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1.
Neurosci Lett ; 744: 135615, 2021 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-33421493

RESUMO

BACKGROUND: Spinal cord ischemia/reperfusion injury is a common clinical, pathophysiological phenomenon with complex molecular mechanisms. Currently, there are no therapeutics available to alleviate the same. This study investigates the protective effects of sulfiredoxin-1 (Srxn 1) on spinal cord neurons following exposure to oxygen-glucose deprivation/reoxygenation (OGD/R) treatment. MATERIALS AND METHODS: Primary spinal cord neurons were cultured, detected by anti-tubulin ßⅢ, and transfected with adeno-associated virus (AAV)-Srxn 1 to overexpress Srxn 1. They were identified by their morphology and CCK-8 assay. The superoxide dismutase level was measured by superoxide dismutase assay. Malondialdehyde level was measured by malondialdehyde assay. The apoptosis ratio was calculated by Hoechst 33342 and Annexin V-PE/7-AAD staining. Mitochondrial transmembrane potential (Δψm) was detected by tetramethylrhodamine-methyl ester-perchlorate (TMRM) staining. The mRNA expression levels of Srxn 1 and caspase 3 were detected by quantitative reverse transcription-polymerase chain reaction, and the protein expression levels of Srxn 1, bax, bcl-2, cytosolic cytochrome c, and caspase 3 were detected by western blotting. RESULTS: AAV-Srxn 1 up-regulated mRNA and protein levels of Srxn 1 in spinal cord neurons. Following exposure to OGD/R, overexpression of Srxn 1 improved the neuronal viability, alleviated the neuron apoptosis, enhanced the mitochondrial transmembrane potential, increased the SOD level, decreased the MDA level, inhibited the expression of cytosolic cytochrome c, bax, and caspase 3, and promoted the expression of bcl-2. CONCLUSION: Srxn 1 plays a significant role in anti-apoptosis of spinal cord neurons, and Srxn 1 may be a potential therapeutic target for spinal cord I/R injury.


Assuntos
Caspase 3/biossíntese , Citocromos c/sangue , Neurônios/metabolismo , Estresse Oxidativo/fisiologia , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/biossíntese , Proteína X Associada a bcl-2/biossíntese , Animais , Apoptose/fisiologia , Hipóxia Celular/fisiologia , Citocromos c/antagonistas & inibidores , Glucose/deficiência , Oxigênio/metabolismo , Ratos , Transdução de Sinais/fisiologia , Medula Espinal/metabolismo , Proteína X Associada a bcl-2/antagonistas & inibidores
2.
Sci Rep ; 10(1): 5109, 2020 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-32198370

RESUMO

Splenic immune function was enhanced in diet-induced-obese (DIO) mice caused by Escherichia coli. The changes in spleen function on apoptosis were still unknown. Two hundred mice in groups Lean-E. coli and DIO-E. coli were intranasal instillation of E. coli. And another two hundred mice in groups Lean-PBS and DIO-PBS were given phosphate-buffered saline (PBS). Subsequently, spleen histology was analyzed. Then the rates of spleen cell (SC) apoptosis, and expression of the genes and proteins of Bcl-2, Bax, caspase-3 and caspase-9 were quantified in each group at 0 h (uninfected), 12 h, 24 h, and 72 h postinfection. The SC apoptosis rates of the DIO-E. coli groups were lower than those of the DIO-PBS groups at 12, 24 and 72 h (p < 0.05). Anti-apoptotic Bcl-2 expression gene and protein of the DIO-E. coli groups were higher than those of the DIO-PBS groups (p < 0.05). Gene expressions of pro-apoptotic Bax, caspase-3 and caspase-9 of the DIO-E. coli groups were lower than those of DIO-PBS groups at 12, 24 and 72 h (p < 0.05). The SC apoptosis rates of the Lean-E. coli groups were higher than those of the Lean- PBS groups at 12 h and 24 h (p < 0.05). Interestingly, the SC apoptosis rates in the DIO-E. coli groups were lower than those of the Lean-E. coli groups at 12 h (p < 0.05). In conclusion, our results suggested that the DIO mice presented stronger anti-apoptotic abilities than Lean mice in non-fatal acute pneumonia induced by E. coli infection, which is more conducive to protecting the spleen and improving the immune defense ability of the body.


Assuntos
Apoptose/imunologia , Infecções por Escherichia coli/imunologia , Escherichia coli/imunologia , Obesidade/imunologia , Pneumonia/imunologia , Baço/imunologia , Administração Intranasal , Animais , Caspase 3/biossíntese , Caspase 9/biossíntese , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/mortalidade , Macrófagos/imunologia , Masculino , Megacariócitos/imunologia , Camundongos , Camundongos Obesos , Neutrófilos/imunologia , Pneumonia/microbiologia , Pneumonia/mortalidade , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Baço/citologia , Proteína X Associada a bcl-2/biossíntese
3.
Parasitol Res ; 119(1): 55-62, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31786697

RESUMO

In developing countries, low-cost control and treatment programs that offer combined approaches against diseases and their vectors are certainly needed. Ivermectin (IVM) has been well known for its role in the treatment of parasitic diseases, due to its effect on glutamate-gated chloride channels. These same channels are also present in the mosquito vector, and thus, research has focused on the insecticidal effects of this drug. Possible alternative mechanisms of IVM on the physiology of mosquitoes, however, have not been sufficiently elaborated. We assessed the protease activity, lipid peroxidation, and local expression of STAT, p53, caspase-3, and Bax markers to study the effect of this antibiotic on digestion and immunity in Culex pipiens. Sugar- and blood-feeding assays were employed to investigate the potential influence of blood feeding on the dynamics of these parameters. IVM was found to have an effect on protease activity, lipid peroxidation as well as the expression of different markers investigated in this work. The focus on the detailed effect of this drug certainly opens the gate to broadening the spectrum of IVM and expanding its health and economic benefit, especially that it is relatively more affordable than other antibiotics on the market.


Assuntos
Canais de Cloreto/efeitos dos fármacos , Culex/efeitos dos fármacos , Inseticidas/farmacologia , Ivermectina/farmacologia , Animais , Caspase 3/biossíntese , Culex/imunologia , Culex/fisiologia , Digestão/efeitos dos fármacos , Imunidade/efeitos dos fármacos , Proteínas de Insetos/biossíntese , Mosquitos Vetores/efeitos dos fármacos , Mosquitos Vetores/fisiologia , Fatores de Transcrição STAT/biossíntese , Proteína Supressora de Tumor p53/biossíntese , Proteína X Associada a bcl-2/biossíntese
4.
Neurotox Res ; 37(2): 356-365, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31493121

RESUMO

Paraoxon is the bioactive metabolite of organophosphate (OP) pesticide, parathion. This study aimed to evaluate the expression of apoptosis-related genes and histopathological changes in rat prefrontal cortex following exposure to three different doses of paraoxon. Paraoxon (0.3, 0.7, or 1 mg/kg) or corn oil (vehicle) were intraperitoneally injected to adult male Wistar rats. After 14 or 28 days, mRNA and protein levels of Bax, Bcl-2, and caspase-3 were measured in prefrontal cortex using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and western blotting, respectively. In addition, neuronal injury and astrocyte activation were assessed using cresyl violet staining and glial fibrillary acidic protein (GFAP) immune-positive cells, respectively. Treatment with 0.7 and 1 mg/kg of paraoxon increased mRNA and protein levels of Bax and caspase-3 at 14 and 28 days post-exposure, while mRNA and protein levels of Bcl-2 decreased only in 1 mg/kg group after 14 days. Furthermore, a significant decrease in the number of neurons and a significant increase in the number of GFAP-positive cells were observed in rats receiving 0.7 and 1 mg/kg of paraoxon at both time points. Collectively, our results showed that apoptosis is a major mechanism for neuronal damage after exposure to paraoxon. Also, paraoxon-induced neuronal loss was correlated with activation of astrocytes. Since paraoxon-induced neuronal damage is closely related to convulsion, clinical management of convulsion could protect neuronal brain damage.


Assuntos
Apoptose/efeitos dos fármacos , Astrócitos/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Paraoxon/toxicidade , Córtex Pré-Frontal/efeitos dos fármacos , Animais , Apoptose/fisiologia , Astrócitos/metabolismo , Caspase 3/biossíntese , Caspase 3/genética , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Inibidores da Colinesterase/toxicidade , Expressão Gênica , Masculino , Neurônios/metabolismo , Córtex Pré-Frontal/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/genética , Ratos , Ratos Wistar , Proteína X Associada a bcl-2/biossíntese , Proteína X Associada a bcl-2/genética
5.
Acta Neurobiol Exp (Wars) ; 79(4): 352-366, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31885392

RESUMO

Methylphenidate (MPH) abuse causes adverse neurobehavioral and neurochemical effects. Some herbal components such as crocin have shown neuroprotective properties. The current study evaluates the potential role of the cyclic AMP response element binding protein (CREB)­brain­derived neurotrophic factor (BDNF) signaling pathway in mediating the neuroprotective effects of crocin against MPH­induced neurotoxicity in rats. Seventy adult male rats were randomly divided into seven groups. Group 1 and 2 received 0.7 ml/rat of normal saline and 10 mg/kg of MPH, respectively. Groups 3, 4, 5, and 6 were treated simultaneously with MPH (10 mg/kg) and crocin (10, 20, 40, and 80 mg/kg, respectively) for 21 days. Group 7 was treated with crocin (80 mg/kg) alone for 21 days. The Morris water maze (MWM) and open field test were used to assess cognitive and locomotor activities. Hippocampal neurotoxicity parameters and levels of BDNF and CREB were evaluated. Simultaneous treatment with various doses of crocin reduced the MPH­induced cognition disturbances and hyperlocomotion. In addition, lipid peroxidation increased with MPH treatment and levels of the oxidized forms of glutathione (GSSG), interleukin 1 beta (IL­1ß), tumor necrosis factor alpha (TNF­α), and Bax increased. MPH treatment decreased levels of the reduced form of glutathione (GSH), P­CREB, Bcl­2, and BDNF in the hippocampus. MPH also reduced activity of superoxide dismutase, glutathione peroxidase, and glutathione reductase in the hippocampus. In contrast, crocin attenuated MPH­induced oxidative stress, inflammation, and apoptosis, and increased levels of P­CREB and BDNF. Thus, crocin - likely via stimulation of the P­CREB/BDNF signaling pathway - displayed neuroprotection against MPH­induced neurotoxicity.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/fisiologia , Carotenoides/uso terapêutico , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/fisiologia , Hipocampo/efeitos dos fármacos , Intoxicação por MPTP/tratamento farmacológico , Proteínas do Tecido Nervoso/fisiologia , Fármacos Neuroprotetores/uso terapêutico , Transdução de Sinais/efeitos dos fármacos , Animais , Fator Neurotrófico Derivado do Encéfalo/biossíntese , Fator Neurotrófico Derivado do Encéfalo/genética , Carotenoides/farmacologia , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/biossíntese , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Comportamento Exploratório , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Hipocampo/metabolismo , Peroxidação de Lipídeos , Masculino , Aprendizagem em Labirinto , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/genética , Fármacos Neuroprotetores/farmacologia , Oxirredução , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/genética , Distribuição Aleatória , Ratos , Ratos Wistar , Transdução de Sinais/fisiologia , Superóxido Dismutase/metabolismo , Natação , Proteína X Associada a bcl-2/biossíntese , Proteína X Associada a bcl-2/genética
6.
CNS Neurol Disord Drug Targets ; 18(10): 791-797, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31686636

RESUMO

BACKGROUND: Our previous study has indicated that somatostatin potently inhibits neuropathic pain through the activation of its type 2 receptor (SSTR2) in mouse dorsal root ganglion and spinal cord. However, the underlying mechanism of this activation has not been elucidated clearly. OBJECTIVE: The aim of this study is to perform the pharmacological studies on the basis of sciatic nerve-pinch mice model and explore the underlying mechanism involving SSTR2. METHODS: On the basis of a sciatic nerve-pinch injury model, we aimed at comparing the painful behavior and dorsal root ganglion neurons neurochemical changes after the SSTR2 antibody (anti- SSTR2;5µl,1µg/ml) administration in the mouse. RESULTS: After pinch nerve injury, we found that the mechanical hyperalgesia and severely painful behavior (autotomy) were detected after the application of SSTR2 antibody (anti-SSTR2; 5µl, 1µg/ml) on the pinch-injured nerve. The up-regulated phosphorylated ERK (p-ERK) expression and the apoptotic marker (i.e., Bax) were significantly decreased in DRGs after anti-SSTR2 treatment. CONCLUSION: The current data suggested that inhibitory changes in proteins from the apoptotic pathway in anti-SSTR2-treated groups might be taking place to overcome the protein deficits caused by SSTR2 antibody and supported the new therapeutic intervention with SSTR2 antagonist for neuronal degeneration following nerve injury.


Assuntos
Anticorpos/imunologia , Gânglios Espinais/fisiopatologia , Hiperalgesia/prevenção & controle , Hiperalgesia/fisiopatologia , Receptores de Somatostatina/imunologia , Receptores de Somatostatina/fisiologia , Animais , MAP Quinases Reguladas por Sinal Extracelular/biossíntese , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/metabolismo , Hiperalgesia/metabolismo , Hiperalgesia/psicologia , Masculino , Camundongos , Nervo Isquiático/lesões , Proteína X Associada a bcl-2/biossíntese
7.
C R Biol ; 342(5-6): 186-191, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31495738

RESUMO

The aim of our study was to understand the role of transcription factor p53 in the control of healthy human ovarian cell functions. Ovarian granulosa cells were transfected with a cDNA construct encoding p53. The intracellular accumulation of p53, of the apoptosis marker bax, and of the proliferation marker PCNA, as well as the release of progesterone (P4), insulin-like growth factor I (IGF-I), oxytocin (OT), and prostaglandin F (PGF) and E2 (PGE) were evaluated by quantitative immunocytochemistry and RIA/IRMA. Transfection with the p53 cDNA construct resulted in the accumulation of p53 and bax, in a reduced level of released PCNA and PGF, and in an increased PGE output. No changes in P4, IGF-I, and OT secretion were found. These observations are the first demonstration of the involvement of p53 in the control of healthy human ovarian cell functions, namely, in the downregulation of proliferation, in the upregulation of apoptosis, and in the alteration of PGF and PGE release, but not of P4, IGF-I, or OT.


Assuntos
Ovário/fisiologia , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/fisiologia , Eletroforese em Gel de Poliacrilamida , Feminino , Proteínas de Fluorescência Verde/genética , Humanos , Fator de Crescimento Insulin-Like I/biossíntese , Ovário/metabolismo , Ocitocina/biossíntese , Antígeno Nuclear de Célula em Proliferação/biossíntese , Prostaglandinas F/biossíntese , Proteína X Associada a bcl-2/biossíntese
8.
BMC Neurosci ; 20(1): 41, 2019 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-31387531

RESUMO

BACKGROUND: Oxidative stress and neuroinflammation are central pathogenic mechanisms common to many neurological diseases. Isoliquiritigenin (ISL) is a flavonoid in licorice with multiple pharmacological properties, including anti-inflammatory activity, and has demonstrated protective efficacy against acute neural injury. However, potential actions against cognitive impairments have not been examined extensively. We established a rat model of cognitive impairment by intracerebroventricular injection of lipopolysaccharide (LPS), and examined the effects of ISL pretreatment on cognitive function, hippocampal injury, and hippocampal expression of various synaptic proteins, antioxidant enzymes, pro-inflammatory cytokines, and signaling factors controlling anti-oxidant and pro-inflammatory responses. RESULTS: Rats receiving LPS alone demonstrated spatial learning deficits in the Morris water maze test as evidenced by longer average escape latency, fewer platform crossings, and shorter average time in the target quadrant than untreated controls. ISL pretreatment reversed these deficits as well as LPS-induced decreases in the hippocampal expression levels of synaptophysin, postsynaptic density-95, brain-derived neurotrophic factor, superoxide dismutase, glutathione peroxidase, and BCL-2. ISL pretreatment also reversed LPS-induced increases in TUNEL-positive (apoptotic) cells, BAX/BCL-2 ratio, and expression levels of tumor necrosis factor-α, interleukin (IL)-1ß, IL-6, and C-C motif chemokine ligand 3. Pretreatment with ISL increased the expression levels of phosphorylated (p)-GSK-3ß, nuclear NRF2, HO-1 mRNA, and NQO1 mRNA, and reversed LPS-induced nuclear translocation of nuclear factor (NF)-κB. CONCLUSIONS: ISL protects against LPS-induced cognitive impairment and neuronal injury by promoting or maintaining antioxidant capacity and suppressing neuroinflammation, likely through phosphorylation-dependent inactivation of GSK-3ß, enhanced expression of NRF2-responsive antioxidant genes, and suppression of NF-κB-responsive pro-inflammatory genes.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Chalconas/farmacologia , Disfunção Cognitiva/prevenção & controle , Animais , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Fator Neurotrófico Derivado do Encéfalo , Disfunção Cognitiva/induzido quimicamente , Citocinas/biossíntese , Proteína 4 Homóloga a Disks-Large/biossíntese , Glutationa Peroxidase/biossíntese , Hipocampo/metabolismo , Lipopolissacarídeos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Ratos , Superóxido Dismutase/biossíntese , Sinaptofisina/biossíntese , Proteína X Associada a bcl-2/biossíntese
9.
Drug Dev Res ; 80(7): 933-947, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31343754

RESUMO

Novel 1,1-disubstituted cyclohexane-1-carboxamides 6a-h, 7a-e, and 8a-b were designed and synthesized as apoptotic inducers. Cytotoxicity test revealed that some compounds have strong to moderate effect, while others displayed weak action against different cancer cell lines including, MCF-7, HepG2, A549, and HTC-116. A549 carcinoma cell line exhibited higher sensitivity toward all synthesized candidates especially compounds 6a and 8a which offered the lowest IC50 values 3.03 and 5.21 µM, respectively, relative to the positive control doxorubicin with IC50 value of 3.01 µM. Compared to doxorubicin treatment, compounds 6a and 8a induced caspases-3, -8, and -9 activities and G2/M growth arrest in A549 carcinoma cell line. The expression levels of p53 (tumor suppressor protein that in humans is encoded by the TP53 gene), Bax (apoptosis regulator protein in humans that is encoded by bax gene), and the Bax/Bcl-2 ratio were all higher than those in doxorubicin-treated cells (Bcl-2, B-cell lymphoma 2, encoded in humans by the Bcl-2 gene). Additionally, compounds 6a and 8a appeared to exhibit higher selectivity against MCF-10 human breast normal cell line. The synthesized congeners could be considered as potent apoptotic inducers interfering with extrinsic and intrinsic apoptotic pathways. Moreover, compound 6a was able to form complex with zinc ions as indicated by UV spectrophotometry which revealed its ability for being caspase activator. Molecular docking studies expected the interactions and binding modes of the synthesized inhibitors in the caspase-3 active site.


Assuntos
Apoptose/efeitos dos fármacos , Caspases/metabolismo , Cicloexanos/síntese química , Cicloexanos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Indução Enzimática/efeitos dos fármacos , Simulação de Acoplamento Molecular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Quelantes/química , Cicloexanos/química , Doxorrubicina/farmacologia , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Relação Estrutura-Atividade , Proteína Supressora de Tumor p53/biossíntese , Zinco/química , Proteína X Associada a bcl-2/biossíntese
10.
Med Sci Monit ; 25: 4583-4589, 2019 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-31257361

RESUMO

BACKGROUND The apoptosis of corneal epithelial cells participates in the pathological processes of dry eye, which is expected to be a treatment target for dry eye. The aim of this study was to investigate the effects of vitamin A (VA) on apoptosis of corneal epithelial cells in a mouse model with dry eye induced by benzalkonium chloride (BAC). MATERIAL AND METHODS We randomly divided 60 male BALB/c mice aged 8-10 weeks into 3 groups: the blank control group, the dry eye + vehicle group, and the dry eye + drug group. On the 7th day after the dry eye model successfully induced, the mouse eyeballs removed, and the mouse corneal tissues were isolated. The expression levels of Bax and Bcl-2 in corneal tissues were detected via reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The apoptotic corneal epithelial cells were quantified using terminal deoxynucleotidyl transferase (TdT) deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) staining technique. RESULTS VA suppressed the upregulation of the Bax gene at the mRNA and protein levels, and upregulated the expression of the Bcl-2 gene (P<0.05). TUNEL results revealed that the number of apoptotic epithelial cells in the dry eye group was 40 times larger as that in the blank control group. After the intervention of VA at an appropriate concentration, the number of apoptotic corneal epithelial cells was remarkably reduced to about 10 times that in the blank control group (P<0.05). CONCLUSIONS VA can inhibit upregulation of the expressions of Bax and Bcl-2 in the epithelial cells of mice with dry eye induced by BAC, so as to suppress the apoptosis of epithelial cells in mice with dry eye.


Assuntos
Proteínas Proto-Oncogênicas c-bcl-2/genética , Vitamina A/farmacologia , Proteína X Associada a bcl-2/genética , Animais , Apoptose/efeitos dos fármacos , Compostos de Benzalcônio/farmacologia , Córnea/efeitos dos fármacos , Córnea/metabolismo , Córnea/patologia , Modelos Animais de Doenças , Síndromes do Olho Seco/tratamento farmacológico , Síndromes do Olho Seco/metabolismo , Células Epiteliais/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteína X Associada a bcl-2/biossíntese
11.
Oxid Med Cell Longev ; 2019: 4578175, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31210840

RESUMO

NADPH oxidase (Nox) is considered a major source of reactive oxygen species (ROS) in the heart in normal and pathological conditions. However, the role of Nox in severe acute pancreatitis- (SAP-) associated cardiac injury remains unclear. Therefore, we aim to investigate the contribution of Nox to SAP-associated cardiac injury and to explore the underlying molecular mechanisms. Apocynin, a Nox inhibitor, was given at 20 mg/kg for 30 min before SAP induction by a retrograde pancreatic duct injection of 5% sodium taurocholate. Histopathological staining, Nox activity and protein expression, oxidative stress markers, apoptosis and associated proteins, cardiac-related enzyme indexes, and cardiac function were assessed in the myocardium in SAP rats. The redox-sensitive MAPK signaling molecules were also examined by western blotting. SAP rats exhibited significant cardiac impairment along with increased Nox activity and protein expression, ROS production, cell apoptosis, and proapoptotic Bax and cleaved caspase-3 protein levels. Notably, Nox inhibition with apocynin prevented SAP-associated cardiac injury evidenced by a decreased histopathologic score, cardiac-related enzymes, and cardiac function through the reduction of ROS production and cell apoptosis. This protective role was further confirmed by a simulation experiment in vitro. Moreover, we found that SAP-induced activation in MAPK signaling molecules in cardiomyocytes was significantly attenuated by Nox inhibition. Our data provide the first evidence that Nox hyperactivation acts as the main source of ROS production in the myocardium, increases oxidative stress, and promotes cell apoptosis via activating the MAPK pathway, which ultimately results in cardiac injury in SAP.


Assuntos
Apoptose , Regulação Enzimológica da Expressão Gênica , Cardiopatias/enzimologia , Miocárdio/enzimologia , NADPH Oxidases/metabolismo , Pancreatite/enzimologia , Espécies Reativas de Oxigênio/metabolismo , Animais , Caspase 3/biossíntese , Modelos Animais de Doenças , Cardiopatias/etiologia , Cardiopatias/patologia , Sistema de Sinalização das MAP Quinases , Masculino , Miocárdio/patologia , NADPH Oxidases/antagonistas & inibidores , Pancreatite/induzido quimicamente , Pancreatite/complicações , Pancreatite/patologia , Ratos , Índice de Gravidade de Doença , Proteína X Associada a bcl-2/biossíntese
12.
Acta Haematol ; 142(4): 208-216, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31163428

RESUMO

BACKGROUND: miR-886-5p plays an important role in many tumors, but it has been rarely investigated in multiple myeloma (MM). We studied the expression of miR-886-5p in the plasma of MM patients and in MM cell lines, and evaluated its biological function to identify its potential involvement in MM. METHODS: We recruited 16 subjects including 10 newly diagnosed MM patients who had not received treatment and 6 healthy individuals. The expression of miR-886-5p in plasma and MM cell lines was examined by quantitative reverse transcription polymerase chain reaction. Cell Counting Kit-8, colony formation assay, and 7-amino-actinomycin D/allophycocyanin double staining were performed to detect the function of miR-886-5p in MM cell lines. The expression of Bax and p53 was determined by western blot. RESULTS: The expression of miR-886-5p in the plasma of MM patients was higher than that in normal individuals and its level in MM cell lines was higher than that in peripheral blood mononuclear cells isolated from healthy individuals. miR-886-5p could trigger the cell proliferation and inhibition of apoptosis and affect the cell cycle. CONCLUSION: miR-886-5p triggered MM cell growth and may act as a diagnostic plasma biomarker for MM, potentially contributing to resistance to chemotherapy.


Assuntos
Biomarcadores Tumorais/sangue , Leucócitos Mononucleares/metabolismo , MicroRNAs/sangue , Mieloma Múltiplo/sangue , RNA Neoplásico/sangue , Adulto , Idoso , Apoptose , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Leucócitos Mononucleares/patologia , Masculino , Pessoa de Meia-Idade , Proteína Supressora de Tumor p53/biossíntese , Proteína X Associada a bcl-2/biossíntese
13.
Life Sci ; 232: 116583, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31226417

RESUMO

TP53 mutation is an indicator of poor prognostic in chronic lymphocytic leukemia (CLL). Worse still, CLL patients with TP53 mutation are associated with poor efficacy to current chemotherapeutic, such as Fludarabine. Here, we confirmed that high expression of HDAC1 in CLL patients with TP53 mutation, which is closely related to poor prognosis and drug-resistance. Subsequently, we demonstrated Entinostat (HDAC1 inhibitor) combination with Fludarabine significantly induced apoptosis in TP53 mutations CLL cells. Its mechanism was associated with up-regulation of the pro-apoptotic protein Bax and the down-regulation of HDAC1, HO-1 and BCL-2 proteins. More importantly, we also confirmed that upregulation of HDAC1 could resistant Entinostat-induced apoptosis in TP53 mutations CLL cells by activating the HDAC1/P38/HO-1 pathway. In vivo, we found that Entinostat combination with Fludarabine significantly induced tumor cells apoptosis and prolong survival time in xenograft mouse model. Finally, combining vitro and vivo experiments, we presented the first demonstration that Entinostat combination with Fludarabine had a synergistic effect on the induction of apoptosis in TP53 mutations CLL cells. In conclusion, we provide valuable pre-clinical experimental evidence for the treatment of CLL patients with poor prognosis, especially for TP53 mutations.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Benzamidas/farmacologia , Heme Oxigenase-1/metabolismo , Histona Desacetilase 1/metabolismo , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Piridinas/farmacologia , Proteína Supressora de Tumor p53/genética , Vidarabina/análogos & derivados , Idoso , Idoso de 80 Anos ou mais , Animais , Apoptose/efeitos dos fármacos , Benzamidas/administração & dosagem , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos , Sinergismo Farmacológico , Feminino , Histona Desacetilase 1/biossíntese , Histona Desacetilase 1/genética , Humanos , Leucemia Linfocítica Crônica de Células B/genética , Leucemia Linfocítica Crônica de Células B/metabolismo , Leucemia Linfocítica Crônica de Células B/patologia , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Piridinas/administração & dosagem , Distribuição Aleatória , Transdução de Sinais/efeitos dos fármacos , Vidarabina/administração & dosagem , Vidarabina/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto , Proteína X Associada a bcl-2/biossíntese , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
14.
Mol Cell Biochem ; 459(1-2): 131-139, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31114934

RESUMO

To investigate the expression status of FAM98A and its potential involvement in endometrial carcinoma, the relative expression of FAM98A in clinical endometrial carcinoma tissues was analyzed by immunohistochemistry and real-time polymerase chain reaction. Endogenous FAM98A protein was determined by Western blotting. The overall survival was calculated by the Kaplan-Meier's analysis. Cell growth/viability/proliferation was evaluated by cell counting, 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay, and clonogenic assay, respectively. Cell apoptosis was determined by the Annexin V/7-AAD double-staining methods followed by flow cytometry analysis. The regulatory effect of miR-142-3p on FAM98A was interrogated by luciferase reporter assay. Aberrant overexpression of FAM98A was found in endometrial carcinoma both in vitro and in vivo. Furthermore, high level of FMA98A was associated with poor prognosis. FAM98A deficiency in Ishikawa and RL95-2 cells significantly inhibited cell growth, cell viability, and cell proliferation. In addition, FAM98A-knockdown stimulated remarkable cell apoptosis, which might be mediated by down-regulation of BCL2 and up-regulation of BAX. Mechanistically, it was demonstrated that miR-142-3p directly targeted FAM98A, and modulated its expression. In conclusion, we unraveled the oncogenic properties of FAM98A in endometrial carcinoma and highlighted the miR-142-3p-FAM98A signaling in this disease.


Assuntos
Proliferação de Células , Neoplasias do Endométrio/metabolismo , Proteínas/metabolismo , Apoptose , Linhagem Celular Tumoral , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Feminino , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Proteínas/genética , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Proteína X Associada a bcl-2/biossíntese , Proteína X Associada a bcl-2/genética
15.
Environ Pollut ; 249: 482-490, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30928520

RESUMO

Epidemiological evidence showed that the particulate matter exposure is associated with atherosclerotic plaque progression, which may be related to foam cell formation, but the mechanism is still unknown. The study was aimed to investigate the toxic effects and possible mechanism of PM2.5 on the formation of macrophage foam cells induced by oxidized low density lipoprotein (ox-LDL). Results showed that PM2.5 induced cytotoxicity by decreasing the cell viability and increasing the LDH level in macrophage foam cells. PM2.5 aggravated the lipid accumulation in ox-LDL-stimulated macrophage RAW264.7 within markedly increasing level of intracellular lipid by Oil red O staining. The level of ROS increased obivously after co-exposure to PM2.5 and ox-LDL than single exposure group. In addition, serious mitochondrial damage such as the mitochondrial swelling, cristae rupturing and disappearance were observed in macrophage foam cells. The loss of the mitochondrial membrane potential (MMP) further exacerbated the mitochondrial damage in PM2.5-induced macrophage foam cells. The apoptotic rate increased more severely via up-regulated protein level of Bax, Cyt C, Caspase-9, Caspase-3, and down-regulated that of Bcl-2, indicating that PM2.5 activated the mitochondrial-mediated apoptosis pathway. In summary, our results demonstrated that PM2.5 aggravated the lipid accumulation, mitochondrial damage and apoptosis in macrophage foam cells, suggesting that PM2.5 was a risk factor of atherosclerosis progression.


Assuntos
Apoptose/efeitos dos fármacos , Células Espumosas/patologia , Lipoproteínas LDL/metabolismo , Potencial da Membrana Mitocondrial/fisiologia , Material Particulado/toxicidade , Placa Aterosclerótica/patologia , Animais , Aterosclerose/patologia , Caspase 3/biossíntese , Caspase 9/biossíntese , Linhagem Celular , Citocromos c/biossíntese , Metabolismo dos Lipídeos/efeitos dos fármacos , Camundongos , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima , Proteína X Associada a bcl-2/biossíntese
16.
Biomed Res Int ; 2019: 1434538, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30993110

RESUMO

Osteoarthritis (OA) is a chronic degenerative joint disease, where chondrocyte apoptosis is responsible for cartilage degeneration. Bax is a well-known proapoptotic protein of the Bcl-2 family, involved in a large number of physiological and pathological processes. However, the regulation mechanisms of Bax underlying chondrocyte apoptosis in OA remain unknown. In the present study, we determined the role of Bax in human OA and chondrocyte apoptosis. The results showed that Bax was upregulated in chondrocytes from the articular cartilage of OA patients and in cultured chondrocyte-like ATDC5 cells treated by IL-1ß. Bax was identified to be the direct target of miR-29a by luciferase reporter assay and by western blotting. Inhibition of miR-29a by the mimics protested and overexpression by miR-29a inhibitors aggravated ATDC5 apoptosis induced by IL-1ß. These data reveal that miR-29a/Bax axis plays an important role in regulating chondrocyte apoptosis and suggest that targeting the proapoptotic protein Bax and increasing expression levels of miR-29a emerge as potential approach for protection against the development of OA.


Assuntos
Apoptose , Condrócitos/metabolismo , Regulação da Expressão Gênica , MicroRNAs/biossíntese , Osteoartrite/metabolismo , Proteína X Associada a bcl-2/biossíntese , Idoso , Idoso de 80 Anos ou mais , Condrócitos/patologia , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Osteoartrite/genética , Osteoartrite/patologia , Proteína X Associada a bcl-2/genética
17.
J Biochem Mol Toxicol ; 33(7): e22324, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30951608

RESUMO

INTRODUCTION: Due to their unique properties including cellular uptake and the delivery efficiency to biological systems, nanoparticles are used in various preclinical and clinical applications. The aim of this study was to investigate the toxicity impacts of zinc oxide nanoparticles (ZnO-NPs) on morphology and functionality of the rat's liver and spleen and illustrated its safe-therapeutic doses. METHODS: The 28 female Swiss albino rats (180-220 g) and two human hepatocyte cell lines (HepG2 and HUH7) were designed as an in vivo and in vitro study, respectively. Samples were treated with certain doses of ZnO-NPs. The rat's liver morphology and functionality and apoptotic genes expression profile (Bax, Bcl-2, and P53) were analyzed to detect the cytotoxicity and antitumor impacts of ZnO-NPs, respectively. RESULTS: The results showed a positive significant association between the increasing doses of ZnO-NPs and alanine aminotransferase/aspartate aminotransferase values. Moreover, a meaningful correlation was detected between the rat's liver and spleen weight and ZnO-NPs doses. Furthermore, the histopathological analysis of rat's liver showed the individual cytotoxic properties of ZnO-NPs. Finally, the positive significant correlation was detected among the expression of Bax and P53 genes with ZnO-NPs. In addition, the negative correlation was demonstrated between the expression of Bcl-2 and ZnO-NPs. CONCLUSION: In general, in the current study, the antitumor effects of ZnO-NPs were confirmed by the enhancement of P53 and Bax genes expression profile, which are indicated the apoptotic induction in HUH7 cell line. Moreover, we introduced a safe-clinical ZnO-NPs dosage, have antitumor effects.


Assuntos
Citotoxinas , Neoplasias Hepáticas , Fígado , Nanopartículas , Baço , Óxido de Zinco , Animais , Citotoxinas/química , Citotoxinas/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Fígado/metabolismo , Fígado/patologia , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Nanopartículas/química , Nanopartículas/uso terapêutico , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Ratos , Baço/metabolismo , Baço/patologia , Proteína Supressora de Tumor p53/biossíntese , Óxido de Zinco/química , Óxido de Zinco/farmacologia , Proteína X Associada a bcl-2/biossíntese
18.
Protein Expr Purif ; 158: 20-26, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30738180

RESUMO

Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is crucial for the onset of apoptosis by perforating the mitochondrial outer membrane (MOM). This process can be seen as point of no return, since disintegration of the MOM leads to the release of apotogenic factors such as cytochrome c into the cytosol triggering the activation of caspases and subsequent apoptotic steps. Bax is able to interact with the MOM with both its termini, making it inherently difficult to express in E. coli. In this study, we present a novel approach to express and purify full-length Bax with significantly increased yields, when compared to the commonly applied strategy. Using a double fusion approach with an N-terminal GST-tag and a C-terminal Intein-CBD-tag, we were able to render both Bax termini inactive and prevent disruptive interactions from occurring during gene expression. By deploying an Intein-CBD-tag at the C-terminus we were further able to avoid the introduction of any artificial residues, hence ensuring the native like activity of the membrane-penetrating C-terminus of Bax. Further, by engineering a His6-tag to the C-terminus of the CBD-tag we greatly improved the robustness of the purification procedure. We report yields for pure, full-length Bax protein that are increased by an order of magnitude, when compared to commonly used Bax expression protocols.


Assuntos
Expressão Gênica , Proteínas Recombinantes de Fusão , Proteína X Associada a bcl-2 , Cristalografia por Raios X , Humanos , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteína X Associada a bcl-2/biossíntese , Proteína X Associada a bcl-2/química , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/isolamento & purificação
19.
J Steroid Biochem Mol Biol ; 185: 225-236, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30227242

RESUMO

Visfatin is an adipokine which has an endocrine effect on reproductive functions and regulates ovarian steroidogenesis. There is scant information about the expression, regulation, and functions of visfatin in the mammalian uterus. The present study examined expression and localization of visfatin in the mouse uterus at various stages of the natural estrous cycle, effects of estrogen and progesterone on localization and expression of visfatin in the ovariectomised mouse uterus and effect of visfatin inhibition by a specific inhibitor, FK866 on proliferation and apoptosis in the uterus. Western blot analysis of visfatin showed high expression in proestrus and metestrus while it declined in estrus and diestrus. Immulocalization study also showed strong immunostaining in the cells of endometrium, myometrium, luminal and glandular epithelium during proestrus and metestrus that estrus and diestrus. The uterine visfatin expression closely related to the increased estrogen levels in proestrus and suppressed when progesterone rose to a high level in diestrus. The treatment with estrogen to ovariectomised mice up-regulates visfatin, PCNA, and active caspase3 whereas progesterone up-regulates PCNA and down-regulates visfatin and active caspase3 expression in mouse uterus. The co-treatment with estrogen and progesterone up-regulates visfatin and down-regulates PCNA and active caspase3. In vitro study showed endogenous visfatin inhibition by FK866 increased expression of PCNA and BCL2 increased catalase activity while FK866 treatment decreased expression of active caspase3 and BAX with decreased SOD and GPx activity. BrdU labeling showed that inhibition of visfatin modulates the uterine proliferation. This study showed that expression of visfatin protein is steroid dependent in mouse uterus which is involved in the regulation of proliferation and apoptosis via modulating antioxidant system in the uterus of mice during the reproductive cycle.


Assuntos
Apoptose/fisiologia , Proliferação de Células/fisiologia , Endométrio/metabolismo , Estrogênios/metabolismo , Ciclo Estral/metabolismo , Miométrio/metabolismo , Nicotinamida Fosforribosiltransferase/biossíntese , Progesterona/metabolismo , Acrilamidas/farmacologia , Animais , Caspase 3/biossíntese , Catalase/biossíntese , Diestro/metabolismo , Estro/metabolismo , Feminino , Glutationa Peroxidase/biossíntese , Camundongos , Nicotinamida Fosforribosiltransferase/antagonistas & inibidores , Piperidinas/farmacologia , Proestro/metabolismo , Antígeno Nuclear de Célula em Proliferação/biossíntese , Superóxido Dismutase/biossíntese , Proteína X Associada a bcl-2/biossíntese
20.
Int J Mol Med ; 43(2): 693-700, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30483729

RESUMO

Oxidative stress serves an important role in hypertensive brain damage. Peroxisome proliferator­activated receptor γ (PPAR­Î³) agonists possess antioxidative and anti­apoptotic effects. The present study verified the possibility that rosiglitazone serves a neuroprotective role by alleviating oxidative stress and cell apoptosis in the hippocampi of spontaneously hypertensive rats (SHRs). SHRs and age­matched Wistar­Kyoto (WKY; both 56 weeks old) rats received gavage administration of vehicle or rosiglitazone (5 mg/kg/day) for eight weeks. Systolic blood pressure (SBP) was measured by the indirect tail­cuff method. The expression ratio of activated astrocytes was analyzed by glial fibrillary acidic protein immunohistochemistry. PPAR­Î³, inducible nitric oxide synthase (iNOS), gp47phox, B­cell lymphoma 2 (Bcl­2), Bcl­2­associated X protein (Bax) and caspase­3 expression were investigated by quantitative polymerase chain reaction and western blot analysis. The number of apoptotic cells in the hippocampus of four groups was detected using the terminal deoxynucleotidyl transferase­mediated dUTP end­labeling (TUNEL) method. Compared with the WKY group, the SHR group exhibited decreased Bcl­2 and PPAR­Î³ expression, increased SBP, increased ratio of activated astrocytes and TUNEL­positive cells, increased expression of iNOS, gp47phox, caspase­3 and Bax. Rosiglitazone administration increased Bcl­2 and PPAR­Î³ expression, decreased the ratio of activated astrocytes and TUNEL­positive cells, decreased iNOS, gp47phox, caspase­3 and Bax expression in the hippocampi of SHRs. However, rosiglitazone did not significantly decreased SBP in the SHR group. Therefore, rosiglitazone exerts neuroprotective effect through antioxidative and anti­apoptotic pathways, which was independent of blood pressure control.


Assuntos
Apoptose/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Hipertensão/fisiopatologia , Hipoglicemiantes/administração & dosagem , Rosiglitazona/administração & dosagem , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Astrócitos/patologia , Pressão Sanguínea/efeitos dos fármacos , Caspase 3/biossíntese , Hipocampo/metabolismo , Hipocampo/fisiopatologia , Hipertensão/metabolismo , Hipertensão/patologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Espécies Reativas de Oxigênio/metabolismo , Proteína X Associada a bcl-2/biossíntese , Proteína de Morte Celular Associada a bcl/biossíntese
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