Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 18 de 18
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
2.
FASEB J ; 35(5): e21581, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33871072

RESUMO

Synaptic plasticity damages play a crucial role in the onset and development of depression, especially in the hippocampus, which is more susceptible to stress and the most frequently studied brain region in depression. And, mitochondria have a major function in executing the complex processes of neurotransmission and plasticity. We have previously demonstrated that Iptakalim (Ipt), a new ATP-sensitive potassium (K-ATP) channel opener, could improve the depressive-like behavior in mice. But the underlying mechanisms are not well understood. The present study demonstrated that Ipt reversed depressive-like phenotype in vivo (chronic mild stress-induced mice model of depression) and in vitro (corticosterone-induced cellular model). Further study showed that Ipt could upregulate the synaptic-related proteins postsynaptic density 95 (PSD 95) and synaptophysin (SYN), and alleviated the synaptic structure damage. Moreover, Ipt could reverse the abnormal mitochondrial fission and fusion, as well as the reduced mitochondrial ATP production and collapse of mitochondrial membrane potential in depressive models. Knocking down the mitochondrial ATP-sensitive potassium (Mito-KATP) channel subunit MitoK partly blocked the above effects of Ipt. Therefore, our results reveal that Ipt can alleviate the abnormal mitochondrial dynamics and function depending on MitoK, contributing to improve synaptic plasticity and exert antidepressive effects. These findings provide a candidate compound and a novel target for antidepressive therapy.


Assuntos
Depressão/tratamento farmacológico , Canais KATP/antagonistas & inibidores , Mitocôndrias/efeitos dos fármacos , Propilaminas/farmacologia , Estresse Psicológico/complicações , Sinapses/efeitos dos fármacos , Animais , Depressão/etiologia , Depressão/patologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/metabolismo , Plasticidade Neuronal , Sinapses/metabolismo
3.
Redox Biol ; 41: 101932, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33714739

RESUMO

Pathologically, blood-spinal-cord-barrier (BSCB) disruption after spinal cord injury (SCI) leads to infiltration of numerous peripheral macrophages into injured areas and accumulation around newborn vessels. Among the leaked macrophages, M1-polarized macrophages are dominant and play a crucial role throughout the whole SCI process. The aim of our study was to investigate the effects of M1-polarized bone marrow-derived macrophages (M1-BMDMs) on vascular endothelial cells and their underlying mechanism. Microvascular endothelial cell line bEnd.3 cells were treated with conditioned medium or exosomes derived from M1-BMDMs, followed by evaluations of endothelial-to-mesenchymal transition (EndoMT) and mitochondrial function. After administration, we found conditioned medium or exosomes from M1-BMDMs significantly promoted EndoMT of vascular endothelial cells in vitro and in vivo, which aggravated BSCB disruption after SCI. In addition, significant dysfunction of mitochondria and accumulation of reactive oxygen species (ROS) were also detected. Furthermore, bioinformatics analysis demonstrated that miR-155 is upregulated in both M1-polarized macrophages and microglia. Experimentally, exosomal transfer of miR-155 participated in M1-BMDMs-induced EndoMT and mitochondrial ROS generation in bEnd.3 cells, and subsequently activated the NF-κB signaling pathway by targeting downstream suppressor of cytokine signaling 6 (SOCS6), and suppressing SOCS6-mediated p65 ubiquitination and degradation. Finally, a series of rescue assay further verified that exosomal miR155/SOCS6/p65 axis regulated the EndoMT process and mitochondrial function in vascular endothelial cells. In summary, our work revealed a potential mechanism describing the communications between macrophages and vascular endothelial cells after SCI which could benefit for future research and aid in the development of potential therapies for SCI.


Assuntos
MicroRNAs , Traumatismos da Medula Espinal , Células Endoteliais/metabolismo , Humanos , Recém-Nascido , Macrófagos/metabolismo , MicroRNAs/metabolismo , Mitocôndrias/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais , Traumatismos da Medula Espinal/metabolismo , Proteínas Supressoras da Sinalização de Citocina/metabolismo
4.
Front Plant Sci ; 12: 630644, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33732275

RESUMO

Arabidopsis cyclophilin38 (CYP38) is a thylakoid lumen protein critial for PSII assembly and maintenance, and its C-terminal region serves as the target binding domain. We hypothesized that four conserved residues (R290, F294, Q372, and F374) in the C-terminal domain are critical for the structure and function of CYP38. In yeast two-hybrid and protein pull-down assays, CYP38s with single-sited mutations (R290A, F294A, Q372A, or F374A) did not interact with the CP47 E-loop as the wild-type CYP38. In contrast, CYP38 with the R290A/F294A/Q372A/F374A quadruple mutation could bind the CP47 E-loop. Gene transformation analysis showed that the quadruple mutation prevented CYP38 to efficiently complement the mutant phenotype of cyp38. The C-terminal domain half protein with the quadruple mutation, like the wild-type one, could interact with the N-terminal domain or the CP47 E-loop in vitro. The cyp38 plants expressing CYP38 with the quadruple mutation showed a similar BN-PAGE profile as cyp38, but distinct from the wild type. The CYP38 protein with the quadruple mutation associated with the thylakoid membrane less efficiently than the wild-type CYP38. We concluded that these four conserved residues are indispensable as changes of all these residues together resulted in a subtle conformational change of CYP38 and reduced its intramolecular N-C interaction and the ability to associate with the thylakoid membrane, thus impairing its function in chloroplast.

5.
Eur J Clin Invest ; 51(1): e13443, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33131070

RESUMO

BACKGROUND: To reveal detailed histopathological changes, virus distributions, immunologic properties and multi-omic features caused by SARS-CoV-2 in the explanted lungs from the world's first successful lung transplantation of a COVID-19 patient. MATERIALS AND METHODS: A total of 36 samples were collected from the lungs. Histopathological features and virus distribution were observed by optical microscope and transmission electron microscope (TEM). Immune cells were detected by flow cytometry and immunohistochemistry. Transcriptome and proteome approaches were used to investigate main biological processes involved in COVID-19-associated pulmonary fibrosis. RESULTS: The histopathological changes of the lung tissues were characterized by extensive pulmonary interstitial fibrosis and haemorrhage. Viral particles were observed in the cytoplasm of macrophages. CD3+ CD4- T cells, neutrophils, NK cells, γ/δ T cells and monocytes, but not B cells, were abundant in the lungs. Higher levels of proinflammatory cytokines iNOS, IL-1ß and IL-6 were in the area of mild fibrosis. Multi-omics analyses revealed a total of 126 out of 20,356 significant different transcription and 114 out of 8,493 protein expression in lung samples with mild and severe fibrosis, most of which were related to fibrosis and inflammation. CONCLUSIONS: Our results provide novel insight that the significant neutrophil/ CD3+ CD4- T cell/ macrophage activation leads to cytokine storm and severe fibrosis in the lungs of COVID-19 patient and may contribute to a better understanding of COVID-19 pathogenesis.


Assuntos
COVID-19/patologia , Hemorragia/patologia , Transplante de Pulmão , Pulmão/patologia , Linfonodos/patologia , Fibrose Pulmonar/patologia , Linfócitos B/patologia , Linfócitos B/ultraestrutura , Linfócitos B/virologia , COVID-19/genética , COVID-19/metabolismo , COVID-19/cirurgia , Cromatografia Líquida , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Células Matadoras Naturais/patologia , Células Matadoras Naturais/ultraestrutura , Células Matadoras Naturais/virologia , Pulmão/metabolismo , Pulmão/ultraestrutura , Pulmão/virologia , Linfonodos/metabolismo , Linfonodos/ultraestrutura , Linfonodos/virologia , Macrófagos Alveolares/patologia , Macrófagos Alveolares/ultraestrutura , Macrófagos Alveolares/virologia , Masculino , Pessoa de Meia-Idade , Monócitos/patologia , Monócitos/ultraestrutura , Monócitos/virologia , Neutrófilos/patologia , Neutrófilos/ultraestrutura , Neutrófilos/virologia , Óxido Nítrico Sintase Tipo II/metabolismo , Proteômica , Fibrose Pulmonar/genética , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/cirurgia , RNA-Seq , SARS-CoV-2 , Índice de Gravidade de Doença , Linfócitos T/patologia , Linfócitos T/ultraestrutura , Linfócitos T/virologia , Espectrometria de Massas em Tandem
6.
Mol Ther Nucleic Acids ; 21: 900-915, 2020 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-32810692

RESUMO

Increasing evidence indicates that lymphocyte cytosolic protein 1 (LCP1) overexpression contributes to tumor progression; however, its role in osteosarcoma (OS) remains unclear. We aimed to investigate the potential effect of LCP1 in OS and the underlying mechanisms. We first demonstrated that LCP1 is upregulated in OS cell lines and tissues. Then, we found that aberrant expression of LCP1 could induce the proliferation and metastasis of OS cells in vitro and in vivo by destabilizing neuregulin receptor degradation protein-1 (Nrdp1) and subsequently activating the JAK2/STAT3 signaling pathway. When coculturing OS cells with bone marrow-derived mesenchymal stem cells (BMSCs) in vitro, we validated that oncogenic LCP1 in OS was transferred from BMSCs via exosomes. Moreover, microRNA (miR)-135a-5p, a tumor suppressor, was found to interact upstream of LCP1 to counteract the pro-tumorigenesis effects of LCP1 in OS. In conclusion, BMSC-derived exosomal LCP1 promotes OS proliferation and metastasis via the JAK2/STAT3 pathway. Targeting the miR-135a-5p/LCP1 axis may have potential in treating OS.

7.
J Nanobiotechnology ; 18(1): 105, 2020 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-32711535

RESUMO

BACKGROUND: Spinal cord injury (SCI) is a catastrophic injury that can cause irreversible motor dysfunction with high disability. Exosomes participate in the transport of miRNAs and play an essential role in intercellular communication via transfer of genetic material. However, the miRNAs in exosomes which derived from neurons, and the underlying mechanisms by which they contribute to SCI remain unknown. METHODS: A contusive in vivo SCI model and a series of in vitro experiments were carried out to explore the therapeutic effects of exosomes. Then, a miRNA microarray analysis and rescue experiments were performed to confirm the role of neuron-derived exosomal miRNA in SCI. Western blot, luciferase activity assay, and RNA-ChIP were used to investigate the underlying mechanisms. RESULTS: The results indicated that neuron-derived exosomes promoted functional behavioral recovery by suppressing the activation of M1 microglia and A1 astrocytes in vivo and in vitro. A miRNA array showed miR-124-3p to be the most enriched in neuron-derived exosomes. MYH9 was identified as the target downstream gene of miR-124-3p. A series of experiments were used to confirm the miR-124-3p/MYH9 axis. Finally, it was found that PI3K/AKT/NF-κB signaling cascades may be involved in the modulation of microglia by exosomal miR-124-3p. CONCLUSION: A combination of miRNAs and neuron-derived exosomes may be a promising, minimally invasive approach for the treatment of SCI.


Assuntos
Astrócitos/metabolismo , Exossomos/metabolismo , MicroRNAs , Microglia/metabolismo , Traumatismos da Medula Espinal , Animais , Células Cultivadas , Exossomos/química , Masculino , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/genética , MicroRNAs/metabolismo , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , Neurônios/química , Neurônios/metabolismo , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/fisiopatologia
8.
Acta Biomater ; 103: 196-212, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31857259

RESUMO

Increasing evidence has suggested that paracrine mechanisms might be involved in the underlying mechanism of mesenchymal stem cells (MSCs) transplantation, and exosomes are an important component of this paracrine role. However, MSCs are usually exposed to normoxia (21% O2) in vitro but experience large differences in oxygen concentration in the body under hypoxia. Indeed, hypoxic precondition of MSCs can enhance their paracrine effects. The main purpose of this study was to determine whether exosomes derived from MSCs under hypoxia (Hypo-Exos) exhibit greater effects on bone fracture healing than those under normoxia (Exos). Using in vivo bone fracture model and in vitro experiments including cell proliferation assay, cell migration assay and so on, we confirmed that Hypo-Exos administration promoted angiogenesis, proliferation and migration to a greater extent when compared to Exos. Furthermore, utilizing a series in vitro and in vivo gain and loss of function experiments, we confirmed a functional role for exosomal miR-126 in the process of bone fracture healing. Meanwhile, we found that knockdown of hypoxia inducible factor 1 (HIF-1α) resulted in a significant decrease of miR-126 in MSCs and exosomes, thereby abolishing the effects of Hypo-Exos. In conclusion, our results demonstrated a mechanism by which Hypo-Exos promote bone fracture healing through exosomal miR-126. Moreover, hypoxia preconditioning mediated enhanced production of exosomal miR-126 through the activation of HIF-1α. Hypoxia preconditioning represents an effective and promising method for the optimization of the therapeutic actions of MSC-derived exosomes for bone fracture healing. STATEMENT OF SIGNIFICANCE: Studies have confirmed that transplantation of exosomes exhibit similar therapeutic effects and functional properties to directly-transplanted stem cells but have less significant adverse effects. However, during in vitro culture conditions, MSCs are usually exposed to normoxia (21% O2) which is very different to the oxygen concentrations found in the body under natural physiological conditions. Our results demonstrated a mechanism by which Hypo-Exos promote bone fracture healing through exosomal miR-126 and the SPRED1/Ras/Erk signaling pathway. Moreover, hypoxia preconditioning mediated enhanced production of exosomal miR-126 through the activation of HIF-1α. Hypoxia preconditioning represents an effective and promising method for the optimization of the therapeutic actions of MSC-derived exosomes for bone fracture healing.


Assuntos
Exossomos/metabolismo , Consolidação da Fratura , Fraturas Ósseas/patologia , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Regiões 3' não Traduzidas/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Sequência de Bases , Transplante Ósseo , Hipóxia Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Exossomos/efeitos dos fármacos , Exossomos/ultraestrutura , Consolidação da Fratura/efeitos dos fármacos , Consolidação da Fratura/genética , Fraturas Ósseas/genética , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Camundongos , MicroRNAs/genética , Neovascularização Fisiológica/efeitos dos fármacos , Oxigênio/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos
9.
Biomed Pharmacother ; 116: 109020, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31152928

RESUMO

Glomerulonephritis is the major cause of chronic kidney disease characterized by mesangial cell proliferation and extracellular matrix deposition. The aim of this study was to investigate the effects of Lycium barbarum polysaccharides (LBPs) on anti-Thy 1 nephritis rats and explore the protective mechanism of LBPs. After the model of glomerulonephritis created by injecting anti-thymocyte serum (ATS), rats were treated with enalapril or LBPs for 8 weeks. The therapeutic effect was evaluated by detection of renal-related biochemical parameters, histological observation and markers of renal fibrosis. Moreover, RNA-seq analysis and experiments in vitro were employed to explore the signaling pathway involved in LBPs treatment. The results found that LBPs treatment significantly suppressed ATS-caused increment at levels of blood urea nitrogen, creatinine, proteinuria, PAI-1 protein expression, glomerular mesangial cell proliferation and extracellular matrix hyperplasia, along with reduction of creatinine clearance. RNA sequencing showed pyruvate metabolism acting as a potential signaling pathway, which was evidenced by the inhibitory effect on up-regulation of pyruvate dehydrogenase and PAI-1 levels via treatment with LBPs in vitro. LBPs are the promising agents for the management of glomerulonephritis through pyruvate metabolism signaling pathway.


Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Glomerulonefrite/tratamento farmacológico , Glomerulonefrite/enzimologia , Isoanticorpos/imunologia , Complexo Piruvato Desidrogenase/metabolismo , Animais , Medicamentos de Ervas Chinesas/farmacologia , Fibrose , Glomerulonefrite/genética , Rim/efeitos dos fármacos , Rim/patologia , Rim/fisiopatologia , Rim/ultraestrutura , Testes de Função Renal , Masculino , Camundongos , Piruvatos/metabolismo , Ratos Sprague-Dawley , Coloração e Rotulagem
10.
Adv Healthc Mater ; 7(22): e1800899, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30302950

RESUMO

Antibody-based medicines and nanomedicines are very promising for cancer therapy due to the high specificity and efficacy of antibodies. However, antibody-drug conjugates and antibody-modified nanomaterials frequently suffer from low drug loading and loss of functions due to the covalent modification of the antibody. A novel and versatile strategy to prepare supramolecular nanomaterials by the coassembly of an affibody (antiHER2) and drug-peptide amphiphiles is reported here. During the enzyme-instructed self-assembly process, the drug-peptide amphiphile can coassemble with the affibody, resulting in supramolecular nanofibers in hydrogels. The drug loading in the supramolecular nanofibers is high (>30 wt%), and the stability of antiHER2 is significantly improved in the nanofibers at 37 °C (>15 d in vitro). The supramolecular nanofibers exhibit high affinity for HER2+ cancer cells and can be efficiently taken up by these cells. In a mouse tumor model, the supramolecular nanofibers abolish HER2+ NCI-N87 tumor growth due to the good accumulation and retention of nanofibers in tumor. This study provides a novel strategy to prepare nanomedicines with high drug loading and high specificity.


Assuntos
Antineoplásicos/química , Nanofibras/química , Peptídeos/química , Proteínas Recombinantes de Fusão/química , Sequência de Aminoácidos , Animais , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Hidrogéis/química , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Imagem Óptica , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes de Fusão/uso terapêutico , Transplante Heterólogo
11.
Stem Cells Int ; 2018: 5842714, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30123288

RESUMO

Spermatogonial stem cells (SSCs) are exquisitely regulated to reach a balance between proliferation and differentiation in the niche of seminiferous epithelium. Several extrinsic factors such as GDNF are reported to switch the transition, activating various intrinsic signaling pathways. Transcriptomics analysis could provide a comprehensive landscape of gene expression and regulation. Here, we reanalyzed a previously published transcriptome of two cell types (standing for self-renewing and differentiating SSCs correspondingly). First, we proposed a new parameter, the expression index, to sort the genes considering both absolute and relative expression levels. Using a dynamic statistical model, we identified a list of 1119 candidate genes for SSC self-renewal with the best enrichment of canonical markers. Finally, based on interaction relations, we further optimized the list and constructed a refined network containing integrated information of interactions, expression alternations, biological functions, and disease associations. Further annotation of the 521 refined genes involved in the network revealed an enrichment of well-studied signaling pathways. We believe that the refined network could help us better understand the regulation of SSCs' fates, as well as find novel regulators or targets for SSC self-renewal or preservation of male fertility.

12.
Oncotarget ; 8(31): 51066-51075, 2017 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-28881630

RESUMO

BACKGROUND: Centella asiatica (L.) Urb. is a traditional Chinese medicine that has many medical applications, including wound healing and anti-oxidation. Some traditional Chinese Medicine doctors have found that it has therapeutic effects for external use in the repigmentation of vitiligo and post-inflammatory hypopigmentation. This study was designed to evaluate the effects of madecassoside, a major bioactive component of C. asiatica, on oxidative stress in human melanocytes and its possible mechanism of action. RESULTS: In H2O2-induced oxidative conditions, madecassoside inhibited melanocyte dendrite retraction, improved MMP and reduced the accumulation of [Ca2+]i in a concentration-dependent manner. Observations by TEM showed that madecassoside attenuated the damage of mitochondria in human melanocytes caused by oxidative stress. Furthermore, autophagy activation was demonstrated by AO staining and an increased LC3-II/LC3-I ratio. MATERIALS AND METHODS: Normal human melanocytes were treated with 0.01 mM H2O2 and varying concentrations of madecassoside (0, 10, 50, 100 µg/mL). Subsequently, the retraction velocity of melanocyte dendrites was assessed. Determination of mitochondrial membrane potential (MMP, ΔΨm) was performed by flow cytometry and intracellular calcium ([Ca2+]i) level were measured. Alterations of mitochondrial ultrastructure were observed by transmission electron microscopy (TEM). Acridine orange (AO) staining was used to measure autophagy. The LC3-II/LC3-I ratio, an indicator of autophagosome formation, was analyzed by western blot. CONCLUSIONS: These results demonstrate the antioxidative effect of madecassoside on human melanocytes subjected to oxidative damage via the activation of autophagy. Moreover, madecassoside could be a promising treatment for vitiligo mainly caused by oxidative stress.

13.
Sci Rep ; 7(1): 4603, 2017 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-28676633

RESUMO

Although mesoporous materials and magnetic materials are used to enrich glycopeptides, materials sharing both mesoporous structures and magnetic properties have not been reported for glycopeptide analyses. Here we prepared boronic acid-modified magnetic Fe3O4@mTiO2 microspheres by covalent binding of boronic acid molecules onto the surfaces of silanized Fe3O4@mTiO2 microspheres. The final particles (denoted as B-Fe3O4@mTiO2) showed a typical magnetic hysteresis curve, indicating superparamagnetic behavior; meanwhile, their mesoporous sizes did not change in spite of the reduction in surface area and pore volume. By using these particles together with conventional poly(methyl methacrylate) (PMMA) nanobeads, we then developed a synergistic approach for highly specific and efficient enrichment of N-glycopeptides/glycoproteins. Owing to the introduction of PMMA nanobeads that have strong adsorption towards nonglycopeptides, the number of N-glycopeptides detected and the signal-to-noise ratio in analyzing standard proteins mixture both increased appreciably. The recovery of N-glycopeptides by the synergistic method reached 92.1%, much improved than from B-Fe3O4@mTiO2 alone that was 75.3%. Finally, we tested this approach in the analysis of amniotic fluid, obtaining the maximum number and ratio of N-glycopeptides compared to the use of B-Fe3O4@mTiO2 alone and commercial SiMAG-boronic acid particles. This ensemble provides an interesting and efficient enrichment platform for glycoproteomics research.


Assuntos
Líquido Amniótico/química , Ácidos Borônicos/química , Glicopeptídeos/análise , Titânio/química , Adsorção , Adulto , Cromatografia Líquida , Feminino , Humanos , Idade Materna , Microesferas , Polimetil Metacrilato/química , Espectrometria de Massas em Tandem
14.
Arch Dermatol Res ; 309(5): 323-333, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28314912

RESUMO

Skin color is determined by the number of melanin granules produced by melanocytes that are transferred to keratinocytes. Melanin synthesis and the distribution of melanosomes to keratinocytes within the epidermal melanin unit (EMU) within the skin of vitiligo patients have been poorly studied. The ultrastructure and distribution of melanosomes in melanocytes and surrounding keratinocytes in perilesional vitiligo and normal skin were investigated using transmission electron microscopy (TEM). Furthermore, we performed a quantitative analysis of melanosome distribution within the EMUs with scatter plot. Melanosome count within keratinocytes increased significantly compared with melanocytes in perilesional stable vitiligo (P < 0.001), perilesional halo nevi (P < 0.01) and the controls (P < 0.01), but not in perilesional active vitiligo. Furthermore, melanosome counts within melanocytes and their surrounding keratinocytes in perilesional active vitiligo skin decreased significantly compared with the other groups. In addition, taking the means-standard error of melanosome count within melanocytes and keratinocytes in healthy controls as a normal lower limit, EMUs were graded into 3 stages (I-III). Perilesional active vitiligo presented a significantly different constitution in stages compared to other groups (P < 0.001). The distribution and constitution of melanosomes were normal in halo nevi. Impaired melanin synthesis and melanosome transfer are involved in the pathogenesis of vitiligo. Active vitiligo varies in stages and in stage II, EMUs are slightly impaired, but can be resuscitated, providing a golden opportunity with the potential to achieve desired repigmentation with an appropriate therapeutic choice. Adverse milieu may also contribute to the low melanosome count in keratinocytes.


Assuntos
Queratinócitos/metabolismo , Melanossomas/metabolismo , Nevo com Halo/patologia , Pigmentação da Pele/fisiologia , Vitiligo/patologia , Adolescente , Adulto , Células Epidérmicas , Epiderme/patologia , Feminino , Humanos , Masculino , Melaninas/metabolismo , Melanócitos/patologia , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Adulto Jovem
15.
PLoS One ; 11(3): e0152067, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27003708

RESUMO

MaxQuant is a proteomic software widely used for large-scale tandem mass spectrometry data. We have designed and developed an enhanced result reporting tool for MaxQuant, named as MaxReport. This tool can optimize the results of MaxQuant and provide additional functions for result interpretation. MaxReport can generate report tables for protein N-terminal modifications. It also supports isobaric labelling based relative quantification at the protein, peptide or site level. To obtain an overview of the results, MaxReport performs general descriptive statistical analyses for both identification and quantification results. The output results of MaxReport are well organized and therefore helpful for proteomic users to better understand and share their data. The script of MaxReport, which is freely available at http://websdoor.net/bioinfo/maxreport/, is developed using Python code and is compatible across multiple systems including Windows and Linux.


Assuntos
Proteoma/metabolismo , Proteômica/métodos , Bases de Dados de Proteínas , Peptídeos/metabolismo , Software , Espectrometria de Massas em Tandem/métodos
16.
Arch Dermatol Res ; 307(3): 281-9, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25672813

RESUMO

Vitiligo and halo nevi are both pigmentary disorders of the skin characterized by the acquired loss of functional epidermal melanocytes manifesting as white macules and patches. The cellular mechanism(s) and biochemical changes that result in the appearance of these two types of achromic lesions are still uncertain; and the relationship between vitiligo and halo nevi has been in dispute. In this study, we investigated the ultrastructure of mitochondria in melanocytes and in keratinocytes from perilesional vitiligo skin and from perilesional halo nevi skin using Transmission Electron Microscopy. Furthermore, we performed a quantitative analysis of mitochondrial morphology through a stereological study. As previously reported, we found that melanocytes from perilesional active vitiligo skin were loosely connected with their surroundings by their retracted dendrites. The surface density and the volume density of mitochondria in melanocytes and in keratinocytes from perilesional vitiligo skin are increased significantly compared with the controls, especially in active vitiligo. In contrast, there are no significant differences in mitochondria in melanocytes and in keratinocytes from perilesional halo nevi skin compared with the controls. In summary, the tendency of different morphologic alterations in mitochondria from perilesional vitiligo skin and from perilesional halo nevi skin reflect heterogeneous backgrounds between the two diseases, revealing that vitiligo and halo nevi may have separate pathogenic mechanisms. These findings may help elucidate the relationship of these two diseases and their underlying mechanisms.


Assuntos
Queratinócitos/ultraestrutura , Melanócitos/ultraestrutura , Mitocôndrias/ultraestrutura , Nevo com Halo/diagnóstico , Vitiligo/diagnóstico , Adolescente , Adulto , Células Cultivadas , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Pele/patologia , Adulto Jovem
17.
J Dermatol Sci ; 77(2): 117-24, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25592908

RESUMO

BACKGROUND: Vitiligo is an acquired pigmentary disorder of unknown etiology that is clinically characterized by the development of white macules in the skin related to the selective loss of melanocytes in those areas. Evidence shows that mitochondria might be a unifying target of reactive oxygen species (ROS) generation, cytokine production, catecholamine release and/or alteration of Ca(2+) metabolism that leads to melanocyte loss. OBJECTIVE: To assess the protective effect of calcipotriol on mitochondria of human melanocytes by investigating their dendritic morphology under oxidative stress. METHODS: Human melanocytes were treated with 0.05% H2O2 as well as various concentrations of calcipotriol, after which the retraction velocity of melanocyte dendrites was assessed. Detection of malondialdehyde (MDA) and superoxide dismutase (SOD) was performed as were the mitochondrial membrane potential (MMP) and intracellular calcium concentration ([Ca(2+)]i). Ultrastructural changes of mitochondria in melanocytes were observed by transmission electron microscopy. In addition, the expression of Beclin1, microtubule-associated protein 1 light chain 3 (LC3), dynamin related protein 1 (Drp1), mitofusin 1 (Mfn1) and mitofusin 2 (Mfn2), which are related to autophagy and mitochondrial dynamics, were analyzed by Western blot. RESULTS: Calcipotriol reduced the retraction velocity of melanocyte dendrites. In addition, calcipotriol, from 20nM to 80nM, decreased the level of MDA, increased the activity of SOD, suppressed the reduction of MMP and recovered Ca(2+) homeostasis by reducing [Ca(2+)]i in a concentration-dependent manner. Observation by transmission electron microscopy suggested that calcipotriol might reduce the injury of mitochondria in melanocytes under oxidative stress. Furthermore, the expression of Beclin1, LC3-II/LC3-I, Mfn2 and Drp1 was higher in the calcipotriol-treated melanocytes than in the control or H2O2-treated melanocytes. The level of Mfn1 was almost unchanged, but was higher at a concentration of 80nM calcipotriol than in any other condition. The expression of Mfn2 and Drp1 decreased with increasing calcipotriol concentration. CONCLUSION: Our study demonstrates the antioxidative effect of calcipotriol on melanocytes against oxidative damage. Moreover, calcipotriol could be a promising drug delivery strategy to protect melanocytes against oxidative damage in vitiligo through autophagy or mitophagy.


Assuntos
Calcitriol/análogos & derivados , Fármacos Dermatológicos/farmacologia , Melanócitos/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Western Blotting/métodos , Calcitriol/farmacologia , Cálcio/análise , Criança , Circuncisão Masculina , Células Dendríticas , Citometria de Fluxo/métodos , Prepúcio do Pênis/ultraestrutura , Humanos , Masculino , Melanócitos/ultraestrutura , Microscopia Eletrônica de Transmissão/métodos , Mitocôndrias/ultraestrutura , Mitofagia/efeitos dos fármacos , Espécies Reativas de Oxigênio/efeitos adversos , Vitiligo/etiologia , Vitiligo/patologia
18.
Biol Reprod ; 85(3): 635-42, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21565992

RESUMO

This study compared the ultrastructural differences of term placentae from human pregnancies resulting from assisted reproductive technology (ART) with term placentae from spontaneous human pregnancies. Term placentae were taken from women who had undergone an ART procedure (n = 8) and matched with term placentae from women who had had a spontaneous pregnancy (controls, n = 15). Using light microscopy (LM) and transmission-electron microscopy (TEM), terminal villi were evaluated with respect to the placental blood barrier, fetal capillaries, villous stroma, as well as cytotrophoblasts and syncytiotrophoblasts (ST) along with their substructures. No obvious differences were found between the ART-derived and control placentae when LM was used. With TEM, however, differences in the ultrastructural features were seen in the ART-derived placentae, specifically degenerative alterations of the terminal villi, mainly in ST, including a thicker placental barrier, decreased apical microvilli, and increased multiple vacuoles. The results demonstrate that some ultrastructural differences exist between ART-derived and control placentae with respect to the placental blood barrier, which may suggest maternofetal traffic downregulation following ART treatment. Further studies are required to understand the ultrastructural changes and their potential functional aspects in ART pregnancies.


Assuntos
Placenta/ultraestrutura , Técnicas de Reprodução Assistida/efeitos adversos , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Microscopia Eletrônica de Transmissão , Placenta/irrigação sanguínea , Gravidez , Adulto Jovem
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...