Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 4.509
Filtrar
1.
Int J Nanomedicine ; 16: 3707-3724, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34103912

RESUMO

Introduction: Intracellular delivery of molecules is central to applications in biotechnology, medicine, and basic research. Nanoparticle-mediated photoporation using carbon black nanoparticles exposed to pulsed, near-infrared laser irradiation offers a physical route to create transient cell membrane pores, enabling intracellular delivery. However, nanoparticle-mediated photoporation, like other physical intracellular delivery technologies, necessitates a trade-off between achieving efficient uptake of exogenous molecules and maintaining high cell viability. Methods: In this study, we sought to shift this balance by adding serum to cells during nanoparticle-mediated photoporation as a viability protectant. DU-145 prostate cancer cells and human dermal fibroblasts were exposed to laser irradiation in the presence of carbon black (CB) nanoparticles and other formulation additives, including fetal bovine serum (FBS) and polymers. Results: Our studies showed that FBS can protect cells from viability loss, even at high-fluence laser irradiation conditions that lead to high levels of intracellular delivery in two different mammalian cell types. Further studies revealed that full FBS was not needed: viability protection was achieved with denatured FBS, with just the high molecular weight fraction of FBS (>30 kDa), or even with individual proteins like albumin or hemoglobin. Finally, we found that viability protection was also obtained using certain neutral water-soluble polymers, including Pluronic F127, polyvinylpyrrolidone, poly(2-ethyl-2-oxazoline), and polyethylene glycol, which were more effective at increased concentration, molecular weight, or hydrophobicity. Conclusion: Altogether, these findings suggest an interaction between amphiphilic domains of polymers with the cell membrane to help cells maintain viability, possibly by facilitating transmembrane pore closure. In this way, serum components or synthetic polymers can be used to increase intracellular delivery by nanoparticle-mediated photoporation while maintaining high cell viability.


Assuntos
Citoproteção , Sistemas de Liberação de Medicamentos , Espaço Intracelular/química , Luz , Nanopartículas/química , Soro/química , Carboximetilcelulose Sódica/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos da radiação , Citoproteção/efeitos da radiação , Fibroblastos/efeitos da radiação , Humanos , Lasers , Peso Molecular , Poloxâmero/química , Polietilenoglicóis/química , Fuligem/química , Viscosidade
2.
Nat Commun ; 12(1): 2742, 2021 05 12.
Artigo em Inglês | MEDLINE | ID: mdl-33980846

RESUMO

Ultraviolet radiation (UVR) damages the dermis and fibroblasts; and increases melanoma incidence. Fibroblasts and their matrix contribute to cancer, so we studied how UVR modifies dermal fibroblast function, the extracellular matrix (ECM) and melanoma invasion. We confirmed UVR-damaged fibroblasts persistently upregulate collagen-cleaving matrix metalloprotein-1 (MMP1) expression, reducing local collagen (COL1A1), and COL1A1 degradation by MMP1 decreased melanoma invasion. Conversely, inhibiting ECM degradation and MMP1 expression restored melanoma invasion. Primary cutaneous melanomas of aged humans show more cancer cells invade as single cells at the invasive front of melanomas expressing and depositing more collagen, and collagen and single melanoma cell invasion are robust predictors of poor melanoma-specific survival. Thus, primary melanomas arising over collagen-degraded skin are less invasive, and reduced invasion improves survival. However, melanoma-associated fibroblasts can restore invasion by increasing collagen synthesis. Finally, high COL1A1 gene expression is a biomarker of poor outcome across a range of primary cancers.


Assuntos
Colágeno/metabolismo , Melanoma/metabolismo , Melanoma/terapia , Raios Ultravioleta , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Ensaio de Imunoadsorção Enzimática , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Lentivirus/genética , Espectrometria de Massas , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 1 da Matriz/metabolismo , Microscopia de Força Atômica
3.
Int J Mol Sci ; 22(8)2021 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-33917737

RESUMO

Cellular senescence and aging result in a reduced ability to manage persistent types of inflammation. Thus, the chronic low-level inflammation associated with aging phenotype is called "inflammaging". Inflammaging is not only related with age-associated chronic systemic diseases such as cardiovascular disease and diabetes, but also skin aging. As the largest organ of the body, skin is continuously exposed to external stressors such as UV radiation, air particulate matter, and human microbiome. In this review article, we present mechanisms for accumulation of senescence cells in different compartments of the skin based on cell types, and their association with skin resident immune cells to describe changes in cutaneous immunity during the aging process.


Assuntos
Envelhecimento/fisiologia , Microambiente Celular , Senescência Celular , Inflamação/etiologia , Pele/metabolismo , Animais , Senescência Celular/genética , Senescência Celular/efeitos da radiação , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Inflamação/patologia , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Melanócitos/metabolismo , Melanócitos/efeitos da radiação , Pele/citologia , Fenômenos Fisiológicos da Pele , Pigmentação da Pele
4.
Ann Agric Environ Med ; 28(1): 163-171, 2021 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-33775083

RESUMO

INTRODUCTION: Currently, mobile phones and Wi-Fi are the most commonly used forms of telecommunication. The popularity of mobile telecommunications has made it necessary to investigate the problem more comprehensively and cautiously assess the possible risks, because never before in history has such a substantial proportion of the population been exposed to microwaves at comparably high levels. Some studies indicate that the high frequency electromagnetic radiation emitted by mobile phone and Wi-Fi connections can have a negative effect on human health, and can cause cancer. OBJECTIVE: The aim of the study was to investigate the influence of the radiofrquency electromagnetic field (RF-EMF) on the metaboloc activity and morphology of normal human cells (fibroblasts) and cancer cells (prostate cancer cells). MATERIAL AND METHODS: The cell cultures (human fibroblasts and prostate cancer cells) were exposed to RF-EMF at the frequency of 2.5 GHz for 24, 48 and 72h. To quantify changes in cell viability, the Cell Counting Kit - 8 was used. RESULTS: It was found that the RF electromagnetic field exposure caused a significant decrease in the viability of fibroblasts, and a significant increase in cancer cells. Morphological analysis did not show significant changes in both cell lines after exposure to RF-EMF. CONCLUSIONS: On the basis of the obtained results, the hypothesis can be formulated that a high frequency electromagnetic field can have harmful effects on human cells.


Assuntos
Linhagem Celular Tumoral/efeitos da radiação , Campos Eletromagnéticos/efeitos adversos , Fibroblastos/efeitos da radiação , Ondas de Rádio/efeitos adversos , Linhagem Celular , Telefone Celular , Sobrevivência Celular/efeitos da radiação , Exposição Ambiental/efeitos adversos , Fibroblastos/citologia , Humanos
5.
Int J Mol Sci ; 22(4)2021 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-33669634

RESUMO

Little is known about the effects on hyaluronan (HA) metabolism of UVA radiation. This study demonstrates that the secretion of HA by human dermal fibroblasts (HDFs) is downregulated by UVA, accompanied by the down- and upregulation of mRNA and protein levels of the HA-synthesizing enzyme (HAS2) and the HA-degrading protein, HYaluronan Binding protein Involved in HA Depolymerization(HYBID), respectively. Signaling analysis revealed that the exposure distinctly elicits activation of the p38/MSK1/CREB/c-Fos/AP-1 axis, the JNK/c-Jun axis, and the p38/ATF-2 axis, but downregulates the phosphorylation of NF-kB and JAK/STAT3. A signal inhibition study demonstrated that the inhibition of p38 significantly abrogates the UVA-accentuated mRNA level of HYBID. Furthermore, the inhibition of STAT3 significantly downregulates the level of HAS2 mRNA in non-UVA exposed HDFs. Analysis using siRNAs demonstrated that transfection of ATF-2 siRNA but not c-Fos siRNA abrogates the increased protein level of HYBID in UVA-exposed HDFs. An inhibitor of protein tyrosine phosphatase but not of protein serine/threonine phosphatase restored the diminished phosphorylation level of STAT3 at Tyr 705, accompanied by a significant abolishing effect on the decreased mRNA expression level of HAS2. Silencing with a protein tyrosine phosphatase PTP-Meg2 siRNA revealed that it abrogates the decreased phosphorylation of STAT3 at Tyr 705 in UVA-exposed HDFs. These findings suggest that the UVA-induced decrease in HA secretion by HDFs is attributable to the down- and upregulation of HAS2 and HYBID expression, respectively, changes that are mainly ascribed to the inactivated signaling of the STAT3 axis due to the activated tyrosine protein phosphatase PTP-Meg2 and the activated signaling of the p38/ATF2 axis, respectively.


Assuntos
Regulação para Baixo/efeitos da radiação , Fibroblastos/efeitos da radiação , Hialuronan Sintases/metabolismo , Ácido Hialurônico/metabolismo , Hialuronoglucosaminidase/metabolismo , Transdução de Sinais/efeitos da radiação , Raios Ultravioleta , Regulação para Cima/efeitos da radiação , Fator 2 Ativador da Transcrição/metabolismo , Morte Celular/efeitos dos fármacos , Morte Celular/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Derme/citologia , Regulação para Baixo/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Janus Quinase 2/metabolismo , Masculino , Modelos Biológicos , Peso Molecular , Fosforilação/efeitos dos fármacos , Fosforilação/efeitos da radiação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Transcrição STAT3/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
6.
PLoS Genet ; 17(1): e1009302, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33444353

RESUMO

Human skin is continuously exposed to environmental DNA damage leading to the accumulation of somatic mutations over the lifetime of an individual. Mutagenesis in human skin cells can be also caused by endogenous DNA damage and by DNA replication errors. The contributions of these processes to the somatic mutation load in the skin of healthy humans has so far not been accurately assessed because the low numbers of mutations from current sequencing methodologies preclude the distinction between sequencing errors and true somatic genome changes. In this work, we sequenced genomes of single cell-derived clonal lineages obtained from primary skin cells of a large cohort of healthy individuals across a wide range of ages. We report here the range of mutation load and a comprehensive view of the various somatic genome changes that accumulate in skin cells. We demonstrate that UV-induced base substitutions, insertions and deletions are prominent even in sun-shielded skin. In addition, we detect accumulation of mutations due to spontaneous deamination of methylated cytosines as well as insertions and deletions characteristic of DNA replication errors in these cells. The endogenously induced somatic mutations and indels also demonstrate a linear increase with age, while UV-induced mutation load is age-independent. Finally, we show that DNA replication stalling at common fragile sites are potent sources of gross chromosomal rearrangements in human cells. Thus, somatic mutations in skin of healthy individuals reflect the interplay of environmental and endogenous factors in facilitating genome instability and carcinogenesis.


Assuntos
Dano ao DNA/efeitos da radiação , Metilação de DNA/genética , Replicação do DNA/genética , Pele/efeitos da radiação , Metilação de DNA/efeitos da radiação , Reparo do DNA/efeitos da radiação , Replicação do DNA/efeitos da radiação , Fibroblastos/efeitos da radiação , Genoma Humano/genética , Genoma Humano/efeitos da radiação , Instabilidade Genômica/efeitos da radiação , Genômica/métodos , Humanos , Mutação INDEL/efeitos da radiação , Melanócitos/efeitos da radiação , Mutagênese/genética , Mutagênese/efeitos da radiação , Pele/metabolismo , Raios Ultravioleta/efeitos adversos
7.
Methods Mol Biol ; 2239: 213-234, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33226622

RESUMO

Mouse embryonic fibroblasts (MEFs) can be used in co-culture to support generation of induced pluripotent stem cells (iPSCs) and the normal growth and proliferation of human pluripotent stem cells (hPSCs). Here, we describe the necessary steps to derive, expand, harvest, inactivate, plate, and use MEFs as feeders for iPSC generation and maintenance.


Assuntos
Técnicas de Cultura de Células/métodos , Embrião de Mamíferos/citologia , Células Alimentadoras/efeitos da radiação , Fibroblastos/efeitos da radiação , Animais , Células Cultivadas , Técnicas de Cocultura/métodos , Criopreservação , Células Alimentadoras/citologia , Células Alimentadoras/fisiologia , Fibroblastos/citologia , Fibroblastos/fisiologia , Células-Tronco Pluripotentes Induzidas/fisiologia , Camundongos
8.
J Photochem Photobiol B ; 214: 112100, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33316625

RESUMO

Photobiomodulation (PBM) is reported to impart a range of clinical benefits, from the healing of chronic wounds to athletic performance enhancement. The increasing prevalence of this therapy conflicts with the lack of understanding concerning specific cellular mechanisms induced by PBM. Herein, we systematically explore the literature base, specifically related to PBM (within the range 600-1070 nm) and its influence on dermal fibroblasts. The existing research in this field is appraised through five areas: cellular proliferation and viability; cellular migration; ATP production and mitochondrial membrane potential; cellular protein expression and synthesis; and gene expression. This review demonstrates that when fibroblasts are irradiated in vitro within a set range of intensities, they exhibit a multitude of positive effects related to the wound healing process. However, the development of an optimal in vitro framework is paramount to improve the reliability and validity of research in this field.


Assuntos
Fibroblastos/efeitos da radiação , Terapia com Luz de Baixa Intensidade/métodos , Cicatrização/efeitos da radiação , Linhagem Celular , Movimento Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Sobrevivência Celular/efeitos da radiação , Fibroblastos/citologia , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Técnicas In Vitro , Proteínas/genética , Proteínas/metabolismo , Pele
9.
J Oleo Sci ; 69(11): 1487-1495, 2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-33055443

RESUMO

Photoaged skin is characterized by the appearance of pigmented spots such as solar lentigos, deep wrinkles and sags, and progresses due to chronic sun exposure. Among the wavelengths of sunlight, UVA is responsible for the appearance of wrinkles and sags that originate from structural alterations in the dermis of photoaged skin such as the depletion of collagen fibers. Thus, improving and restoring collagen fibers is an effective approach to reduce skin photoaging and maintain a youthful appearance. This study was conducted to evaluate the potential of an extract of Ocimum basilicum (OC), which contains rosmarinic acid (RA), as an anti-photoaging material focusing on the capacity to restore collagen fibers that are disrupted due to intracellular oxidative stress. In spite of their relatively low capacities for chemical scavenging of reactive oxygen species (ROS), both OC and RA showed efficient removal of biological oxidative stress by reducing levels of intracellular ROS and carbonylated proteins (CPs) in fibroblasts following exposure to single or repetitive UVA irradiations. Fibroblasts irradiated with repetitive UVA as a model for chronic sun-exposed cells showed significant increases in matrix metalloproteinase-1 and decreases in type I collagen synthesis and formed reduced numbers of collagen fibers. Since both OC and RA restored the adverse phenomena caused by repetitive UVA irradiation, we conclude that OC containing RA is an effective anti-photoaging material.


Assuntos
Cinamatos/farmacologia , Colágeno/metabolismo , Colágeno/efeitos da radiação , Depsídeos/farmacologia , Derme/citologia , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Ocimum basilicum/química , Extratos Vegetais/farmacologia , Envelhecimento da Pele/efeitos dos fármacos , Raios Ultravioleta/efeitos adversos , Células Cultivadas , Cinamatos/isolamento & purificação , Depsídeos/isolamento & purificação , Fibroblastos/patologia , Humanos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Espécies Reativas de Oxigênio/metabolismo , Envelhecimento da Pele/patologia
10.
Arch Biochem Biophys ; 689: 108437, 2020 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-32526201

RESUMO

Skin aging is influenced by several genetic, physiological, and environmental factors. In particular, ultraviolet (UV) exposure is an important factor involved in inducing skin photoaging. Autophagy controlling homeostatic balance between the synthesis, degradation, and recycling of cellular organelles and proteins plays important regulatory roles in several biological processes, including aging. The opioid neuropeptide α-neoendorphin (named NEP) is an endogenous decapeptide (N-YGGFLRKYPK-C) that activates the kappa opioid receptor and exhibits certain anti-aging and anti-wrinkling effects on skin cells; however, its action mechanism has not yet been elucidated. Therefore, the aim of this study was to determine the effects of NEP on anti-skin aging and autophagy activation in human dermal fibroblast cells. Western blot results showed that NEP down-regulates the production of phospho-mammalian target of rapamycin (p-mTOR), whereas increases the expression of key autophagy-related molecules such as Beclin-1, Atg5-Atg12, and LC3-II. The immunocytochemical analysis performed with anti-LC3-II antibody also showed that the autophagic indicators, autophagosomes are formed by NEP. These results suggest that NEP can activate cellular autophagy through mTOR-Beclin-1-mediated signaling pathway. It was also revealed by CM-H2DCF-DA assay and Western blottings that NEP can reduce the production of ultraviolet B (UVB)-induced reactive oxygen species (ROS) like with N-acetylcysteine (NAC), resulting in decreasing the expression levels of skin aging-related proteins, such as phospho-ERK (p-ERK), phospho-p38 (p-p38), and phospho-JNK (p-JNK). Furthermore, NEP could increase the type I procollagen production, while decreasing MMP-1, MMP-2, and MMP-9 activities. Taken together, the results demonstrate that NEP can reduce UVB-induced photoaging by activating autophagy.


Assuntos
Autofagia , Endorfinas/metabolismo , Precursores de Proteínas/metabolismo , Envelhecimento da Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Linhagem Celular , Derme/citologia , Derme/metabolismo , Derme/efeitos da radiação , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Pró-Colágeno/metabolismo , Espécies Reativas de Oxigênio/metabolismo
11.
Chin J Nat Med ; 18(6): 436-445, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32503735

RESUMO

This study investigated the effects of X-ray irradiation on primary rat cardiac fibroblasts (CFs) and its potential mechanism, as well as whether sodium tanshinone IIA sulfonate (STS) has protective effect on CFs and its possible mechanism. Our data demonstrated that X-rays inhibited cell growth and increased oxidative stress in CFs, and STS mitigated X-ray-induced injury. Enzyme-linked immuno-sorbent assay showed that X-rays increased the levels of secreted angiotensin II (Ang II) and brain natriuretic peptide (BNP). STS inhibited the X-ray-induced increases in Ang II and BNP release. Apoptosis and cell cycle of CFs were analyzed using flow cytometry. X-rays induced apoptosis in CFs, whereas STS inhibited apoptosis in CFs after X-ray irradiation. X-rays induced S-phase cell cycle arrest in CFs, which could be reversed by STS. X-rays increased the expression of phosphorylated-P38/P38, cleaved caspase-3 and caspase-3 as well as decreased the expression of phosphorylated extracellular signal-regulated kinase 1/2 (ERK 1/2)/ERK 1/2 and B cell lymphoma 2 (Bcl-2)/Bcl-2 associated X protein (BAX) in CFs, as shown by Western blotting. STS mitigated the X-ray radiation-induced expression changes of these proteins. In conclusion, our results demonstrated that STS may potentially be developed as a medical countermeasure to mitigate radiation-induced cardiac damage.


Assuntos
Fibroblastos/efeitos dos fármacos , Fibroblastos/efeitos da radiação , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/efeitos da radiação , Fenantrenos/farmacologia , Lesões por Radiação/prevenção & controle , Animais , Apoptose/efeitos dos fármacos , Células Cultivadas , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
12.
Br J Radiol ; 93(1112): 20190949, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32464080

RESUMO

OBJECTIVES: The relative biological effectiveness (RBE) of X-rays and γ radiation increases substantially with decreasing beam energy. This trend affects the efficacy of medical applications of this type of radiation. This study was designed to develop a model based on a survey of experimental data that can reliably predict this trend. METHODS: In our model, parameters α and ß of a cell survival curve are simple functions of the frequency-average linear energy transfer (LF) of delta electrons. The choice of these functions was guided by a microdosimetry-based model. We calculated LF by using an innovative algorithm in which LF is associated with only those electrons that reach a sensitive-to-radiation volume (SV) within the cell. We determined model parameters by fitting the model to 139 measured (α,ß) pairs. RESULTS: We tested nine versions of the model. The best agreement was achieved with [Formula: see text] and ß being linear functions of [Formula: see text] .The estimated SV diameter was 0.1-1 µm. We also found that α, ß, and the α/ß ratio increased with increasing [Formula: see text] . CONCLUSIONS: By combining an innovative method for calculating [Formula: see text] with a microdosimetric model, we developed a model that is consistent with extensive experimental data involving photon energies from 0.27 keV to 1.25 MeV. ADVANCES IN KNOWLEDGE: We have developed a photon RBE model applicable to an energy range from ultra-soft X-rays to megaelectron volt γ radiation, including high-dose levels where the RBE cannot be calculated as the ratio of α values. In this model, the ionization density represented by [Formula: see text] determines the RBE for a given photon spectrum.


Assuntos
Sobrevivência Celular/efeitos da radiação , Raios gama , Modelos Estatísticos , Raios X , Animais , Linhagem Celular , Cricetulus , Relação Dose-Resposta à Radiação , Fibroblastos/efeitos da radiação , Humanos , Camundongos , Doses de Radiação
13.
J Photochem Photobiol B ; 205: 111824, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32126496

RESUMO

Excessive exposure to UVB radiation can lead to oxidative and inflammatory damage that compromises the cutaneous integrity. The application on the skin of photochemoprotective products is considered a relevant approach for the prevention of oxidative damage. In this study the in vitro and in vivo photochemoprotective effects of antioxidant plant materials obtained from the leaves of Nectandra cuspidata Nees following UVB irradiation were evaluated. The cytoprotective effect, reactive oxygen species (ROS) production and lipid peroxidation (LPO) were assessed in L-929 fibroblasts treated with the ethyl acetate fraction (EAF) or isolated compounds (epicatechin, isovitexin and vitexin) before or after irradiation with UVB (500 mJ/cm2). EAF substantially reduced the dead of cells and inhibited the UVB-induced ROS production and LPO in both treatments, compared with the irradiated untreated fibroblasts, presenting effects similar or better than pure compounds. The in vivo photochemoprotective effects of a topical emulsion containing 1% EAF (F2) were evaluated in hairless mice exposed to UVB. F2 improved all evaluated parameters in the skin of animals, inhibited ROS production, increased antioxidant defenses by decreasing reduced glutathione (GSH) and catalase depletion, reduced the activities of metalloproteinases (MMP-2 and MMP-9) and myeloperoxidase, decreased epidermal thickness and skin edema, and inhibited the appearance of sunburn cells as well as the recruitment of neutrophils and mast cell inflammatory infiltrates. These findings show that EAF presents high photochemoprotective effects, and that a topical formulation containing it may have potential for skin care.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Fibroblastos/efeitos dos fármacos , Lauraceae , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Protetores contra Radiação/farmacologia , Pele/efeitos dos fármacos , Raios Ultravioleta/efeitos adversos , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Feminino , Fibroblastos/efeitos da radiação , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Camundongos Pelados , Folhas de Planta , Espécies Reativas de Oxigênio/metabolismo , Pele/metabolismo , Pele/patologia , Pele/efeitos da radiação
14.
Radiat Res ; 193(5): 451-459, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32150497

RESUMO

Findings from previous studies have suggested that the telomerase system is involved in radiation-induced genomic instability. In this study, we investigated the involvement of telomerase in the development and processing of chromosomal damage at different cell cycle stages after irradiation of human fibroblasts. Several response criteria were investigated, including cell survival, chromosomal damage (using the micronucleus assay), G2-induced chromatid aberrations (using the conventional G2 assay as well as a chemically-induced premature chromosome condensation assay) and DNA double-strand breaks (DSBs; using γ-H2AX, 53BP1 and Rad51) in an isogenic pair of cell lines: BJ human foreskin fibroblasts and BJ1-hTERT, a telomerase-immortalized BJ cell line. To distinguish among G1, S and G2 phase, cells were co-immunostained for CENP-F and cyclin A, which are tightly regulated proteins in the cell cycle. After X-ray irradiation at doses in the range of 0.1-6 Gy, the results showed that for cell survival and micronuclei induction, where the overall effect is dominated by the cells in G1 and S phase, no difference was observed between the two cell types; in contrast, when radiation sensitivity at the G2 stage of the cell cycle was analyzed, a significantly higher number of chromatid-type aberrations (breaks and exchanges), and higher levels of γ-H2AX and of Rad51 foci were observed for the BJ cells compared to the BJ1-hTERT cells. Therefore, it can be concluded that telomerase appears to be involved in DNA DSB repair processes, mainly in the G2 phase. These data, taken overall, reinforce the notion that hTERT or other elements of the telomere/telomerase system may defend chromosome integrity in human fibroblasts by promoting repair in G2 phase of the cell cycle.


Assuntos
Instabilidade Genômica/efeitos da radiação , Telomerase/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos da radiação , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Fase G2/efeitos da radiação , Raios gama/efeitos adversos , Humanos , Testes para Micronúcleos , Rad51 Recombinase/metabolismo , Fase S/efeitos da radiação
15.
Int J Mol Sci ; 21(3)2020 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-32024276

RESUMO

Skin is being increasingly exposed to artificial blue light due to the extensive use of electronic devices. This, together with recent observations reporting that blue light-also known as high-energy visible light-can exert cytotoxic effects associated with oxidative stress and promote hyperpigmentation, has sparked interest in blue light and its potential harmful effects on skin. The photoprotective properties of new extracts of different botanicals with antioxidant activity are therefore being studied. Deschampsia antarctica (Edafence®, EDA), a natural aqueous extract, has shown keratinocyte and fibroblast cell protection effects against ultraviolet radiation and dioxin toxicity. In this regard, we studied the protective capacity of EDA against the deleterious effects of artificial blue light irradiation in human dermal fibroblasts (HDF) and melanocytes. We analyzed the impact of EDA on viability, cell morphology, oxidative stress, melanogenic signaling pathway activation and hyperpigmentation in HDF and melanocytes subjected to artificial blue light irradiation. Our results show that EDA protects against cell damage caused by artificial blue light, decreasing oxidative stress, melanogenic signaling pathway activation and hyperpigmentation caused by blue light irradiation. All these findings suggest that EDA might help prevent skin damage produced by artificial blue light exposure from screen of electronic devices.


Assuntos
Luz/efeitos adversos , Extratos Vegetais/farmacologia , Poaceae/química , Substâncias Protetoras/farmacologia , Envelhecimento da Pele/efeitos dos fármacos , Pele/efeitos dos fármacos , Dispositivos Eletrônicos Vestíveis/efeitos adversos , Sobrevivência Celular , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/efeitos da radiação , Humanos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Queratinócitos/efeitos da radiação , Estresse Oxidativo , Espécies Reativas de Oxigênio , Pele/citologia , Pele/efeitos da radiação , Envelhecimento da Pele/patologia
16.
Lasers Med Sci ; 35(5): 1205-1212, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32030556

RESUMO

The aim of this study was to assess the effects of IL-6 and IL-8 cytokines on human gingival fibroblasts (HGF) cultured in a 3-D model and the possible photobiomodulation (PBM) of such effects by low-level laser therapy. In complete culture medium (DMEM), HGF from a healthy patient were seeded in a type I collagen matrix inserted into 24-well plates. After 5 days of incubation, the cytokines were added or not to serum-free DMEM, which was applied to the cell-enriched matrices. Then, PBM was performed: three consecutive irradiations using LaserTable diode device (780 nm, 0.025 W) at 0.5 J/cm2 were delivered or not to the cells. Twenty-four hours after the last irradiation, cell viability and morphology, gene expression, and synthesis of inflammatory cytokines and growth factors were assessed. The histological evaluation demonstrated that, for all groups, matrices presented homogeneous distribution of cells with elongated morphology. However, numerous cytokine-exposed cells were rounded. IL-6 and IL-8 decreased cell viability, synthesis of VEGF, and gene expression of collagen type I. PBM enhanced cell density in the matrices and stimulated VEGF expression, even after IL-6 challenge. Reduced TNF-α synthesis occurred in those cells subjected to PBM. In conclusion, PBM can penetrate collagen matrix and stimulate HGF, highlighting the relevance of this research model for further phototherapy studies and in vitro biomodulation of the healing process.


Assuntos
Técnicas de Cultura de Células/métodos , Citocinas/metabolismo , Fibroblastos/patologia , Fibroblastos/efeitos da radiação , Gengiva/patologia , Inflamação/patologia , Terapia com Luz de Baixa Intensidade , Modelos Biológicos , Sobrevivência Celular/efeitos da radiação , Colágeno Tipo I/metabolismo , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Interleucina-1beta/biossíntese , Fator de Necrose Tumoral alfa/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossíntese , Cicatrização/efeitos da radiação
17.
Int J Mol Sci ; 21(3)2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-32041250

RESUMO

The present study aimed to explore the possible radioprotective effects of celastrol and relevant molecular mechanisms in an in vitro cell and in vivo mouse models exposed to gamma radiation. Human keratinocytes (HaCaT) and foreskin fibroblast (BJ) cells were exposed to gamma radiation of 20Gy, followed by treatment with celastrol for 24 h. Cell viability, reactive oxygen species (ROS), nitric oxide (NO) and glutathione (GSH) production, lipid peroxidation, DNA damage, inflammatory cytokine levels, and NF-κB pathway activation were examined. The survival rate, levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in blood, and p65 and phospho-p65 expression were also evaluated in mice after exposure to gamma radiation and celastrol treatment. The gamma irradiation of HaCaT cells induced decreased cell viability, but treatment with celastrol significantly blocked this cytotoxicity. Gamma irradiation also increased free radical production (e.g., ROS and NO), decreased the level of GSH, and enhanced oxidative DNA damage and lipid peroxidation in cells, which were effectively reversed by celastrol treatment. Moreover, inflammatory responses induced by gamma irradiation, as demonstrated by increased levels of IL-6, TNF-α, and IL-1ß, were also blocked by celastrol. The increased activity of NF-κB DNA binding following gamma radiation was significantly attenuated after celastrol treatment. In the irradiated mice, treatment with celastrol significantly improved overall survival rate, reduced the excessive inflammatory responses, and decreased NF-κB activity. As a NF-κB pathway blocker and antioxidant, celastrol may represent a promising pharmacological agent with protective effects against gamma irradiation-induced injury.


Assuntos
Prepúcio do Pênis/citologia , Raios gama/efeitos adversos , Queratinócitos/citologia , Protetores contra Radiação/farmacologia , Triterpenos/farmacologia , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Modelos Animais de Doenças , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/imunologia , Fibroblastos/efeitos da radiação , Prepúcio do Pênis/efeitos dos fármacos , Prepúcio do Pênis/imunologia , Prepúcio do Pênis/efeitos da radiação , Glutationa/efeitos dos fármacos , Glutationa/metabolismo , Glutationa/efeitos da radiação , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/imunologia , Queratinócitos/efeitos da radiação , Masculino , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Fator de Necrose Tumoral alfa/metabolismo
18.
Oxid Med Cell Longev ; 2020: 2576823, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32104530

RESUMO

UVA irradiation induced ROS-mediated photo damage to the human skin leading to coarseness, wrinkling, pigmentation, and cutaneous malignancies. We investigated the dermatoprotective efficacies of submicromolar concentrations of ergothioneine (EGT, 0.125-0.5 µM), which occurs naturally as a sulfur-containing amino acid, in the mechanisms in human skin fibroblast (HSF) cells. UVA-induced AP-1 (c-Fos and c-Jun) translocation was found to be inhibited by EGT treatments with the parallel inhibition of the collagenolytic matrix metalloproteinase- (MMP-) 1 activation and type I procollagen degradation. Moreover, EGT mitigated UVA-induced ROS generation. An increase in the amount of antioxidant genes (HO-1, NQO-1, and γ-GCLC) from EGT and were associated with upregulated Nrf2 expressions in a dose-dependent or time-dependent manner. We confirmed this from Nrf2 translocation and increased nuclear ARE promoter activity that underlie EGT dermatoprotective activities. Also, glutathione (GSH) levels (from γ-GCLC) were significantly increased. Moreover, we showed that mediated by ERK, JNK, and PKC, signaling cascades mediate Nrf2 translocation. We confirmed this phenomenon by the suppressed nuclear Nrf2 activation in cells that were treated with respective inhibitors (PD98059, SP600125, and GF109203X). However, antioxidant protein expressions were impaired in Nrf2 knockdown cells to confirm that ARE/Nrf2 pathways and the inhibition of AP-1 had significant roles in EGT-mediated protective effects. We can conclude that ergothioneine ameliorated UVA-induced skin aging and is a useful food supplement for skin care products.


Assuntos
Ergotioneína/farmacologia , Fibroblastos/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Transdução de Sinais/efeitos dos fármacos , Envelhecimento da Pele/efeitos dos fármacos , Fator de Transcrição AP-1/metabolismo , Antioxidantes/farmacologia , Linhagem Celular , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Glutationa/metabolismo , Heme Oxigenase-1/metabolismo , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , NAD(P)H Desidrogenase (Quinona)/metabolismo , Fator 2 Relacionado a NF-E2/genética , Pró-Colágeno/metabolismo , Proteína Quinase C/metabolismo , RNA Interferente Pequeno , Espécies Reativas de Oxigênio/metabolismo , Espécies Reativas de Oxigênio/efeitos da radiação , Transdução de Sinais/genética , Fator de Transcrição AP-1/antagonistas & inibidores , Raios Ultravioleta , Regulação para Cima/efeitos dos fármacos
19.
J Photochem Photobiol B ; 203: 111731, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31935633

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a progressive and chronic inflammatory disease with a poor prognosis and very few available treatment options. Low-level laser therapy (LLLT) has been gaining prominence as a new and effective anti-inflammatory and immunomodulatory agent. Can lung inflammation and the airway remodeling be regulated by LLLT in an experimental model of IPF in C57Bl/6 mice? The present study investigated if laser attenuates cellular migration to the lungs, the airway remodeling as well as pro-fibrotic cytokines secretion from type II pneumocytes and fibroblasts. Mice were irradiated (780 nm and 30 mW) and then euthanized fifteen days after bleomycin-induced lung fibrosis. Lung inflammation and airway remodeling were evaluated through leukocyte counting in bronchoalveolar lavage fluid (BALF) and analysis of collagen in lung, respectively. Inflammatory cells in blood were also measured. For in vitro assays, bleomycin-activated fibroblasts and type II pneumocytes were irradiated with laser. The pro- and anti-inflammatory cytokines level in BALF as well as cells supernatant were measured by ELISA, and the TGFß in lung was evaluated by flow cytometry. Lung histology was used to analyze collagen fibers around the airways. LLLT reduced both migration of inflammatory cells and deposition of collagen fibers in the lungs. In addition, LLLT downregulated pro-inflammatory cytokines and upregulated the IL-10 secretion from fibroblasts and pneumocytes. Laser therapy greatly reduced total lung TGFß. Systemically, LLLT also reduced the inflammatory cells counted in blood. There is no statistical difference in inflammatory parameters studied between mice of the basal group and the laser-treated mice. Data obtained indicate that laser effectively attenuates the lung inflammation, and the airway remodeling in experimental pulmonary fibrosis is driven to restore the balance between the pro- and anti-inflammatory cytokines in lung and inhibit the pro-fibrotic cytokines secretion from fibroblasts.


Assuntos
Remodelação das Vias Aéreas , Citocinas/metabolismo , Fibrose Pulmonar Idiopática/radioterapia , Lasers , Animais , Líquido da Lavagem Broncoalveolar/química , Células Cultivadas , Citocinas/análise , Modelos Animais de Doenças , Regulação para Baixo/efeitos da radiação , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/efeitos da radiação , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Fibrose Pulmonar Idiopática/patologia , Terapia a Laser , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Regulação para Cima/efeitos da radiação
20.
J Photochem Photobiol B ; 203: 111738, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31954290

RESUMO

This study aimed to compare the synthesis and secretion of VEGF-A, VEGF-C, VEGF-D, VEGFR1, VEGFR2, and FGF-2 between pulp fibroblasts from human primary teeth (HPF) and stem cell from human deciduous teeth (SHED) before and after photobiomodulation. HPF were obtained from explant technique and characterized by immunohistochemistry, while SHED were obtained from digestion technique and characterized by flow cytometry. HPF (control group) and SHED were plated, let to adhere, and put on serum starvation to synchronize the cell cycles prior to photobiomodulation. Then, both cell lineages were irradiated with 660-nm laser according to the following groups: 2.5 and 3.7 J/cm2. MTT and crystal violet assays respectively verified viability and proliferation. ELISA Multiplex Assay assessed the following proteins: VEGF-A, VEGF-C, VEGF-D, VEGFR1, VEGFR2, FGF-2, at 6, 12, and 24 h after photobiomodulation, in supernatant and lysate. Two-way ANOVA/Tukey test evaluated cell viability and proliferation, while angiogenic production and secretion values were analyzed by one-way ANOVA (P < .05). Statistically similar HPF and SHED viability and proliferation patterns occurred before and after photobiomodulation (P > .05). HPF exhibited statistically greater values of all angiogenic proteins than did SHED, at all study periods, except for FGF-2 (supernatant; 12 h); VEGFR1 (lysate; non-irradiated; 12 h); and VEGFR1 (lysate; non-irradiated; 24 h). Photobiomodulation changed the synthesis and secretion of angiogenic proteins by HPF. HPF produced and secreted greater values of all tested angiogenic proteins than did SHED before and after irradiation with both energy densities of 2.5 and 3.7 J/cm2.


Assuntos
Fibroblastos/efeitos da radiação , Lasers , Células-Tronco/efeitos da radiação , Linhagem da Célula/efeitos da radiação , Proliferação de Células/efeitos da radiação , Sobrevivência Celular/efeitos da radiação , Células Cultivadas , Fibroblastos/citologia , Fibroblastos/metabolismo , Humanos , Células-Tronco/citologia , Células-Tronco/metabolismo , Dente Decíduo/citologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...