RESUMO
There are currently no available FDA-cleared biodosimetry tools for rapid and accurate assessment of absorbed radiation dose following a radiation/nuclear incident. Previously we developed a protein biomarker-based FAST-DOSE bioassay system for biodosimetry. The aim of this study was to integrate an ELISA platform with two high-performing FAST-DOSE biomarkers, BAX and DDB2, and to construct machine learning models that employ a multiparametric biomarker strategy for enhancing the accuracy of exposure classification and radiation dose prediction. The bioassay showed 97.92% and 96% accuracy in classifying samples in human and non-human primate (NHP) blood samples exposed ex vivo to 0-5 Gy X-rays, respectively up to 48 h after exposure, and an adequate correlation between reconstructed and actual dose in the human samples (R2 = 0.79, RMSE = 0.80 Gy, and MAE = 0.63 Gy) and NHP (R2 = 0.80, RMSE = 0.78 Gy, and MAE = 0.61 Gy). Biomarker measurements in vivo from four NHPs exposed to a single 2.5 Gy total body dose showed a persistent upregulation in blood samples collected on days 2 and 5 after irradiation. The data indicates that using a combined approach of targeted proteins can increase bioassay sensitivity and provide a more accurate dose prediction.
Assuntos
Biomarcadores , Proteínas de Ligação a DNA , Proteína X Associada a bcl-2 , Animais , Humanos , Biomarcadores/sangue , Proteínas de Ligação a DNA/sangue , Proteína X Associada a bcl-2/metabolismo , Proteína X Associada a bcl-2/sangue , Exposição à Radiação/efeitos adversos , Masculino , Radiometria/métodos , Macaca mulatta , Feminino , Aprendizado de Máquina , Doses de RadiaçãoRESUMO
Honeybee products consist of many substances, which have long been known for their medicinal and health-promoting properties. This study set out to appraise the protective potential of Egyptian propolis (EP) and bee venom (BV) separately or combined against total body irradiation (TBI) induced oxidative injury in rats. Besides, we assessed the bioactive components in EP and BV using HPLC and UPLC/ ESI-MS analysis in the positive ion mode. The animals were subjected to a source of gamma ionizing radiation at a dose of 6 Gy. Propolis and BV were administered independently and in combination before 14 days of γ-irradiation. Liver and kidney functions were estimated besides, DNA damage index (8- OHdG) by ELISA. Antioxidants, including glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were detected. Gene expression technique investigated for BAX, BCL2, and in plasma also miR125b expression in serum of rats. Besides, the histopathological for the brain, liver, kidney, and heart were investigated. In addition, lipid peroxidation was investigated in plasma and in the previous organs. The present results provide opportunities to advance the use of bee products as promising medicinal sources.
Assuntos
Venenos de Abelha/farmacologia , Raios gama/efeitos adversos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Própole/farmacologia , Protetores contra Radiação , Animais , Antioxidantes/metabolismo , Venenos de Abelha/administração & dosagem , Venenos de Abelha/química , Dano ao DNA/efeitos dos fármacos , Técnicas In Vitro , Própole/administração & dosagem , Própole/química , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Ratos , Proteína Supressora de Tumor p53/sangue , Proteína X Associada a bcl-2/sangueRESUMO
PURPOSE: Among abused substances, methamphetamine is a psychostimulant drug widely used recreationally with public health importance. This study investigated the effect of methamphetamine on proliferation, differentiation, and apoptosis of human adipose tissue stem cells (AdSCs). METHODS: AdSCs were isolated from human abdominal adipose tissue and were characterized for mesenchymal properties and growth kinetics. MTT assay was undertaken to assess methamphetamine toxicity on proliferation and differentiation properties and apoptosis of hAdSCs. RESULTS: Isolated cells were shown to have mesenchymal properties and a population doubling time (PDT) of 40.1 h. Following methamphetamine treatment, expressions of KI-67 and TPX2 as proliferation genes and Col1A1 and PPARg as differentiation genes decreased. Methamphetamine administration increased the expression of Bax and decreased Bcl-2 genes responsible for apoptosis. CONCLUSIONS: Our data suggested when AdSCs were exposed to methamphetamine, it decreased proliferation and differentiation properties of stem cells together with an increase in apoptosis. These findings can be added to the literature, especially when methamphetamine is used recreationally for weight loss purposes.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Metanfetamina/farmacologia , Tecido Adiposo/citologia , Proteínas de Ciclo Celular/efeitos dos fármacos , Genes bcl-2/efeitos dos fármacos , Humanos , Antígeno Ki-67/efeitos dos fármacos , Proteínas Associadas aos Microtúbulos/efeitos dos fármacos , PPAR gama/efeitos dos fármacos , Proteína X Associada a bcl-2/sangue , Proteína X Associada a bcl-2/efeitos dos fármacosRESUMO
An inadequate platelet response to aspirin (ASA) has been identified in some patients under chronic ASA treatment. The aim of this study was to analyze if ASA-sensitive and ASA-resistant platelets have differences in their apoptotic capability. Clinically stable ischemic coronary patients who had been taking ASA (100 mg/d) for at least 9 months before inclusion were divided into ASA-resistant (n = 11) and ASA-sensitive (n = 13) groups as defined by the PFA-100 test. Platelets from ASA-sensitive patients showed higher expression of the proapoptotic proteins Bak and Bax than those from ASA-resistant patients, although only Bak protein remained different when the results were adjusted by age. In resting platelets, neither caspase-3 activity nor cytosolic cytochrome C levels were different between both experimental groups. Stimulation of platelets with calcium ionophore (10 nmol/L, A23187) increased caspase-3 activity (1.91-fold higher; P < 0.05) and cytosolic cytochrome C levels (1.84-fold higher; P < 0.05) to a higher degree in ASA-sensitive than in ASA-resistant platelets. In conclusion, ASA-sensitive platelets seem to be better prepared to undergo apoptosis during robust platelet activation.
Assuntos
Proteínas Reguladoras de Apoptose/sangue , Apoptose/efeitos dos fármacos , Aspirina/uso terapêutico , Plaquetas/efeitos dos fármacos , Isquemia Miocárdica/tratamento farmacológico , Inibidores da Agregação Plaquetária/uso terapêutico , Idoso , Plaquetas/metabolismo , Plaquetas/patologia , Calcimicina/farmacologia , Ionóforos de Cálcio/farmacologia , Caspase 3/sangue , Resistência a Medicamentos , Complexo IV da Cadeia de Transporte de Elétrons/sangue , Feminino , Humanos , Masculino , Isquemia Miocárdica/sangue , Isquemia Miocárdica/patologia , Ativação Plaquetária/efeitos dos fármacos , Resultado do Tratamento , Proteína Killer-Antagonista Homóloga a bcl-2/sangue , Proteína X Associada a bcl-2/sangueRESUMO
OBJECTIVE: Acute cerebral infarction (ACI) is the most common type of acute cerebrovascular disease so far, and its incidence rate has been increasing in recent years. At present, the methods of diagnosing ACI in clinic are extremely complicated, and an effective index that can effectively diagnose ACI is urgently needed in clinic. This study is designed to investigate the clinical significance of Follistatin-like protein 1 (FSTL1), Bax and Bcl-2 in ACI. PATIENTS AND METHODS: A total of 84 cases of ACI patients admitted to our hospital from September 2017 to September 2019 and 90 cases of healthy subjects undergoing physical examination at the same time were selected as the research objects for prospective analysis. The concentrations of FSTL1, Bax and Bcl-2 in the peripheral blood of objects in the two groups were detected to analyze the diagnostic value of FSTL1, Bax and Bcl-2 for ACI, and the correlation of FSTL 1, Bax and Bcl-2 with the infarct size, treatment method and hemorrhagic transformation. Another 20 SD rats were purchased, among which 10 rats were randomly selected for ACI modeling. FSTL1 concentration, Bax and Bcl-2 protein expression in brain tissues of ACI rats and normal rats were detected. RESULTS: FSTL1 and Bax in peripheral blood of ACI patients were higher than those of healthy subjects (p<0.050), and Bcl-2 was lower than those of healthy subjects (p<0.050). It was detected that FSTL1, Bax and Bcl-2 had good diagnostic value for patients with ACI (p<0.001). FSTL1 and Bax decreased while Bcl-2 increased in patients treated with thrombolytic therapy (p<0.050). And FSTL1, Bax and Bcl-2 were closely related to infarct size and hemorrhagic transformation (p<0.050). Logistic regression analysis showed that NIHSS score, atrial fibrillation, infarct volume, FSTL1 and Bax were independent risk factors affecting hemorrhagic transformation in ACI patients (p<0.050), and Bcl-2 was an independent protective factor affecting hemorrhagic transformation in ACI patients (p<0.050). The concentration of FSTL1 and the expression of Bax protein in rat brain tissue were also higher than that in normal rats, while Bcl-2 was lower than that in normal rats (p < 0.001). CONCLUSIONS: FSTL1, Bax and Bcl-2 are involved in the occurrence and development of ACI and are closely related to the hemorrhagic transformation of patients. The mechanism by which FSTL1 promotes the occurrence of ACI might be related to promoting the occurrence of inflammatory responses in the brain tissue of patients or accelerating the apoptosis of neurons.
Assuntos
Hemorragia Cerebral/tratamento farmacológico , Infarto Cerebral/tratamento farmacológico , Terapia Trombolítica , Doença Aguda , Animais , Hemorragia Cerebral/sangue , Infarto Cerebral/sangue , Proteínas Relacionadas à Folistatina/sangue , Humanos , Modelos Logísticos , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Ratos , Ratos Sprague-Dawley , Proteína X Associada a bcl-2/sangueRESUMO
BACKGROUND: Early single-dose treatment with human mesenchymal stem cell-derived exosomes promotes neuroprotection and promotes blood-brain barrier integrity in models of traumatic brain injury (TBI) and hemorrhagic shock (HS) in swine. The impact of an early single dose of exosomes on late survival (7 days), however, remains unknown. We sought to evaluate the impact of early single-dose exosome treatment on neurologic outcomes, brain lesion size, inflammatory cytokines, apoptotic markers, and mediators of neural plasticity in a 7-day survival model. METHODS: Yorkshire swine were subjected to a severe TBI (8-mm cortical impact) and HS (40% estimated total blood volume). After 1 hour of shock, animals were randomized (n = 4/cohort) to receive either lactated Ringer's (5 mL) or lactated Ringer's with exosomes (1 × 10 exosome particles). After an additional hour of shock, animals were resuscitated with normal saline. Daily neurologic severity scores were compared. At 7 days following injury, lesion size, inflammatory markers, and mediators of inflammation (NF-κB), apoptosis (BAX), and neural plasticity (brain-derived neurotrophic factor) in brain tissue were compared between groups. RESULTS: Exosome-treated animals had significantly lower neurologic severity scores (first 4 days; p < 0.05) and faster neurologic recovery. At 7 days, exosome-treated animals had significantly smaller (p < 0.05) brain lesion sizes. Exosome-treated animals also had significantly lower levels of inflammatory markers (interleukin [IL]-1, IL-6, IL-8, and IL-18) and higher granulocyte-macrophage colony-stimulating factor levels compared with the control animals, indicating specific impacts on various cytokines. The BAX and NF-κB levels were significantly lower (p < 0.05) in exosome-treated animals, while brain-derived neurotrophic factor levels were significantly higher (p < 0.05) in the exosome-treated animals. CONCLUSION: In a large animal model of TBI and HS, early single-dose exosome treatment attenuates neurologic injury, decreases brain lesion size, inhibits inflammation and apoptosis, and promotes neural plasticity over a 7-day period.
Assuntos
Lesões Encefálicas Traumáticas/fisiopatologia , Lesões Encefálicas Traumáticas/terapia , Exossomos , Neuroproteção , Choque Hemorrágico/fisiopatologia , Choque Hemorrágico/terapia , Animais , Apoptose , Barreira Hematoencefálica , Lesões Encefálicas Traumáticas/patologia , Citocinas/sangue , Modelos Animais de Doenças , Feminino , Hemodinâmica , Inflamação/patologia , Células-Tronco Mesenquimais/citologia , NF-kappa B/sangue , Choque Hemorrágico/patologia , Transdução de Sinais , Suínos , Resultado do Tratamento , Proteína X Associada a bcl-2/sangueRESUMO
BCL2 and BAX genes are a group of signalling inducer and inhibitor genes playing a key role in the process of cellular physiological death (apoptosis). These genes, through the JAK/STAT signalling pathway, affect different cytokines on cell function and subsequently lead to the pathophysiology of diseases, especially autoimmune diseases. In addition, altering the methylation of genes can affect their expression. Since the aetiology and pathology of Behcet's disease is not fully understood, the aim of this study was to determine the methylation pattern of BCL2 and BAX genes in patients with Behcet's disease and compare it with those of control group. This was a case-control study on 51 patients with Behcet and 61 control subjects. Blood samples were received from all subjects. Subsequently, the peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll method and the methylation of the sites was investigated using quantitative methylation specific PCR (qMS-PCR) technique after extraction of DNA by salting out method and its examination with Nano drop. The results of methylation and expression of Bax gene suggest that the methylation level in the patient group significantly increased compared to the healthy individuals (p-value < .05). Furthermore, the results related to Bax gene expression revealed that the mean of gene expression in the patient group has decreased compared to the healthy group, and this decrease was statistically significant (p-value < .05). The rate of expression and methylation of Bcl2 did not indicate any change in the two patient and healthy groups. Given the results of this study, it can be guessed that perhaps DNA methylation is involved in certain conditions of the disease and it may result in regulation of the expression of the involved genes such as Bax gene, in the pathogenesis of the disease.
Assuntos
Síndrome de Behçet/sangue , Metilação de DNA/genética , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteína X Associada a bcl-2/sangue , Adulto , Apoptose/genética , Síndrome de Behçet/genética , Síndrome de Behçet/patologia , Feminino , Regulação da Expressão Gênica/genética , Humanos , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteína X Associada a bcl-2/genéticaRESUMO
AIM: Disturbed placentation results in pregnancy complications like preeclampsia. Placental development is influenced by apoptosis during trophoblast differentiation and proliferation. Increased oxidative stress upregulates placental apoptosis. We have earlier reported increased oxidative stress, lower omega-3 fatty acids and vitamin E levels in women with preeclampsia. Current study examines effect of maternal omega-3 fatty acids and vitamin E supplementation on apoptotic markers across gestation in a rat model of preeclampsia. MAIN METHODS: Pregnant Wistar rats were randomly assigned to control; early onset preeclampsia (EOP); late onset preeclampsia (LOP); early onset preeclampsia + omega-3 fatty acid + vitamin E supplementation (EOP + O + E) and late onset preeclampsia + omega-3 fatty acid + vitamin E supplementation (LOP + O + E) groups. Animals (Control, EOP, EOP + O + E) were sacrificed at d14 and d20 of gestation while animals (LOP, LOP + O + E) were sacrificed at d20 to collect blood and placentae. Protein and mRNA levels of apoptotic markers were analyzed by ELISA and RT-PCR respectively. KEY FINDINGS: Protein levels of proapoptotic markers like Bcl-2 associated X-protein (BAX) (pâ¯<â¯0.05), caspase-8 and 3 (pâ¯<â¯0.01 for both) and malondialdehyde (pâ¯<â¯0.01) were higher only in the EOP group as compared to control. However, the antiapoptotic marker, B cell lymphoma 2 (Bcl-2) protein levels were lower in both the subtypes of preeclampsia (pâ¯<â¯0.01 for both). SIGNIFICANCE: Our findings suggest that supplementation was beneficial in reducing the caspase-8 and 3 in early onset preeclampsia but did not normalize BAX and Bcl-2 levels. This has implications for reducing placental apoptosis in preeclampsia.
Assuntos
Ácidos Graxos Ômega-3/uso terapêutico , Pré-Eclâmpsia/dietoterapia , Vitamina E/uso terapêutico , Animais , Apoptose/fisiologia , Biomarcadores/metabolismo , Caspase 3/análise , Caspase 3/sangue , Caspase 8/análise , Caspase 8/sangue , Suplementos Nutricionais , Modelos Animais de Doenças , Ácidos Graxos Ômega-3/metabolismo , Feminino , Masculino , Fenômenos Fisiológicos da Nutrição/fisiologia , Estresse Oxidativo/fisiologia , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Vitamina E/metabolismo , Proteína X Associada a bcl-2/análise , Proteína X Associada a bcl-2/sangueRESUMO
INTRODUCTION: Abnormal behavior can cause harm or loss to oneself, the family, and society and may be related to psychological endurance levels. With early identification and early intervention, the occurrence of harm can be prevented and the loss can be reduced. Now there is no clear definition of psychological endurance levels and no accurate measurement tools yet. METHODS: This study first proposes the concept of psychological endurance threshold (PET) and defines that as: "the psychological state threshold of human objective physiological characteristics and outbreaks of abnormal behavior led by subjective cognitive level difference". The study hypothesizes that human behavior is related to it, and constructs multiple measurement method tools to measure it. RESULTS: Here we show PET exists objectively and can be measured exactly by methods such as psychological endurance threshold measurement table, experience evaluation, dopamine level detection, and genetic testing. In particular, PET is determined by AKT1, PRDM4, and BAX which are the natural markers of PET. CONCLUSIONS: The significance of this study is to discover people with abnormal expression of AKT1, PRDM4, and BAX who have lower PET and tend to commit abnormal behavior more easily. Understanding PET will enable people to make self-adjustment or to intervene by professionals as soon as possible and in a timely manner in the face of various negative stimuli in work and life, especially for people with lower PET, people should intervene as early as possible to reduce the harm to the individual, family and society.
Assuntos
Sintomas Comportamentais/sangue , Proteínas de Ligação a DNA/sangue , Comportamento Problema , Proteínas Proto-Oncogênicas c-akt/sangue , Fatores de Transcrição/sangue , Proteína X Associada a bcl-2/sangue , Adulto , Biomarcadores/sangue , Correlação de Dados , Ajustamento Emocional/fisiologia , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND This study investigated the expression of the BCL2 and BAX mRNA, inflammatory cytokines, interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-alpha (TNF-alpha), and cardiac function in patients with chronic heart failure (CHF). The New York Heart Association (NYHA) Functional Classification and measurement of the left ventricular ejection fraction (LVEF) evaluated cardiac function. MATERIAL AND METHODS Patients with CHF (n=60) due to coronary heart disease, hypertensive heart disease, and cardiomyopathy, and healthy controls (n=30) were studied. Enzyme-linked immunosorbent assay (ELISA) measured serum levels of IL-1ß, IL-6, and TNF-alpha. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) detected mRNA expression of BCL2 and BAX in peripheral blood mononuclear cells (PBMCs). Color Doppler ultrasound measured the LVEF, and the NYHA classification of CHF was used. RESULTS In patients with CHF, levels of IL-1ß, IL-6 and TNF-alpha, and mRNA expression of BAX were significantly increased compared with the control group (p<0.01); BCL2 mRNA level was significantly lower (p<0.01). There were no significant differences in the expression levels of inflammatory cytokines, or BCL2 or BAX mRNA in patients with CHF due to coronary heart disease, hypertensive heart disease, or cardiomyopathy. Expression levels of IL-1ß, IL-6, TNF-alpha, and BAX mRNA were significantly associated with the degree of CHF. Cardiac function was negatively correlated with LVEF (p<0.05). Expression levels of BCL2 mRNA level were negatively correlated with cardiac function (p<0.05), and positively correlated with LVEF (p<0.05). CONCLUSIONS Levels of IL-1ß, IL-6, TNF-alpha, and BAX mRNA were negatively correlated with cardiac function, and BCL2 mRNA expression was positively associated with CHF.
Assuntos
Citocinas/sangue , Regulação da Expressão Gênica , Insuficiência Cardíaca/sangue , Insuficiência Cardíaca/genética , Mediadores da Inflamação/sangue , Leucócitos Mononucleares/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteína X Associada a bcl-2/genética , Idoso , Cardiomiopatias/sangue , Cardiomiopatias/genética , Cardiomiopatias/fisiopatologia , Doença Crônica , Doença das Coronárias/sangue , Doença das Coronárias/genética , Doença das Coronárias/fisiopatologia , Feminino , Insuficiência Cardíaca/fisiopatologia , Testes de Função Cardíaca , Humanos , Hipertensão/sangue , Hipertensão/genética , Hipertensão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-bcl-2/sangue , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Volume Sistólico , Proteína X Associada a bcl-2/sangueRESUMO
BACKGROUND Cardiac remote ischemic conditioning (RIC) is a noninvasive cardioprotective method in ischemia-reperfusion injury and acute myocardial infarction (AMI). The aims of this study were to investigate the effects of RIC in a rat model of AMI. MATERIAL AND METHODS Adult male Sprague-Dawley rats included the AMI group that underwent ligation of the left anterior descending (LAD) coronary artery (n=24), the RIC group that consisted the AMI rat model treated with RIC once daily in the left hind limb until days 1, 7 and 14 (n=24), and the sham group (n=24). Myocardial infarct size was measured by routine histology with triphenyltetrazolium chloride (TTC) and Masson's trichrome histochemical staining for myocardial necrosis and fibrosis, respectively. Serum levels of Bcl-2, Bax, caspase-3, and inducible nitric oxide synthase (iNOS) were measured by enzyme-linked immunosorbent assay (ELISA). The apoptosis index was detected using the TUNEL assay. Spectrophotometry of the myocardium was used to identify mitochondrial complexes and myocardial ATP. RESULTS The RIC group showed improved cardiac hemodynamics, reduced the size of the myocardial infarction, upregulated expression of Bcl-2, and down-regulation of the levels of Bax, caspase-3, and iNOS, and reduced cardiac myocyte apoptosis and inhibited the opening of the mitochondrial permeability transition pore (MPTP). CONCLUSIONS In a rat model of AMI, RIC improved the hemodynamic index, reduce the levels of apoptosis and myocardial injury, and improved mitochondrial function.
Assuntos
Precondicionamento Isquêmico/métodos , Infarto do Miocárdio/metabolismo , Traumatismo por Reperfusão/prevenção & controle , Animais , Apoptose , Cardiotônicos , Caspase 3/análise , Caspase 3/sangue , Modelos Animais de Doenças , Traumatismos Cardíacos/prevenção & controle , Hemodinâmica , Masculino , Mitocôndrias/metabolismo , Infarto do Miocárdio/fisiopatologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Miocárdio/metabolismo , Miocárdio/patologia , Proteínas Proto-Oncogênicas c-bcl-2/análise , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/terapia , Proteína X Associada a bcl-2/análise , Proteína X Associada a bcl-2/sangueRESUMO
The potential benefits of omega-3 fatty acids and branched-chain amino acids (BCAAs) supplements on exercise-induced apoptosis are not clear. In a crossover randomized study, 11 men (age = 62.8 ± 2.2 years) performed an acute bout of resistance exercise and underwent 1-week supplementation with either 20 g of BCAA or 2,700 mg of omega-3/day. Subjects performed the same exercise after supplementation protocols. Following a 3-week washout period, subjects switched groups. Circulating levels of soluble Fas ligand (sFasL), cytochrome c, Bax, Bcl-2, and nuclear factor-kappa B were measured before and immediately after exercise sessions. sFasL, cytochrome c, and Bax increased after exercise. Simple main effect of time on sFasl was significant in control trial but not in omega-3 and BCAA trials. There were no differences in nuclear factor-kappa B and Bcl-2 between control and supplement trials. This study showed that adding omega-3 fatty acids or BCAA to the dietary regime of old men could partially attenuate resistance exercise-induced apoptosis.
Assuntos
Aminoácidos de Cadeia Ramificada/administração & dosagem , Apoptose , Suplementos Nutricionais , Ácidos Graxos Ômega-3/administração & dosagem , Treinamento Resistido , Biomarcadores/sangue , Estudos Cross-Over , Citocromos c/sangue , Proteína Ligante Fas/sangue , Humanos , Masculino , Pessoa de Meia-Idade , NF-kappa B/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteína X Associada a bcl-2/sangueRESUMO
PURPOSE: To investigate the apoptotic mechanisms in rabbits with blast-induced acute lung injury (ALI). METHODS: A total of 40 rabbits were randomly divided into a blank control group (A, n=10) and an experimental group (EXP, n=30). Explosion-induced chest-ALI models were prepared and sampled at different time points (4, 12, and 24h after modeling, T1-T3) to test the lung dry weight/wet weight ratio (W/D) and arterial oxygen pressure (PaO2), apoptosis of lung tissue by the TUNEL assay, and Caspase-3, Bax, and Bcl-2 levels by immunohistochemical analysis. Furthermore, lung tissue was sampled to observe pathological morphology by microscopy. RESULTS: Under a light microscope, Group EXP exhibited obvious edema in the pulmonary interstitial substance and alveoli, a large number of red blood cells, inflammatory cells, and serous exudation in the alveolar cavity, as well as thickening of the pulmonary interstitial fluid. Compared to Group A, the W/D ratio was significantly increased in Group EXP (P<0.01), while PaO2 was significantly reduced (P<0.01). The apoptosis index was significantly increased (P<0.01), and caspase-3 and Bax/Bcl-2 levels were increased (P<0.01). CONCLUSION: Apoptosis plays an important role in the occurrence and development of acute lung injury in rabbits by participating in lung injury and promoting the progression of ALI.
Assuntos
Lesão Pulmonar Aguda/fisiopatologia , Apoptose/fisiologia , Traumatismos por Explosões/fisiopatologia , Lesão Pulmonar Aguda/sangue , Lesão Pulmonar Aguda/patologia , Animais , Traumatismos por Explosões/sangue , Traumatismos por Explosões/patologia , Caspase 3/sangue , Modelos Animais de Doenças , Feminino , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Alvéolos Pulmonares/patologia , Coelhos , Distribuição Aleatória , Proteína X Associada a bcl-2/sangueRESUMO
Abstract Purpose: To investigate the apoptotic mechanisms in rabbits with blast-induced acute lung injury (ALI). Methods: A total of 40 rabbits were randomly divided into a blank control group (A, n=10) and an experimental group (EXP, n=30). Explosion-induced chest-ALI models were prepared and sampled at different time points (4, 12, and 24h after modeling, T1-T3) to test the lung dry weight/wet weight ratio (W/D) and arterial oxygen pressure (PaO2), apoptosis of lung tissue by the TUNEL assay, and Caspase-3, Bax, and Bcl-2 levels by immunohistochemical analysis. Furthermore, lung tissue was sampled to observe pathological morphology by microscopy. Results: Under a light microscope, Group EXP exhibited obvious edema in the pulmonary interstitial substance and alveoli, a large number of red blood cells, inflammatory cells, and serous exudation in the alveolar cavity, as well as thickening of the pulmonary interstitial fluid. Compared to Group A, the W/D ratio was significantly increased in Group EXP (P<0.01), while PaO2 was significantly reduced (P<0.01). The apoptosis index was significantly increased (P<0.01), and caspase-3 and Bax/Bcl-2 levels were increased (P<0.01). Conclusion: Apoptosis plays an important role in the occurrence and development of acute lung injury in rabbits by participating in lung injury and promoting the progression of ALI.
Assuntos
Animais , Masculino , Feminino , Coelhos , Traumatismos por Explosões/fisiopatologia , Apoptose/fisiologia , Lesão Pulmonar Aguda/fisiopatologia , Alvéolos Pulmonares/patologia , Traumatismos por Explosões/patologia , Traumatismos por Explosões/sangue , Distribuição Aleatória , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Modelos Animais de Doenças , Proteína X Associada a bcl-2/sangue , Caspase 3/sangue , Lesão Pulmonar Aguda/patologia , Lesão Pulmonar Aguda/sangueRESUMO
OBJECTIVE: Strenuous exercise can induce apoptosis in a variety of tissues. We investigated the effects of creatine loading on apoptosis markers after downhill running. DESIGN: Twenty-two middle-aged men were randomly assigned to either a creatine or a placebo group. Crossover design, double-blind controlled supplementation was performed using 20 g/d(-1) of creatine or maltodextrin for 7 days. Downhill running (12% incline) at 70% of heart rate maximum for 40 mins was performed on the eighth day. Blood samples were taken on the day before supplementation, after supplementation and after running. RESULTS: There were no significant changes in the caspase-3, caspase-9, p53, Bax, and IGF-1 concentrations from presupplementation to postsupplementation in both groups of creatine and placebo (P > 0.05). There were significant increases (P < 0.05) in serum caspase-3, caspase-9, p53, and Bax after running in the placebo group. These markers were not noticeably changed in the creatine group (P > 0.05). Bcl-2 was unchanged in the placebo group but substantially increased (P < 0.05) in the creatine group. No significant changes were observed in IGF-1 concentration after running comparing to prerunning in both groups (P > 0.05). Lactate levels increased similarly in both groups (P < 0.05). CONCLUSIONS: The findings indicate that creatine supplementation could prevent exercise-induced apoptotic markers.
Assuntos
Apoptose/fisiologia , Creatina/farmacologia , Suplementos Nutricionais , Corrida/fisiologia , Biomarcadores/sangue , Caspase 3/sangue , Caspase 9/sangue , Estudos Cross-Over , Método Duplo-Cego , Genes bcl-2 , Genes p53 , Humanos , Fator de Crescimento Insulin-Like I/metabolismo , Ácido Láctico/sangue , Masculino , Pessoa de Meia-Idade , Proteína X Associada a bcl-2/sangueRESUMO
PURPOSE: The study was processed to investigate the effect of astaxanthin (AST; 3,3-dihydroxybeta, beta-carotene-4,4-dione) on the acute kidney injury induced by iohexol and the relationship with SIRT1/FOXO3a signal pathway. METHODS: Thirty male Sprague Dawley rats were randomly divided into five groups as follows: control group (CON; olive oil only), contrast media group, astaxanthin control group (100 mg/kg), low astaxanthin dose group (LAG, 50 mg/kg) and high astaxanthin dose group (HAG, 100 mg/kg). As followed, serum creatinine (SCr), blood urea nitrogen (BUN), the oxidative stress markers and apoptosis-related proteins were detected. Human proximal tubular epithelial cells (HK-2) were cultured in DMEM/F12 medium in vitro and then randomly divided into appropriate experimental groups: normal group (N), dimethyl sulfoxide (DMSO), iohexol group (I), iohexol + (1.0, 10.0 µmol/L) astaxanthin group (I + LAST; I + HAST), iohexol + SIRT1 inhibitors (nicotinamide) group (NA) and iohexol + si-RNA FOXO3a group (si-RNA FOXO3a); when cultured for 24 h, cell proliferation ability was tested by cell counting kit (CCK-8), reactive oxygen species (ROS) were detected by flow cytometry and the expression of SIRT1 and FOXO3a were observed using western blot. RESULTS: At the end of the experiment, the levels of SCr, BUN and malondialdehyde (MDA) were all increased in the CM group. The LAG and HAG reduce superoxide anion (SOD) activity, catalase (CAT) activity, glutathione peroxidase (GPx) activity and glutathione (GSH) content, as well as SCr and BUN level. Moreover, apoptosis-associated proteins, caspase 3 p17, bax and bcl-2 were upregulated. In HK-2 cells, after adding iohexol, proliferation and intracellular ROS levels were significantly increased. Using astaxanthin in advance after the intervention, the result is opposite. SIRTl inhibitors NA can reduce the expression of SIRTl and decrease the expression of FOXO3a protein. Si-RNA FOXO3a reduces the expression of FOXO3a but had no significant effect on the expression of SIRT1. CONCLUSIONS: Our study demonstrates that the intervention of astaxanthin could attenuate the oxidative stress and apoptosis in contrast-induced acute kidney injury (CI-AKI), and the SIRT1/FOXO3a pathway participates in the contrast-induced apoptosis of HK-2 cells. Finally, astaxanthin reduces CI-AKI by suppression of apoptosis, which may be through inhibition of SIRT1/FOXO3a pathways.
Assuntos
Injúria Renal Aguda/tratamento farmacológico , Proteína Forkhead Box O3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/metabolismo , Injúria Renal Aguda/sangue , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/patologia , Animais , Apoptose/efeitos dos fármacos , Nitrogênio da Ureia Sanguínea , Caspase 3/sangue , Catalase/sangue , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Meios de Contraste/efeitos adversos , Creatinina/sangue , Células Epiteliais , Proteína Forkhead Box O3/genética , Glutationa/sangue , Glutationa Peroxidase/sangue , Humanos , Iohexol/efeitos adversos , Túbulos Renais Proximais/citologia , Masculino , Malondialdeído/sangue , Niacinamida/farmacologia , Peptídeos/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , RNA Interferente Pequeno/farmacologia , Ratos , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/sangue , Complexo Vitamínico B/farmacologia , Xantofilas/farmacologia , Xantofilas/uso terapêutico , Proteína X Associada a bcl-2/sangueRESUMO
To investigate the role of S100B, oxidative stress and the apoptosis signaling pathways in the sevoflurane induced neuroprotective effect on stroke. The brain injury, molecular and cellular damage, and functional recovery were investigated upon ischemic brain injury followed by sevoflurane treatment. Longa rodent stroke scales was used to quantify neurological deficits. TTC staining was used to measure infarct volume of brain tissue. Absolute brain water content was measured by wet/dry weight method. The neuronal morphological change was assessed by H and E staining. The spatial learning and memory ability were measured by water maze test. Serum proteins including S100B, GSH-PX, SOD, Bcl-2, Bax, Caspase-3 were measured by ELISA. The level of NOS and NO in serum was determined by colorimetric method. Compared with control, the serum proteins including S100B, Bax, NO, Caspase-3, and NOS activity in cerebral infarction rats increased significantly while SOD, GSH-PX, and Bcl-2 decreased significantly. Diabetic mellitus complicated with cerebral infarction rats showed more dramatic increase for S100B, Bax, NO, Caspase-3, and NOS activity and dramatic decrease for SOD, GSH-PX, and Bcl-2. Interestingly, sevoflurane reduced the changes significantly. The S100B level positively correlated with brain damage, NO, Bax, caspase-3, and NOS activity but negatively correlated with SOD, Bax, and GSH-PX. Brain damage in sevoflurane groups decreased while behavior outcomes including Longa neurologic score, learning, and memory increased significantly. The neuroprotective effect of sevoflurane is associated with defense mechanisms against free radical-induced oxidative stress and inhibition of apoptosis. S100B protein correlated with oxidative stress and the apoptosis signaling pathways.
Assuntos
Lesões Encefálicas/tratamento farmacológico , Isquemia Encefálica/tratamento farmacológico , Diabetes Mellitus Experimental/tratamento farmacológico , Subunidade beta da Proteína Ligante de Cálcio S100/sangue , Sevoflurano/administração & dosagem , Animais , Apoptose/efeitos dos fármacos , Lesões Encefálicas/sangue , Lesões Encefálicas/patologia , Isquemia Encefálica/sangue , Isquemia Encefálica/patologia , Caspase 3/sangue , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/patologia , Dieta , Modelos Animais de Doenças , Humanos , Neurônios/efeitos dos fármacos , Neurônios/patologia , Fármacos Neuroprotetores/administração & dosagem , Estresse Oxidativo/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Ratos , Transdução de Sinais/efeitos dos fármacos , Acidente Vascular Cerebral/patologia , Acidente Vascular Cerebral/prevenção & controle , Superóxido Dismutase-1/sangue , Proteína X Associada a bcl-2/sangueRESUMO
Platelets are anucleate cells, known for their pivotal roles in hemostasis, inflammation, immunity, and disease progression. Being anuclear, platelets are known to express several transcriptional factors which exert nongenomic functions, including the positive and negative regulation of platelet activation. NF-κB is one such transcriptional factor involved in the regulation of genes for survival, proliferation, inflammation and immunity. Although, the role NF-κB in platelet activation and aggregation is partially known, its function in management of platelet survival and apoptosis remain unexplored. Therefore, two unrelated inhibitors of NF-κB activation, BAY 11-7082 and MLN4924 were used to determine the role of NF-κB in platelets. Inhibition of NF-κB caused decreased SERCA activity and increased cytosolic Ca2+ level causing ER stress which was determined by the phosphorylation of eIF2-α. Further, there was increased BAX and decreased BCl-2 levels, incidence of mitochondrial membrane potential depolarization, release of cytochrome c into cytosol, caspase activation, PS externalization and cell death in BAY 11-7082 and MLN4924 treated platelets. The obtained results demonstrate the critical role played by NF-κB in Ca2+ homeostasis and survival of platelets. In addition, the study demonstrates the potential side effects associated with NF-κB inhibitors employed during inflammation and cancer therapy.
Assuntos
Apoptose/fisiologia , Plaquetas/citologia , Plaquetas/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , NF-kappa B/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Plaquetas/efeitos dos fármacos , Cálcio/sangue , Ciclopentanos/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , NF-kappa B/sangue , Nitrilas/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Pirimidinas/farmacologia , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/sangue , Sulfonas/farmacologia , Proteína X Associada a bcl-2/sangueRESUMO
Background: Anucleate platelets can undergo apoptosis in response to various stimuli, as do nucleated cells. Cardiopulmonary bypass (CPB) causes platelet dysfunction and can also activate platelet apoptotic pathways. We therefore evaluated time-dependent changes in blood platelet Bax (a pro-apoptotic molecule) levels and platelet dysfunction after cardiac surgery. Methods: We assessed blood samples obtained from subjects having on-pump or off-pump coronary artery bypass graft surgery ( n =20 each). We also evaluated the in vitro effects of platelet Bax increase in eight healthy volunteers. Results: Thrombin-induced platelet calcium mobilisation and platelet-surface glycoprotein Ib (GPIb) expression were lowest at weaning from CPB and did not recover on postoperative day one. On-pump surgery increased platelet expression of Bax, especially the oligomerised form, along with translocation of Bax from the cytosol to mitochondria and platelet-surface tumour necrosis factor-alpha (TNF-α)-converting enzyme (TACE) expression. In contrast, mitochondrial cytochrome c expression was reduced. While similar in direction, the magnitude of the observed changes was smaller in patients having off-pump surgery. In vitro , a cell-permeable Bax peptide increased platelet Bax expression to the same extent seen during bypass and produced similar platelet changes. These apoptotic-like changes were largely reversed by Bcl-xL pre-administration, and were completely reversed by combined application of inhibitors that stabilise outer mitochondrial membrane permeability and TACE. Conclusions: CPB increases platelet Bax expression, which contributes to reduced platelet-surface GPIb expression and thrombin-induced platelet calcium changes. These changes in platelet apoptotic signalling might contribute to platelet dysfunction after CPB. Clinical trial registration: UMIN Clinical Trials Registry (number UMIN000006033).
Assuntos
Plaquetas/patologia , Ponte Cardiopulmonar , Complicações Pós-Operatórias/sangue , Trombina/farmacologia , Proteína X Associada a bcl-2/sangue , Idoso , Idoso de 80 Anos ou mais , Apoptose , Western Blotting , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Ativação Plaquetária , Agregação Plaquetária , Complicações Pós-Operatórias/patologia , Estudos Prospectivos , Proteína X Associada a bcl-2/genéticaRESUMO
BACKGROUND: Stress is a significant factor in the etiology of depression. Stellate ganglion block (SGB) has been shown to maintain the stability of the autonomic system and to affect the neuroendocrine system, including the hypothalamic-pituitary-adrenal (HPA) axis. The objective of this study was to determine the antidepressant-like effects of SGB on the autonomic system and the HPA axis, apoptosis-related proteins, related spatial learning and memory impairment, and sensorimotor dysfunction. METHODS: Forty-eight Sprague Dawley rats were assigned to four experimental groups: control + saline (sham group), control + SGB (SGB group), unpredictable chronic mild stress (UCMS) + saline (UCMS group), and UCMS + SGB (UCSG group). Stress-induced effects and the function of SGB were assessed using measures of body weight, coat state, sucrose consumption, and behavior in open-field and Y-maze tests. Neuronal damage was assessed histologically using the hematoxylin-eosin (HE) staining method, while western blotting was used to investigate changes in the expression of apoptosis-related proteins. Plasma corticotropin-releasing factor (CRF), adrenocorticotropic hormone (ACTH), corticosterone (CORT), noradrenaline and adrenaline were measured to evaluate changes in the autonomic system and HPA axis. RESULTS: SGB treatment significantly improved sensorimotor dysfunction and spatial learning and memory impairment following UCMS. Moreover, UCMS significantly decreased body weight, sucrose preference and anti-apoptotic protein Bcl-2, and increased scores on measures of coat state, adrenal gland weight, levels of CORT, CRF, ACTH, noradrenaline and adrenaline, as well as increased neuronal loss, cell shrinkage, nuclear condensation, and the pro-apoptotic protein Bax. These symptoms were attenuated by treatment with SGB. CONCLUSIONS: These findings suggest that SGB can attenuate depression-like behaviors induced by chronic stress. These protective effects appear to be due to an anti-apoptotic mechanism of two stress pathways-the autonomic system and the HPA axis.