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1.
Int J Mol Sci ; 20(15)2019 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-31390836

RESUMO

5-Lipoxygenase (5-LO) has been reported to be highly expressed in brain tumors and to promote glioma cell proliferation. Therefore, we investigated the anticancer activity of the novel 5-LO inhibitor derivative 3-tridecyl-4,5-dimethoxybenzene-1,2-diol hydroquinone (EA-100C red) on glioblastoma (GBM) cell growth. Cell viability was evaluated by MTT assay. The effects of the compound on apoptosis, oxidative stress and autophagy were assessed by flow cytometry (FACS). The mode of action was confirmed by Taqman apoptosis array, Real Time qPCR, confocal microscopy analysis and the western blotting technique. Our results showed that EA-100C Red had a higher anti-proliferative effect on LN229 as compared to U87MG cells. The compound induced a significant increase of apoptosis and autophagy and up-regulated pro-apoptotic genes (Bcl3, BNIP3L, and NFKBIA) in both GBM cell lines. In this light, we studied the effects of EA-100C red on the expression of CHOP and XBP1, that are implicated in ER-stress-mediated cell death. In summary, our findings revealed that EA-100C red induced ER stress-mediated apoptosis associated to autophagy in GBM cells through CHOP and Beclin1 up-regulation and activation of caspases 3, 9, JNK and NF-kappaB pathway. On these bases, EA-100C red could represent a promising compound for anti-cancer treatment.


Assuntos
Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Neoplasias Encefálicas/metabolismo , Glioblastoma/metabolismo , Hidroquinonas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Resposta a Proteínas não Dobradas/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Estrutura Molecular , Transdução de Sinais/efeitos dos fármacos
2.
Appl Biochem Biotechnol ; 189(4): 1291-1303, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31254228

RESUMO

With growing concern about the possible risks and side effects of antibiotic drugs, more and more natural products with antibacterial activity are studied as the substitutes. In this paper, the antibacterial activity of hydroquinone and arbutin in Ainsliaea bonatii was investigated, which both displayed relatively strong antibacterial activity against Staphylococcus aureus (SA), methicillin-resistant S. aureus (MRSA), and extended spectrum ß-lactamase S. aureus (ESBL-SA). The antibacterial mechanism of hydroquinone had been explored by scanning electron microscopy (SEM), alkaline phosphatase (AKP), and bacterial extracellular protein leakage. Results showed that hydroquinone could destroy the bacterial cell wall and membrane, increase permeability, lead leakage of intracellular substance affect synthesis of protein, and influence expression of genes.


Assuntos
Antibacterianos/farmacologia , Arbutina/farmacologia , Hidroquinonas/farmacologia , Lamiaceae/química , Staphylococcus aureus Resistente à Meticilina/metabolismo , Antibacterianos/química , Arbutina/química , Hidroquinonas/química , Staphylococcus aureus Resistente à Meticilina/ultraestrutura
3.
Oxid Med Cell Longev ; 2019: 3501059, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31089407

RESUMO

Dental resin monomers such as 2-hydroxyethyl methacrylate (HEMA) disturb vital cell functions and induce mitochondrial intrinsic apoptosis via generation of oxidative stress. Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates the gene expression of antioxidative enzymes and plays a crucial role in the maintenance of cellular redox equilibrium and mitochondrial homeostasis. The present study investigated the functional significance of Nrf2 in cellular response toward HEMA. It was found that HEMA stimulation promoted nuclear translocation of Nrf2 and increased Nrf2 and heme oxygenase-1 (HO-1) expression, which was further enhanced by Nrf2 activator tert-butylhydroquinone (tBHQ), but suppressed by Nrf2 inhibitor ML385. Pretreatment of primary human dental pulp cells (hDPCs) with tBHQ protected the cells from HEMA-induced oxidative injury (increased reactive oxygen species production and apoptosis) and mitochondrial impairment (morphological alterations, decreased ATP production, suppressed oxidative phosphorylation activity, depolarization of mitochondrial membrane potential, and disrupted electron transport chain). In contrast, pretreatment with ML385 increased cell sensitivity to these injurious processes. This protective effect on mitochondria could be related to peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α)/nuclear respiratory factor 1 (NRF1) pathway. These results contribute to the understanding of the function of Nrf2 and the development of novel therapies to counteract the adverse effects of dental resin monomers.


Assuntos
Metacrilatos/toxicidade , Mitocôndrias/patologia , Fator 2 Relacionado a NF-E2/metabolismo , Substâncias Protetoras/metabolismo , Adolescente , Adulto , Apoptose/efeitos dos fármacos , Polpa Dentária/patologia , Polpa Dentária/ultraestrutura , Heme Oxigenase-1/metabolismo , Humanos , Hidroquinonas/farmacologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/ultraestrutura , Modelos Biológicos , Fator 1 Nuclear Respiratório/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Adulto Jovem
4.
Int J Mol Sci ; 20(7)2019 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-30959808

RESUMO

: Carnosic acid (CA) is a phytochemical found in some dietary herbs, such as Rosmarinus officinalis L., and possesses antioxidative and anti-microbial properties. We previously demonstrated that CA functions as an activator of nuclear factor, erythroid 2 (NF-E2)-related factor 2 (Nrf2), an oxidative stress-responsive transcription factor in human and rodent cells. CA enhances the expression of nerve growth factor (NGF) and antioxidant genes, such as HO-1 in an Nrf2-dependent manner in U373MG human astrocytoma cells. However, CA also induces NGF gene expression in an Nrf2-independent manner, since 50 µM of CA administration showed striking NGF gene induction compared with the classical Nrf2 inducer tert-butylhydroquinone (tBHQ) in U373MG cells. By comparative transcriptome analysis, we found that CA activates activating transcription factor 4 (ATF4) in addition to Nrf2 at high doses. CA activated ATF4 in phospho-eIF2α- and heme-regulated inhibitor kinase (HRI)-dependent manners, indicating that CA activates ATF4 through the integrated stress response (ISR) pathway. Furthermore, CA activated Nrf2 and ATF4 cooperatively enhanced the expression of NGF and many antioxidant genes while acting independently to certain client genes. Taken together, these results represent a novel mechanism of CA-mediated gene regulation evoked by Nrf2 and ATF4 cooperation.


Assuntos
/farmacologia , Fator 4 Ativador da Transcrição/genética , Citoproteção/genética , Regulação da Expressão Gênica , Fator 2 Relacionado a NF-E2/genética , Fator 4 Ativador da Transcrição/metabolismo , Aldeído Redutase/genética , Aldeído Redutase/metabolismo , Aldo-Ceto Redutases , Antioxidantes/metabolismo , Linhagem Celular Tumoral , Citoproteção/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Hidroquinonas/farmacologia , Modelos Biológicos , Fator 2 Relacionado a NF-E2/metabolismo , Fator de Crescimento Neural/genética , Fator de Crescimento Neural/metabolismo , Transdução de Sinais/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos , Tunicamicina/farmacologia
5.
Molecules ; 24(5)2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30823626

RESUMO

1-O-Hexyl-2,3,5-trimethylhydroquinone (HTHQ) has previously been found to have effective anti-oxidant and anti-lipid-peroxidative activity. We aimed to elucidate whether HTHQ can prevent dopaminergic neuronal cell death by investigating the effect on l-DOPA-induced cytotoxicity in PC12 cells. HTHQ protected from both l-DOPA-induced cell death and superoxide dismutase activity reduction. When assessing the effect of HTHQ on oxidative stress-related signaling pathways, HTHQ inhibited l-DOPA-induced phosphorylation of sustained extracellular signal-regulated kinases (ERK1/2), p38 mitogen-activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK1/2). HTHQ also normalized l-DOPA-reduced Bcl-2-associated death protein (Bad) phosphorylation and Bcl-2-associated X protein (Bax) expression, promoting cell survival. Taken together, HTHQ exhibits protective effects against l-DOPA-induced cell death through modulation of the ERK1/2-p38MAPK-JNK1/2-Bad-Bax signaling pathway in PC12 cells. These results suggest that HTHQ may show ameliorative effects against oxidative stress-induced dopaminergic neuronal cell death, although further studies in animal models of Parkinson's disease are required to confirm this.


Assuntos
Neurônios Dopaminérgicos/efeitos dos fármacos , Hidroquinonas/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Doença de Parkinson/tratamento farmacológico , Animais , Morte Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neurônios Dopaminérgicos/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/genética , Levodopa/efeitos adversos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Células PC12 , Doença de Parkinson/genética , Doença de Parkinson/patologia , Ratos , Proteína X Associada a bcl-2/genética , Proteína de Morte Celular Associada a bcl/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética
6.
Biochem Pharmacol ; 163: 321-334, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30825431

RESUMO

The endocannabinoid system (ECS) may play a role in the pathophysiology of systemic sclerosis (SSc). Cannabinoids acting as dual PPARγ/CB2 agonists, such as VCE-004.8 and Ajulemic acid (AjA), have been shown to alleviate skin fibrosis and inflammation in SSc models. Since both compounds are being tested in humans, we compared their activities in the bleomycin (BLM) SSc model. Specifically, the pharmacotranscriptomic signature of the compounds was determined by RNA-Seq changes in the skin of BLM mice treated orally with AjA or EHP-101, a lipidic formulation of VCE-004.8. While both compounds down-regulated the expression of genes involved in the inflammatory and fibrotic components of the disease and the pharmacotranscriptomic signatures were similar for both compounds in some pathways, we found key differences between the compounds in vasculogenesis. Additionally, we found 28 specific genes with translation potential by comparing with a list of human scleroderma genes. Immunohistochemical analysis revealed that both compounds prevented fibrosis, collagen accumulation and Tenascin C (TNC) expression. The endothelial CD31+/CD34+ cells and telocytes were reduced in BLM mice and restored only by EHP-101 treatment. Finally, differences were found in plasmatic biomarker analysis; EHP-101, but not AjA, enhanced the expression of some factors related to angiogenesis and vasculogenesis. Altogether the results indicate that dual PPARγ/CB2 agonists qualify as a novel therapeutic approach for the treatment of SSc and other fibrotic diseases. EHP-101 demonstrated unique mechanisms of action related to the pathophysiology of SSc that could be beneficial in the treatment of this complex disease without current therapeutic options.


Assuntos
Canabinoides/farmacologia , Dronabinol/análogos & derivados , Regulação da Expressão Gênica/efeitos dos fármacos , Hidroquinonas/farmacologia , PPAR gama/agonistas , Receptor CB2 de Canabinoide/agonistas , Escleroderma Sistêmico/tratamento farmacológico , Animais , Biomarcadores , Bleomicina/toxicidade , Dronabinol/administração & dosagem , Dronabinol/farmacologia , Feminino , Fibrose/induzido quimicamente , Hidroquinonas/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , PPAR gama/genética , PPAR gama/metabolismo , Fibrose Pulmonar , Receptor CB2 de Canabinoide/genética , Receptor CB2 de Canabinoide/metabolismo , Pele/efeitos dos fármacos , Pele/patologia
7.
Biochem Biophys Res Commun ; 511(3): 637-643, 2019 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-30826055

RESUMO

Metabolic bone diseases are global public health concerns and are primarily caused by uncontrolled osteoclast (OC) formation and activation. During OC differentiation, intracellular reactive oxygen species (ROS) stimulated by receptor activator of nuclear factor kappa-B ligand (RANKL) can serve as the signaling molecules to promote osteoclastic genes expression. Nuclear factor erythroid-2 related factor 2 (NRF2), a master mediator of cellular antioxidant response, also plays a critical role in OC differentiation through the regulation of redox homeostasis. In this study, we investigated the effects of three NRF2 inducers on osteoclastogenesis, including Bardoxolone methyl (CDDO-Me), Sulforaphane (SFN), and tert-butylhydroquinone (tBHQ). By treating RAW cells with three compounds, we found that NRF2 was activated and its downstream antioxidant genes were upregulated, and the RANKL-induced intracellular ROS production and osteoclastogenesis were impaired. Additionally, the expression of nuclear factor of activated T cells c1 (NFATC1), C-FOS and tumor necrosis factor alpha (TNFα) were inhibited after acute exposures (6 h) to the three compounds. Furthermore, suppressed the expression of osteoclast differentiation-associated genes, tartrate-resistant acid phosphatase (TRAP), cathepsin K (CTSK), matrix metalloproteinase-9 (MMP-9) and dendritic cell-specific transmembrane protein (DC-STAMP) were observed after prolonged exposures (5 days) to the compounds. Taken together, these results suggest that CDDO-Me, SFN and tBHQ attenuate RANKL-induced osteoclastogenesis via activation of NRF2-mediated antioxidant response. Among these compounds, relatively low concentrations of CDDO-Me showed stronger active and inhibitory effects on antioxidant response and osteoclastogenesis, respectively.


Assuntos
Antioxidantes/farmacologia , Hidroquinonas/farmacologia , Isotiocianatos/farmacologia , Ácido Oleanólico/análogos & derivados , Osteogênese/efeitos dos fármacos , Ligante RANK/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Masculino , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/metabolismo , Ácido Oleanólico/farmacologia , Osteoclastos/citologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Espécies Reativas de Oxigênio/metabolismo
8.
Chem Biol Interact ; 302: 1-10, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30703377

RESUMO

2,3,5-tris(Glutathion-S-yl)hydroquinone (TGHQ), a potent nephrotoxic and nephroncarcinogenic metabolite of benzene and hydroquinone, retains the ability to redox cycle and create oxidative stress. We have previously detected that TGHQ induces ROS-dependent necrotic or apoptotic cell death in renal epithelial HK-2 and human leukemic HL-60 cells respectively. Herein, we sought to determine the nature of the Nrf2 regulation in HK-2 and HL-60 cells undergoing TGHQ-mediated ROS-dependent cell death, due to the key role of Nrf2 in oxidative stress. Intriguingly, Nrf2 was upregulated in HK-2, but not in HL-60 cells, despite the ROS-dependent nature of cell death in both cell types. The possibility that TGHQ targeted the GSK3ß-dependent Nrf2 stabilization pathway in HL-60 cells was discounted, whereas TGHQ-induced decreases in Nrf2 phosphorylation at Ser40 site appears to partially underlie the inability of TGHQ to up-regulate Nrf2 expression in HL-60 cells. Moreover, whereas the TGHQ-induced post-translational stabilization of Nrf2 in HK-2 cells resulted in the expected upregulation of HO1 and NQO1 mRNA, TGHQ actually decreased Nrf2 mRNA in HL-60 cells, with a concomitant decrease in NQO1, but not HO1 mRNA. In summary, we define differences between the two cell types that might contribute to the engagement of the Nrf2 signaling pathways. By extension, these data provide evidence that Nrf2 is not necessarily activated in ROS-dependent cell death, and further delve into the knowledge that Nrf2 regulation sensing by cells might be achieved at solely transcriptional level, not related to its degradation.


Assuntos
Apoptose/efeitos dos fármacos , Glutationa/análogos & derivados , Hidroquinonas/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Linhagem Celular , Glutationa/química , Glutationa/farmacologia , Glicogênio Sintase Quinase 3 beta/metabolismo , Células HL-60 , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Humanos , Peróxido de Hidrogênio/farmacologia , Hidroquinonas/química , Leupeptinas/farmacologia , NAD(P)H Desidrogenase (Quinona)/genética , NAD(P)H Desidrogenase (Quinona)/metabolismo , Fosforilação/efeitos dos fármacos , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteína Quinase C/metabolismo , Regulação para Cima/efeitos dos fármacos
9.
Exp Parasitol ; 199: 17-23, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30790574

RESUMO

Exposing second-stage juveniles (J2) of Meloidogyne incognita in vitro to a phenolic compound sometimes fails to cause J2 mortality, but in tests in vivo the same compound may reduce the infectivity and population of the nematode. This work aimed to study the effect of phenolic compounds on M. incognita through in vitro and in vivo assays. In the in vitro assay 49 phenolic compounds were screened for their toxicity to M. incognita J2. As a result, D-(-)-4-hydroxyphenylglycine, t-butylhydroquinone, L-3-(3,4-dihydroxyphenyl)alanine, sesamol, 2,4-dihydroxyacetophenone, and p-anisaldehyde increased the J2 mortality. These compounds presented, respectively, the following lethal concentrations to 50% of J2 (LC50): 365, 352, 251, 218, 210, and 85 µg/mL, while Carbofuran (positive control) had 150 µg/mL. However, none of these compounds were efficient in controlling the nematode in inoculated tomato plants, even when 2.77-fold of their LC50 were used. Although inactive in the in vitro test at 500 µg/mL, hydroquinone (3.5 mg per plant) reduced M. incognita population and galls by up to 99% to levels similar to the nematicide Carbofuran (1.2 mg per plant). Additionally, hydroquinone increased the root weight when compared to the negative and positive controls, water/NaOH and Carbofuran, respectively. In this study, we showed that some phenolic compounds, hydroquinone in particular, revealed a potential new option for the control of M. incognita.


Assuntos
Lycopersicon esculentum/parasitologia , Fenóis/farmacologia , Tylenchoidea/efeitos dos fármacos , Animais , Arbutina/administração & dosagem , Arbutina/química , Arbutina/farmacologia , Benzaldeídos/administração & dosagem , Benzaldeídos/química , Benzaldeídos/farmacologia , Ácidos Cafeicos/administração & dosagem , Ácidos Cafeicos/química , Ácidos Cafeicos/farmacologia , Carbofurano/administração & dosagem , Carbofurano/química , Carbofurano/farmacologia , Catecóis/administração & dosagem , Catecóis/química , Catecóis/farmacologia , Glicerol/administração & dosagem , Glicerol/química , Glicerol/farmacologia , Hidroquinonas/administração & dosagem , Hidroquinonas/química , Hidroquinonas/farmacologia , Hidroxibenzoatos/administração & dosagem , Hidroxibenzoatos/química , Hidroxibenzoatos/farmacologia , Dose Letal Mediana , Naftóis/administração & dosagem , Naftóis/química , Naftóis/farmacologia , Fenóis/administração & dosagem , Fenóis/química , Distribuição Aleatória , Resorcinóis/administração & dosagem , Resorcinóis/química , Resorcinóis/farmacologia , Fatores de Tempo
10.
Phytomedicine ; 55: 1-8, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30668419

RESUMO

BACKGROUND: Many bone-related diseases such as osteoporosis and rheumatoid arthritis are commonly associated with excessive activity of the osteoclast. Ganomycin I (GMI), a meroterpenoid isolated from Vietnamese mushroom Ganoderma lucidum, possesses a variety of beneficial effects on human health. However, its impact and underlying mechanism on osteoclastogenesis remain unclear. In the present study, we investigated the effect of GMI on RANKL-induced osteoclast formation in mouse BMMs and RAW264.7 cells. METHODS: BMMs or RAW264.7 cells were treated with GMI followed by an evaluation of cell viability, RANKL-induced osteoclast differentiation, actin-ring formation, and resorption pits activity. Effects of GMI on RANKL-induced phosphorylation of MAPKs as well as the expression levels of NFATc1 and c-Fos were evaluated by Western blot analysis. Expression levels of osteoclast marker genes were evaluated by Western blot analysis and reverse transcription-qPCR. RESULTS: GMI significantly inhibited RANKL-induced osteoclast differentiation by decreasing the number of osteoclasts, osteoclast actin-ring formation, and bone resorption in a dose-dependent manner without affecting cell viability. At molecular level, GMI inhibited the RANKL-induced phosphorylation of ERK, JNK, and p38 MAPKs, as well as the expression levels of c-Fos and NFATc1, which are known to be crucial transcription factors for osteoclast formation. In addition, GMI decreased expression levels of osteoclastogenesis specific marker genes including c-Src, CtsK, TRAP, MMP-9, OSCAR, and DC-STAMP in RANKL-stimulated BMMs. CONCLUSION: Our findings suggest that GMI can attenuate osteoclast formation by suppressing RANKL-mediated MAPKs and NFATc1 signaling pathways and the anti-osteoclastogenic activity of GMI may extend our understanding of molecular mechanisms underlying biological activities and pharmacological use of G. lucidum as a traditional anti-osteoporotic medicine.


Assuntos
MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , Hidroquinonas/farmacologia , Fatores de Transcrição NFATC/antagonistas & inibidores , Osteogênese/efeitos dos fármacos , Ligante RANK/metabolismo , Animais , Reabsorção Óssea/tratamento farmacológico , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica/efeitos dos fármacos , Hidroquinonas/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos ICR , Fatores de Transcrição NFATC/metabolismo , Osteoclastos/citologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/fisiologia , Osteogênese/fisiologia , Fosforilação/efeitos dos fármacos , Células RAW 264.7 , Reishi/química
11.
Int Ophthalmol ; 39(10): 2303-2312, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30637543

RESUMO

OBJECTIVE: The objective of this work was to find out the effects of nuclear factor erythroid 2-related factor/heme oxygenase-1 (Nrf2/HO-1) pathway on retinal ganglion cell (RGC) injury in glaucoma. METHODS: The chronic ocular hypertension (COH) rat models of glaucoma were constructed, and intraocular pressure (IOP) and RGC numbers were detected at different time points. Additionally, rats were divided into normal group (normal control rats), model group (COH model rats), and model + tBHQ group (COH model rats treated with Nrf activator, tBHQ). RGC apoptosis was detected by using TUNEL staining, and the expressions of Nrf2/HO-1 were detected by qRT-PCR and western blotting. RESULTS: COH model rats showed significant IOP elevation and the increased mRNA and protein expressions of Nrf2 and HO-1 from 1 to 6 weeks after operation, with the evidently decreased RGC numbers at 4 weeks and 6 weeks after operation (all P < 0.05). Besides, rats in the model group had increased apoptosis index (AI) of RGCs and the elevated mRNA and protein expressions of Nrf2/HO-1 with remarkably reduced RGC numbers when compared with normal control rats, but the model rats treated with tBHQ exhibited an apparent decrease in AI of RGCs, as well as remarkable increases in RGC numbers and the mRNA and protein expression of Nrf2/HO-1 (all P < 0.05). CONCLUSION: Activation of Nrf2/HO-1 pathway significantly reduced the apoptosis and injury of RGCs in rats with chronic ocular hypertension (COH), thereby protecting RGCs in glaucoma, which could be a promising clinical target to prevent RGC degeneration in glaucoma.


Assuntos
Glaucoma/fisiopatologia , Heme Oxigenase-1/fisiologia , Fator 2 Relacionado a NF-E2/fisiologia , Hipertensão Ocular/fisiopatologia , Células Ganglionares da Retina/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Doença Crônica , Modelos Animais de Doenças , Hidroquinonas/farmacologia , Ratos
12.
Toxicol Appl Pharmacol ; 363: 64-71, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30476502

RESUMO

Lead (Pb) is one of the most widely studied occupational and environmental toxins. Chronic exposure to Pb affects neural function in the central nervous system (CNS). Glial cells in the CNS, such as microglia and astrocytes, respond differently to Pb-induced toxicity. However, the underlying mechanism has not yet been identified. We measured the cell viability and intracellular Pb uptake in rat primary microglia and astrocytes using the CCK-8 assay and inductively coupled plasma mass spectrometry, and found that Pb decreased microglial viability at lower dosages than in astrocytes, while Pb uptake was greater in astrocytes. Pb-induced oxidative stress in microglia results in increased production of reactive oxygen species, down-regulation of glutathione, and enhanced Nrf2 protein expression, while there was no obvious change in astrocytes. The role of Nrf2 in Pb-induced oxidative stress has also been confirmed in primary microglia with the use of Nrf2 small interfering RNA and an Nrf2 agonist. These data indicate that primary microglia were more sensitive to Pb exposure than astrocytes, which is associated with an obvious oxidative stress response and up-regulation of Nrf2 might be involved in this process.


Assuntos
Astrócitos/efeitos dos fármacos , Chumbo/toxicidade , Microglia/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Animais , Animais Recém-Nascidos , Astrócitos/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Regulação para Baixo/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Glutationa/metabolismo , Hidroquinonas/farmacologia , Microglia/metabolismo , Fator 2 Relacionado a NF-E2/agonistas , Fator 2 Relacionado a NF-E2/genética , Estresse Oxidativo/efeitos dos fármacos , Cultura Primária de Células , RNA Interferente Pequeno/metabolismo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Testes de Toxicidade Aguda , Testes de Toxicidade Crônica , Regulação para Cima/efeitos dos fármacos
13.
Methods Mol Biol ; 1894: 229-246, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30547464

RESUMO

Transcription factor Nrf2, nuclear factor (erythroid-derived 2)-like 2, is considered a master regulator of redox homeostasis and plays a central role in antioxidant and anti-inflammatory defence. It has been largely reported that oxidative stress is implicated in nanoparticle-induced toxicity with the involvement of Nrf2. Several basic methods for Nrf2 evaluation with exposure to nanoparticles are described in this chapter including real-time reverse transcription-polymerase chain reaction (RT-PCR), western blotting, immunofluorescence staining, electrophoretic mobility shift assay, DNase I footprinting, dimethylsulfate footprinting, protein pulse-chase analysis, and tert-butylhydroquinone treatment.


Assuntos
Fator 2 Relacionado a NF-E2/análise , Nanopartículas/toxicidade , Animais , Western Blotting/instrumentação , Western Blotting/métodos , Células Cultivadas , Cicloeximida/farmacologia , Pegada de DNA/instrumentação , Pegada de DNA/métodos , Ensaio de Desvio de Mobilidade Eletroforética/instrumentação , Ensaio de Desvio de Mobilidade Eletroforética/métodos , Hidroquinonas/farmacologia , Microscopia Confocal , Fator 2 Relacionado a NF-E2/antagonistas & inibidores , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , RNA Mensageiro/isolamento & purificação , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/instrumentação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos
14.
Chem Res Toxicol ; 32(3): 421-436, 2019 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-30547568

RESUMO

Here we report a vertically integrated in vitro - in silico study that aims to elucidate the molecular initiating events involved in the induction of oxidative stress (OS) by seven diverse chemicals (cumene hydroperoxide, t-butyl hydroperoxide, hydroquinone, t-butyl hydroquinone, bisphenol A, Dinoseb, and perfluorooctanoic acid). To that end, we probe the relationship between chemical properties, cell viability, glutathione (GSH) depletion, and antioxidant gene expression. Concentration-dependent effects on cell viability were assessed by MTT assay in two Hepa-1 derived mouse liver cell lines: a control plasmid vector transfected cell line (Hepa-V), and a cell line with increased glutamate-cysteine ligase (GCL) activity and GSH content (CR17). Changes to intracellular GSH content and mRNA expression levels for the Nrf2-driven antioxidant genes Gclc, Gclm, heme oxygenase-1 ( Hmox1), and NADPH quinone oxidoreductase-1 ( Nqo1) were monitored after sublethal exposure to the chemicals. In silico models of covalent and redox reactivity were used to rationalize differences in activity of quinones and peroxides. Our findings show CR17 cells were generally more resistant to chemical toxicity and showed markedly attenuated induction of OS biomarkers; however, differences in viability effects between the two cell lines were not the same for all chemicals. The results highlight the vital role of GSH in protecting against oxidative stress-inducing chemicals as well as the importance of probing molecular initiating events in order to identify chemicals with lower potential to cause oxidative stress.


Assuntos
Antioxidantes/metabolismo , Expressão Gênica/efeitos dos fármacos , Glutationa/biossíntese , Glutationa/metabolismo , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , 2,4-Dinitrofenol/análogos & derivados , 2,4-Dinitrofenol/química , 2,4-Dinitrofenol/farmacologia , Animais , Derivados de Benzeno/química , Derivados de Benzeno/farmacologia , Compostos Benzidrílicos/química , Compostos Benzidrílicos/farmacologia , Caprilatos/química , Caprilatos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Fluorcarbonetos/química , Fluorcarbonetos/farmacologia , Hidroquinonas/química , Hidroquinonas/farmacologia , Cinética , Camundongos , Estrutura Molecular , Estresse Oxidativo/efeitos dos fármacos , Fenóis/química , Fenóis/farmacologia , terc-Butil Hidroperóxido/química , terc-Butil Hidroperóxido/farmacologia
15.
Bioorg Med Chem ; 27(1): 92-99, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30473361

RESUMO

BACKGROUND: The Nrf2-Keap1 interaction is the major regulatory pathway for cytoprotective responses against oxidative and electrophilic stresses. Keap1, a substrate protein of a Cul3-dependent E3 ubiquitin ligase complex, is a negative regulator of Nrf2. The use of chemicals to regulate the interaction between Keap1 and Nrf2 has been proposed as a strategy for the chemoprevention of degenerative diseases and cancers. RESULTS: The interactions between Keap1 and Nrf2 in vitro and in vivo were investigated using fluorescence resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC) strategies in our study. Nrf2 with its N-terminal fused to eGFP and Keap1 with its C-terminal fused to mCherry were expressed and purified in vitro. When purified eGFP-Nrf2 and Keap1-mChrry proteins were mixed together, a strong FRET signal could be detected, indicating an efficient energy transfer from eGFP to mCherry. Moreover, the FRET was detected in vivo using confocal microscopy in colon cancer HCT-116 cells that were co-transfected with eGFP-Nrf2 and Keap1-mCherry. Finally, using an eGFP BiFC approach, the Keap1-Nrf2 interaction was also detected in MCF7 cells by transfecting eGFP N-terminal fused to Nrf2 (eN158-Nrf2) and eGFP C-terminal fused to Keap1 (eC159-Keap1). Using the BiFC and FRET systems, we demonstrated that the prototypical Nrf2-activiting compound tBHQ and the antitumor drug F-dUrd might interfere with the intracellular interaction between Keap1 and Nrf2 whereas the 5-Fu have little role in activating the protective response of Nrf2 pathway in cancer cells. CONCLUSIONS: By analyzing the perturbation of the energy transfer between the donor and acceptor fluorophores and the bimolecular fluorescence complementation of eGFP, we can screen potential inhibitors for the interaction between Keap1 and Nrf2.


Assuntos
Floxuridina/farmacologia , Hidroquinonas/farmacologia , Proteína 1 Associada a ECH Semelhante a Kelch/antagonistas & inibidores , Fator 2 Relacionado a NF-E2/antagonistas & inibidores , Ligação Proteica/efeitos dos fármacos , Antimetabólitos Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Avaliação Pré-Clínica de Medicamentos , Transferência Ressonante de Energia de Fluorescência , Fluoruracila/farmacologia , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
16.
J Biochem Mol Toxicol ; 33(4): e22269, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30506647

RESUMO

Hydroquinone (HQ), one of the most significant metabolic activation products of benzene in an organism, can cause hematological toxicity, such as acute myeloid leukemia. It is a clear carcinogen that can cause changes in the disorder of cell cycle and cell growth. However, its molecular mechanisms remain unclear. E4 transcription factor 1 (E4F1), an important transcription factor, participating in the regulation of cell cycle may be related to the occurrence of tumor. Here, we examined the HQ-induced malignant transformed TK6 cells (TK6-HT) to illustrate the role of E4F1 in carcinogenesis. The present study showed that both the expressions of E4F1 messenger RNA and protein increased obviously in TK6-HT, preliminarily indicating that E4F1 is associated with HQ-induced carcinogenesis. To further explore the role of E4F1, we established E4F1 silencing TK6-HT (pLVX-shE4F1) and its control cells (pLVX-shNC) using lentiviral short hairpin RNA (shRNA) interference expression plasmid vector pLVX-shRNA. Flow cytometry and cell counting kit-8 assay were used to determine the effects of E4F1 silencing on cell cycle and cell growth, respectively. E4F1 silencing inhibited cell growth in TK6-HT. The results from flow cytometry indicated that the inhibitory effect on cell growth may be the results of the E4F1 silencing-induced accumulation in G2/M compared with TK6-HT-shNC. Meanwhile, levels of DNA damage (γ-H2AX), proteins of Rb and phosphorylated Rb, and reactive oxygen species were increased in TK6-HT-shRNA2 cells, which is the critical reason of cell-cycle arrest. In conclusion, E4F1 silencing inhibits the cell growth through cell-cycle arrest in malignant transformed cells induced by HQ.


Assuntos
Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/genética , Divisão Celular/efeitos dos fármacos , Divisão Celular/genética , Inativação Gênica , Hidroquinonas/farmacologia , Proteínas Repressoras/fisiologia , Linhagem Celular , Transformação Celular Neoplásica , Citometria de Fluxo , Histonas/metabolismo , Humanos , Espécies Reativas de Oxigênio/metabolismo , Proteínas Repressoras/genética
17.
Nat Prod Res ; 33(12): 1691-1698, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29382220

RESUMO

One new furylhydroquinone derivative (1) and seven known compounds (2-8) were isolated from the roots of Lithospermum erythrorhizon Sieb. et Zucc (Boraginaceae). The structure of 1 was elucidated by extensive spectroscopic methods using NMR and MS. The absolute configuration of shikonofuran J (1) was unambiguously determined by aid of comparison experimental ECD with predicted ECD spectra. All the isolates were tested for their inhibitory activities against IL-6 production in HaCaT cells stimulated by tumor necrosis factor (TNF)-α. It was found that gracicleistanthoside (5) and uridine (7) remarkably down-regulated the TNF-α-induced synthesis of interleukin-6 (IL-6), a pro-inflammatory cytokine associated with cutaneous inflammation, in HaCaT cells.


Assuntos
Anti-Inflamatórios/farmacologia , Hidroquinonas/isolamento & purificação , Lithospermum/química , Anti-Inflamatórios/isolamento & purificação , Linhagem Celular , Regulação para Baixo/efeitos dos fármacos , Humanos , Hidroquinonas/farmacologia , Interleucina-6/biossíntese , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Estrutura Molecular , Raízes de Plantas/química , Fator de Necrose Tumoral alfa/farmacologia
18.
Eur J Nutr ; 58(3): 989-996, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29330659

RESUMO

PURPOSE: The purpose of this study was to evaluate acute effects of coffee with a high content of chlorogenic acids and different hydroxyhydroquinone contents on postprandial endothelial dysfunction. METHODS: This was a single-blind, randomized, placebo-controlled, crossover-within-subject clinical trial. A total of 37 patients with borderline or stage 1 hypertension were randomized to two study groups. The participants consumed a test meal with a single intake of the test coffee. Subjects in the Study 1 group were randomized to single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone or coffee with a high content of chlorogenic acids and a high content of hydroxyhydroquinone with crossover. Subjects in the Study 2 group were randomized to single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone or placebo coffee with crossover. Endothelial function assessed by flow-mediated vasodilation and plasma concentration of 8-isoprostanes were measured at baseline and at 1 and 2 h after coffee intake. RESULTS: Compared with baseline values, single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone, but not coffee with a high content of chlorogenic acids and high content of hydroxyhydroquinone or placebo coffee, significantly improved postprandial flow-mediated vasodilation and decreased circulating 8-isoprostane levels. CONCLUSIONS: These findings suggest that a single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone is effective for improving postprandial endothelial dysfunction. CLINICAL TRIAL REGISTRATION: URL for Clinical Trial: https://upload.umin.ac.jp ; Registration Number for Clinical Trial: UMIN000013283.


Assuntos
Ácido Clorogênico/farmacologia , Café , Endotélio Vascular/efeitos dos fármacos , Hidroquinonas/farmacologia , Hipertensão/dietoterapia , Estudos Cross-Over , Endotélio Vascular/fisiopatologia , Feminino , Humanos , Hipertensão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Período Pós-Prandial , Método Simples-Cego
19.
J Nat Prod ; 81(11): 2364-2370, 2018 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-30375869

RESUMO

The first total synthesis of violaceoid A, a cytotoxic agent, and the asymmetric total synthesis of (-)- and (+)-violaceoid B are reported. The precursor was accessed by desymmetrization of a substituted quinol moiety, and the racemic secondary alcohol was kinetically resolved using a chiral nucleophilic catalyst. The asymmetric synthesis of (-)- and (+)-violaceoid B elucidated the absolute configuration of the naturally occurring violaceoid B. Synthetic violaceoid A inhibited the growth of human breast cancer cell lines MCF-7 and Hs 578T at concentrations of less than 100 µM, while ( S)- and ( R)-violaceoid B were inactive.


Assuntos
Hidroquinonas/síntese química , Catálise , Linhagem Celular Tumoral , Humanos , Hidroquinonas/química , Hidroquinonas/farmacologia , Estereoisomerismo
20.
Sci Rep ; 8(1): 13670, 2018 09 12.
Artigo em Inglês | MEDLINE | ID: mdl-30209334

RESUMO

Innate immune cells quickly infiltrate the site of pathogen entry and not only stave off infection but also initiate antigen presentation and promote adaptive immunity. The recruitment of innate leukocytes has been well studied in the context of extracellular bacterial and fungal infection but less during viral infections. We have recently shown that the understudied cytokine Interleukin (IL)-17D can mediate neutrophil, natural killer (NK) cell and monocyte infiltration in sterile inflammation and cancer. Herein, we show that early immune cell accumulation at the peritoneal site of infection by mouse cytomegalovirus (MCMV) is mediated by IL-17D. Mice deficient in IL-17D or the transcription factor Nuclear factor (erythroid-derived 2)-like 2 (Nrf2), an inducer of IL-17D, featured an early decreased number of innate immune cells at the point of viral entry and were more susceptible to MCMV infection. Interestingly, we were able to artificially induce innate leukocyte infiltration by applying the Nrf2 activator tert-butylhydroquinone (tBHQ), which rendered mice less susceptible to MCMV infection. Our results implicate the Nrf2/IL-17D axis as a sensor of viral infection and suggest therapeutic benefit in boosting this pathway to promote innate antiviral responses.


Assuntos
Infecções por Herpesviridae/imunologia , Imunidade Inata/imunologia , Interleucina-17/imunologia , Muromegalovirus/imunologia , Fator 2 Relacionado a NF-E2/imunologia , Animais , Linhagem Celular Tumoral , Inibidores Enzimáticos/farmacologia , Hidroquinonas/farmacologia , Interleucina-17/genética , Células Matadoras Naturais/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Muromegalovirus/crescimento & desenvolvimento , Fator 2 Relacionado a NF-E2/genética , Neutrófilos/imunologia , Interferência de RNA , RNA Interferente Pequeno/genética
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