de 7,5min. Foram coletadas amostras de sangue nos mesmos dias de tratamento e imediatamente antes e aos 15, 90 e 180 minutos após o jogo. Foram determinadas as concentrações sanguíneas de CK, LDH, lactato, glicose, AST, GSH, SOD e MDA. Após o jogo o GT apresentou maiores valores para AST, lactato e glicemia que o GC, sugerindo melhor aproveitamento energético dos animais tratados. Os valores maiores de GSH e menores de lactato no GT antes da prova sugerem maior aporte antioxidante nos animais tratados. A manutenção dos teores de MDA indica que nenhum dos grupos entrou em estresse oxidativo.

Estresse oxidativo e aumento da apoptose em neutrófilos de cães com azotemia pré-renal / Oxidative stress and increase in apoptosis index of neutrophils from dogs with prerenal azotemia

O presente trabalho tem como objetivo testar a hipótese de que, à semelhança do que ocorre na uremia, cães com azotemia pré-renal sofrem estresse oxidativo, o qual está relacionado com alterações do metabolismo oxidativo e apoptose dos neutrófilos. Para tal, foi determinada a peroxidação lipídica pela quantificação do malondialdeído (MDA) e o status antioxidante total do plasma de 15 cães normais e 10 com azotemia pré-renal, correlacionando-os com a produção de superóxido e o índice apoptótico dos neutrófilos. As determinações do MDA e do status antioxidante total foram estabelecidas empregando-se um conjunto de reagentes comerciais. Por meio de citometria de fluxo capilar, a produção de superóxido e a apoptose de neutrófilos isolados de sangue periférico foram determinadas utilizando-se a sonda hidroetidina e o sistema anexina V-PE, respectivamente. Cães azotêmicos (26,29±5,32g/L) apresentaram menor concentração (p=0,0264) do antioxidante albumina em relação ao grupo-controle (30,36±3,29g/L) e também uma menor (p=0,0027) capacidade antioxidante total (2,36±0,32 versus 2,73±0,24mmol/L), enquanto não houve alteração da peroxidação lipídica plasmática e da produção de superóxido neutrofílica. Concluiu-se que, à semelhança do que ocorre na uremia, condições azotêmicas pré-renais no cão causam estresse oxidativo e aceleração da apoptose dos neutrófilos.

This study aims to test the hypothesis that, similarly to what occurs in uremia, dogs with prerenal azotemia suffer oxidative stress associated with changes in oxidative metabolism and apoptosis in neutrophils. For this purpose, fifteen normal dogs and ten with prerenal azotemia had lipid peroxidation determined by quantifying the malondialdehyde (MDA) and had plasma total antioxidant status evaluated, correlating them with the superoxide production and apoptotic index of neutrophils. MDA and plasma total antioxidant status were determined using commercial reagents. Using capillary flow cytometry, superoxide production and apoptosis were determined from isolated neutrophils of peripheral blood using the hydrithidine and Annexin V-PE probe system, respectively. Azotemic dogs (26.29±5.32g/L) had a lower concentration (p=0.0264) of the plasma antioxidant albumin than the control group (30.36±3.29g/L) and also had lower (p=0.0027) total antioxidant status (2.36±0.32 versus 2.73±0.24mmol/L), while no alterations were observed in plasma lipid peroxidation and superoxide production. It was concluded that, similarly to what occurs in uremia, prerenal azotemia causes oxidative stress and acceleration of neutrophil apoptosis in dogs.

Peripheral skeletal muscle dysfunction in rats with emphysema / 中华物理医学与康复杂志

Ohjective To study the effects of emphysema on peripheral skeletal muscle biomechanics,pathomorphology and oxidative metabolism in rats.Methods Twenty Sprague-Dawley rats were used and randomized equally into a control group and an emphysema group.A dose of 40 U/100 g body weight of porcine pancreatic elastase was instilled into the trachea of the animals of the emphysema group to model emphysema,while the same volume of saline was instilled into those of the control group.Twenty weeks after instillation,in situ mechanical properties of gastrocnemius were evaluated.Gastrocnemius fiber type composition and capillary density (CD) were assessed by using ATPase staining.Lipofuscin accumulation (LI/F) was determined with the ferric-ferricyanide reduction test technique.Immunohistochemistry was used for the detection of nitric oxide synthases (NOS) in gastrocnemius.The muscle biopsy homogenate was used to measure the activity of superoxide dismutase (SOD),catalase (CAT),NOS,total antioxidant capacity (T-AOC) and the content of nitric oxide (NO).Results Emphysema increased fatigability and decreased the recovery rate of gastrocnemius muscle [(145.0 ± 55.4) s vs (55.2 ± 29.3)s,P ＜ 0.05].Compared to control,the gastrocnemius muscle in rats with emphysema had a lower CD [(513.9±71.1)n/mm2 vs (578.6 ±59.9)n/mm2,P ＜0.05] and a decreased proportion of type I fibers [(16.0 ±5.0)％ vs (30.7 ±4.1) ％,P ＜0.05],with a reciprocal increase in type II b/x fibers [(27.3 ±4.8)％vs (11.0±3.2)％,P＜0.05].LI/F was higher (3.3±0.5 vsl.7±0.4,P＜0.05) and the activity of SOD,CAT and T-AOC was increased in emphysema group.Compared with control,rats with emphysema demonstrated a lower expression of endothelial NOS (eNOS) (1.9 ± 0.5 vs 3.4 ± 0.6,P ＜ 0.05),and an equivalent expression of neuronal NOS (nNOS) (4.7 ± 1.0 vs 5.1 ± 0.8,P ＞ 0.05) in the gastrocnemius muscle.The inducible NOS (iNOS) was not found in both groups.Conclusions Emphysema could induce biomechanical,pathomorphological and oxidative metabolic changes in peripheral skeletal muscle.

Efeito do tempo de conservação de polimorfonucleares do sangue de bezerros sobre o metabolismo oxidativo e a atividade de fagocitose de Escherichia coli / Effect of conservation time on bovine polymorphonuclear leukocytes in calves blood on oxidative burst and Escherichia coli phagocytosis activity

Foram avaliados os efeitos do tempo sobre o metabolismo oxidativo e a fagocitose de Escherichia coli, em amostras de polimorfonucleares (PMN) do sangue, de cinco bezerros hígidos, conservadas em banho de gelo por duas (t2), quatro (t4), seis (t6), 12 (t12) e 24 (t24) horas. O metabolismo oxidativo foi avaliado utilizando o Diacetato 2' 7' Diclorofluoresceína (DCFH-DA) e a E. coli, como estímulo. Para a fagocitose a mesma bactéria foi utilizada. As amostras foram analisadas por citometria de fluxo. O metabolismo oxidativo basal dos PMN do sangue de bezerros foi maior nos tempos t4, t6 e t12, do que em t2 (p<0,05). A intensidade de fluorescência do metabolismo oxidativo induzido pela bactéria foi maior nos tempos t4 e t6 do que em t2 (p<0,05). A comparação entre o metabolismo basal e induzido pela bactéria, em cada um dos tempos, mostrou que a maior diferença ocorreu em t2, com valores da média geométrica e desvios padrão respectivos de 18,3 ± 4,4 e 26,7 ± 1,8 (p< 0,05). A atividade de fagocitose, medida pela intensidade de fluorescência, foi maior para as amostras mantidas em gelo por 6 horas do que para t2, t4 e t12 (p<0,05). O percentual de fagocitose não diferiu entre os tempos. O tempo ideal para análise do metabolismo oxidativo foi o deduas horas. Maiores estudos são necessários para se verificar a influência do tempo de conservação na fagocitose de E. coli por PMN do sangue de bovinos.

The time effect on oxidative burst and phagocytosis of Escherichia coli by polymorphonuclear leukocytes of 5 healthy calve blood samples were evaluated. Samples were kept on ice bath for two (t2), four (t4), six (t6), twelve (t12) and 24 hours (t24) before the tests. The oxidative burst was measured using the 2',7'-dichlorfluorescein-diacetate (DCFH-DA) and E.coli as stimulus, also the same bacteria was used to phagocytosis process. The PMN basal oxidative burst of calves blood samples were analyzed by flow cytometry and were higher at times t4, t6 and t12 (p<0,05) than at t2. The oxidative burst fluorescence intensity induced by bacteria was higher at the times t4 and t6 than at t2 (p<0,05). The comparison between the basal oxidativeburst and the one induced by bacteria on each time, showed t2 time as the one with the highest difference, with geometric media and standard deviation of 18,3 ± 4,4 and 26,7 ± 1,8 (p< 0,05), respectively. Phagocytosis activity was measured by fluorescence intensity and was higher on samples kept on ice bath for 6 hours than those at t2, t4 and t12 (p<0,05). The phagocytosis percentage did not differ between the different times. The ideal time to test the oxidative burst was 2 hours (t2). Further researches are necessary to verify the conservation time influence on E.coli phagocytosis by PMN of bovine blood.

Effect of Chronical Ethanol Ingestion on the Levels of Fatty Acid Ethyl Esters (FAEEs) and Lipid Peroxidation in Rat Tissues

The present study was designed to observe the effect of chronically ingested ethanol on the level of fatty acid ethyl esters (FAEEs), which is a non-oxidative metabolite of ethanol metabolism in tissues, and its correlation to the status of oxidative stress in rats. Forty male Sprague Dawley rats weighing 145 - 155 g were divided into 2 groups, Control and EtOH. All rats were fed Lieber-DeCarli liquid diet for 4 weeks by pair-feeding. An isocaloric maltose dextrin was added in replace of 50 g ethanol (36%kcal) in the control diet. Chronically ingested ethanol significantly increased the content of FAEEs in pancreas and liver, but not in brain. The level of 2-thiobarbituric acid reactive substances (TBARS) was significantly increased, but alpha-tocopherol level was significantly decreased in pancreas and liver. However, the levels of TBARS and alpha-tocopherol in brain were not significantly affected by ethanol ingestion. Therefore, chronically ingested ethanol might cause tissue damage by increasing the levels of FAEEs and TBARS and dissipating more alpha-tocopherol in tissues.

Enzyme histochemical study of germanium dioxide-induced mitochondrial myopathy in rats

The purpose of this study were 1) to determine the earliest pathological changes of germanium dioxide (GeO2)-induced myopathy; 2) to determine the pathomechanism of GeO2-induced myopathy; and 3) to determine the minimal dose of GeO2 to induce myopathy in rats. One hundred and twenty five male and female Sprague-Dawley rats, each weighing about 150 gm, were divided into seven groups according to daily doses of GeO2. Within each group, histopathological studies were done at 4, 8, 16, and 24 weeks of GeO2 administration. Characteristic mitochondrial myopathy was induced in the groups treated daily with 10 mg/kg of GeO2 or more. In conclusion, the results were as follows: 1) The earliest pathological change on electron microscope was the abnormalities of mitochondrial shape, size and increased number of mitochondria; 2) The earliest pathological change on light microscope was the presence of ragged red fibers which showed enhanced subsarcolemmal succinate dehydrogenase and cytochrome c oxidase reactivity; 3) GeO2 seemed to affect the mitochondrial oxidative metabolism of muscle fibers; 4) GeO2 could induce mitochondrial myopathy with 10 mg/kg of GeO2 for 4 weeks or less duration in rats.

Evaluation of the Influence of Thermal Injury on Bactericidal Capacity of Neutrophils through Studies on Changes of Cell Oxidative Metabolism / 第三军医大学学报

Body defence functions are seriously impaired after thermal injury.Neutrophils,the important phagocytic cells against bacterial invasion,are also affected Therefore it is of value to study the functional statue of these cells in the postburn course.Two assays to observe the bactericidal function of neutrophils were adopted: the determination of the amount of H2O2 released and the126I taken by the activated neutrophils in oxidative metabolic processes.The results of the experiments on 2 animal models,inhalation injured dogs and scalded rats (TBSA 30% of third degree burns) with pseudomonas infection,showed that the cell oxidative metabolic response of the neutrophils was severely depressed in the first week postburn,indicating the presence of profound inhibition of the bactericidal function of the cells after thermal injury.In order to verify these results,direct yeast-killing assay was carried out and similar results were obtained.In order to explore the cause of the suppression of the bactericidal function,whether it was due to the presence of suppressive factors in the blood of burn victims,or due to the direct detrimental effect of thermal injury on the cells,further experiments to observe the crossed effects of burned serum on normal cells and normal serum on burned cells were performed.It was demonstrated that serum suppressive factors were present and the direct effects from thermal injury also played a part in suppressing the function