RESUMO
Near-infrared (NIR) can penetrate the dermis. NIR is able to regulate cutaneous component cells and immune cells and shows significant anti-inflammatory therapeutic effects. However, the mechanisms of these effects are largely unknown. The purpose of this study is to elucidate NIR-induced molecular mechanisms on macrophages because macrophages play initial roles in directing immune responses by their M1 or M2 polarizations. Proteomic analysis revealed that NIR radiation enhanced the expression of mitochondrial respiratory gene citrate synthase. This increased citrate synthase expression was triggered by NIR-induced H3K4 hypermethylation on the citrate synthase gene promoter but not by heat, which led to macrophage M2 polarization and finally resulted in TGFß1 release from CD4+ cells. These cellular effects were validated in human primary macrophages and abdominal NIR-irradiated mouse experiments. In a phorbol 12-myristate 13-acetateâinduced inflammatory model on mouse ear, we confirmed that NIR irradiation induced significant anti-inflammatory effects through decreased M1 counts, reduced TNF-α, and increased CCL22 and/or TGFß1 levels.
Assuntos
Dermatite/terapia , Raios Infravermelhos/uso terapêutico , Macrófagos/imunologia , Fototerapia/métodos , Animais , Citrato (si)-Sintase/metabolismo , Dermatite/imunologia , Derme/citologia , Derme/imunologia , Derme/metabolismo , Derme/efeitos da radiação , Modelos Animais de Doenças , Feminino , Humanos , Ativação de Macrófagos/efeitos da radiação , Macrófagos/citologia , Macrófagos/metabolismo , Macrófagos/efeitos da radiação , Camundongos , Mitocôndrias/enzimologia , Mitocôndrias/efeitos da radiação , Cultura Primária de Células , Células THP-1RESUMO
To study the effect of photobiomodulation (PBM) on axon regeneration and secretion change of dorsal root ganglion (DRG) under oxidative stress after spinal cord injury (SCI), and further explore the effect of changes in DRG secretion caused by PBM on the polarization of macrophages. The PBM-DRG model was constructed to perform PBM on neurons under oxidative stress simulated in vitro. And the irradiation conditions were as follows: wavelength, 810 nm; power density, 2 mW/cm2; irradiation area, 4.5 cm2; and irradiation time, 440 s. Then resulted in an energy of 4 J (2 mW/cm2 × 4.5 cm2 × 440 s). About 100 µM H202 was added to the culture medium to simulate oxidative stress after SCI. An ROS (reactive oxygen species) assay kit was used to measure ROS contend in the DRG. The survival level of the neurons was measured using the CCK-8 method, and the axon regeneration of neurons was observed by using immunofluorescence. The secretion level of CCL2 from DRG was determined by RT-qPCR and ELISA. Further culturing macrophages of DRG-conditioned medium culture, the expression level of iNOS and Arg-1 in macrophages was assessed using Western blot analysis. The expression level of TNF-α and IL-1ß was determined by ELISA. After adding the neutralizing antibody of CCL2 to the DRG neuron-conditioned medium following PBM irradiation to culture macrophages to observe the effects on macrophage polarization and secretion. PBM could reduce ROS levels in neurons, increase neuronal survival under oxidative stress, and promote neuronal axon regeneration. In addition, PBM could also promote CCL2 secretion by DRG under oxidative stress. By constructing a DRG supernatant-M1 macrophage adoptive culture model, we found that the supernatant of DRG after PBM intervention could reduce the expression level of iNOS and the secretion of TNF-α and IL-1ß in M1 macrophages; at the same time, it could also up-regulate the expression of Arg-1, one of the markers of M2 macrophages. Furthermore, these effects could be prevented by the addition of neutralizing antibodies of CCL2. PBM could promote survival and axonal regeneration of DRG under SCI oxidative stress, increase the secretion level of CCL2 by DRG, and this change can reduce the polarization of macrophages to M1, further indicating that PBM could promote spinal cord injury repair.
Assuntos
Axônios/metabolismo , Quimiocina CCL2/metabolismo , Macrófagos/citologia , Estresse Oxidativo , Fototerapia/métodos , Traumatismos da Medula Espinal/terapia , Regeneração da Medula Espinal , Animais , Axônios/efeitos da radiação , Diferenciação Celular , Células Cultivadas , Quimiocina CCL2/genética , Feminino , Gânglios Espinais/citologia , Gânglios Espinais/metabolismo , Gânglios Espinais/fisiologia , Interleucina-1beta/metabolismo , Luz , Macrófagos/imunologia , Macrófagos/efeitos da radiação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Several reports have suggested that photobiomodulation, owing to its analgesic, anti-inflammatory, and healing effects, may be an effective therapeutic option for local effects of snakebites when the availability and accessibility of conventional serum therapy are inefficient and far from medical care centers. Although there have been studies that demonstrate the application of photobiomodulation in the treatment of local adverse events due to snakebites from snakes of the genus Bothrops, its role in the activation of leukocytes, particularly macrophages, has not been evaluated. Here, we assessed the effect of light-emitting diode (LED) treatment on macrophage activation induced by B. jararacussu venom (BjV). LED treatment caused an increase in the viability of macrophages incubated with BjV. This treatment reduced reactive oxygen species (ROS) and nitric oxide (NO) production by macrophages after incubation with BjV. However, LED treatment did not interfere with IL-1ß and IL-10 production by macrophages after incubation with BjV. In conclusion, this study showed that LED treatment has the potential to be used in combination with conventional serum therapy to prevent or minimize the progression of local to severe symptoms after Bothrops envenomation.
Assuntos
Bothrops , Venenos de Crotalídeos/toxicidade , Terapia com Luz de Baixa Intensidade/instrumentação , Macrófagos/efeitos da radiação , Semicondutores , Mordeduras de Serpentes/imunologia , Mordeduras de Serpentes/radioterapia , Animais , Sobrevivência Celular/efeitos dos fármacos , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Espaço Intracelular/efeitos da radiação , Macrófagos/imunologia , Masculino , Camundongos , Óxido Nítrico/biossíntese , Espécies Reativas de Oxigênio/metabolismo , Mordeduras de Serpentes/metabolismo , Mordeduras de Serpentes/patologia , Superóxidos/metabolismoRESUMO
During spaceflight, the homeostasis of the living body is threatened with cosmic environment including microgravity and irradiation. Traditional Chinese medicine could ameliorate the internal imbalance during spaceflight, but its mechanism is still unclear. In this article, we compared the difference of neuroendocrine-immune balance between simulated microgravity (S) and simulated microgravity and irradiation (SAI) environment. We also observed the antagonistic effect of SAI using a traditional Chinese medicine formula (TCMF). Wistar rats were, respectively, exposed under S using tail suspending and SAI using tail suspending and 60Co-gama irradiation exposure. The SAI rats were intervened with TCMF. The changes of hypothalamic-pituitary-adrenal (HPA) axis, splenic T-cell, celiac macrophages, and related cytokines were observed after 21 days. Compared with the normal group, the hyperfunction of HPA axis and celiac macrophages, as well as the hypofunction of splenic T-cells, was observed in both the S and SAI group. Compared with the S group, the levels of plasmatic corticotropin-releasing hormone (CRH), macrophage activity, and serous interleukin-6 (IL-6) in the SAI group were significantly reduced. The dysfunctional targets were mostly reversed in the TCMF group. Both S and SAI could lead to NEI imbalance. Irradiation could aggravate the negative feedback inhibition of HPA axis and macrophages caused by S. TCMF could ameliorate the NEI dysfunction caused by SAI.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Sistema Imunitário , Sistemas Neurossecretores , Ausência de Peso , Animais , Corticosterona/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Citocinas/metabolismo , Raios gama , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/efeitos da radiação , Sistema Imunitário/efeitos dos fármacos , Sistema Imunitário/efeitos da radiação , Macrófagos/efeitos dos fármacos , Macrófagos/efeitos da radiação , Masculino , Medicina Tradicional Chinesa , Modelos Biológicos , Sistemas Neurossecretores/efeitos dos fármacos , Sistemas Neurossecretores/efeitos da radiação , Ratos , Ratos Wistar , Linfócitos T/efeitos dos fármacos , Linfócitos T/efeitos da radiaçãoRESUMO
We determined the impact of Photobiomodulation (PBM) and metformin administration alone and combined on the inflammation and proliferation steps of wound healing of incisions in type two diabetes mellitus (T2DM) rats. 40 rats were divided into 4 groups (nâ¯=â¯10 each group). A non-genetic model of T2DM was induced in all rats, and an incision was made on each rat. There were 4 groups as follows: Group 1 was control group. Group 2 received PBM alone (890â¯nm, 80â¯Hz, 0.324â¯J/cm2, daily). Group 3 received metformin alone (50â¯mg/kg, i.p., daily) and the fourth group received combination of PBMâ¯+â¯metformin. At inflammation (day 4) and proliferation (day 7) steps, tensiometerical, stereological, and immunohistochemical examinations were performed. PBM and PBMâ¯+â¯metformin treatments significantly increased wound strength at inflammation and proliferation steps of wound healing respectively. PBM, metformin, and PBMâ¯+â¯metformin groups significantly decreased inflammatory cells at inflammation and proliferation steps of wound healing. PBM, metformin, and PBMâ¯+â¯metformin groups significantly improved granulation tissue formation by increasing fibroblasts, and new blood vessel formation at inflammation and proliferation steps of wound healing. Metformin significantly increased M2 macrophages than other treatment groups at inflammation and proliferation steps of wound healing. Simultaneously, PBM significantly decreased M2 macrophages than control group. We concluded PBM and PBMâ¯+â¯metformin treatments significantly hastened repair at the inflammation and proliferation steps of repairing skin injury in a non-genetic model of T2 DM. PBMâ¯+â¯metformin showed a synergistic impact. There were not a positive relation between M2 macrophage number and wound strength in the studied groups. The details of the molecular mechanisms of PBM, and PBMâ¯+â¯metformin treatments of repairing wounds in animals, and treatment of DFUs of patients with T2 DM should be elucidated by further research.
Assuntos
Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Tipo 2/terapia , Terapia com Luz de Baixa Intensidade , Metformina/farmacologia , Cicatrização , Animais , Glicemia/metabolismo , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Terapia Combinada , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/patologia , Tecido de Granulação/efeitos dos fármacos , Tecido de Granulação/patologia , Tecido de Granulação/efeitos da radiação , Inflamação/patologia , Macrófagos/efeitos dos fármacos , Macrófagos/efeitos da radiação , Masculino , Ratos Wistar , Cicatrização/efeitos dos fármacos , Cicatrização/efeitos da radiaçãoRESUMO
Ultraviolet B (UVB) induces inflammation and causes skin aging. The signs of skin aging, such as wrinkles, discolored spots, loss of skin moisture, and disruption of the skin barrier, are mostly caused by inflammatory signaling among various skin layers. The cells on the outermost surface of the skin are keratinocytes; these cells protect the skin against environmental stress and play an important role in immunomodulation by secreting cytokines in response to environmental stress. In the present study, we found that UVB activates STAT1 to mediate inflammatory signaling, yet STAT1 (S272) and STAT (Y702) shows different responses against UVB exposure. Anhua drak tea is a post-fermented dark tea produced in Anhua and Xinhua country in Hunan province of China. Treatment with 2S,3R-6-methoxycarbonylgallocatechin (MCGE), an epigallocatechin gallate derivative isolated from black tea (Anhua dark tea), effectively suppresses STAT1 activation and inflammatory cytokines, and activates Nrf2 pathway to protect cells from reactive oxygen species production in UVB exposed keratinocyte cells (HaCaT). Interestingly, the effects of MCGE were independent on MAPK signaling pathway. Moreover, MCGE regulates inflammatory cytokines in monocyte-keratinocyte (THP-1, HaCaT) co-culture and macrophage differentiation models. These results suggest that MCGE potentially can be used as a photoprotective agent against UVB-induced inflammatory responses.
Assuntos
Catequina/análogos & derivados , Catequina/farmacologia , Protetores contra Radiação/farmacologia , Transdução de Sinais/efeitos dos fármacos , Chá/química , Raios Ultravioleta , Sítios de Ligação , Catequina/química , Catequina/isolamento & purificação , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/efeitos da radiação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Técnicas de Cocultura , Citocinas/metabolismo , Humanos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Queratinócitos/efeitos da radiação , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/efeitos da radiação , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Estrutura Terciária de Proteína , Protetores contra Radiação/química , Protetores contra Radiação/isolamento & purificação , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição STAT1/química , Fator de Transcrição STAT1/metabolismo , Transdução de Sinais/efeitos da radiação , Chá/metabolismoRESUMO
In response to stimuli in the microenvironment, macrophages adopt either the M1 or M2 phenotype to coordinate the tissue repair process. Photobiomodulation (PBM) plays an important role in the modulation of acute inflammation, including cellular influx, macrophage polarization, and the release of inflammatory mediators. The aim of the present study was to evaluate the effects of red and infrared PBM on the mRNA expression of cytokines and chemokines in macrophages polarized to the M1 and M2 phenotypes. J774 macrophages activated to induce M1 (lipopolysaccharide + interferon gamma) or M2 (interleukin-4) phenotypes were irradiated with red or infrared PBM (1 J). After 4 and 24 h, gene expression was analyzed by qPCR. PBM at 660 nm decreased the mRNA expression of CCL3, CXCL2, and TNF-α in M1 macrophages and CXCL2 in M2 macrophages 4 h after irradiation. Similarly, PBM at 780 nm decreased mRNA expression levels of CCL3 and IL-6 by M1 macrophages 24 h after irradiation. Moreover, PBM at 780 nm increased the mRNA expression of TGFß1 4 h after irradiation and decreased the expression of this gene after 24 h in M2 macrophages. Although red and infrared PBM were able to modulate and reduce M1/M2a-related markers, infrared laser irradiation promoted a temporal increase in the expression of TGFß1 in M2 macrophages. Thus, depending on the time PBM is used on injured tissue, different parameters can promote optimal results by modulating specific macrophage phenotypes.
Assuntos
Mediadores da Inflamação/metabolismo , Inflamação/metabolismo , Raios Infravermelhos , Lasers , Terapia com Luz de Baixa Intensidade , Macrófagos/metabolismo , Macrófagos/efeitos da radiação , Animais , Biomarcadores/metabolismo , Linhagem Celular , Regulação da Expressão Gênica/efeitos da radiação , Inflamação/patologia , Ativação de Macrófagos/efeitos dos fármacos , Camundongos Endogâmicos BALB C , FenótipoRESUMO
Spinal cord injury (SCI) stimulates reactive astrogliosis and the infiltration of macrophages, which interact with each other at the injured area. We previously found Photobiomodulation (PBM) significantly decreases the number of M1 macrophages at the injured area of SCI. But the exact nature of the astrocyte response following PBM and relationship with the macrophage have not been explored in detail. In this study, a BALB/c mice model with standardized bilateral spinal cord compression and a macrophage-astrocyte co-culture model were applied to study effects of PBM on astrocytes. Results showed that PBM inhibit the expression of the astrocyte markers glial fibrillary acidic protein (GFAP) and the secretion of chondroitin sulfate proteoglycans (CSPG) in the para-epicenter area, decrease the number of M1 macrophage in vivo. The in vitro experiments indicated M1 macrophages promote the cell viability of astrocytes and the expression of CSPG. However, PBM significantly inhibited the expression of GFAP, decreased activation of astrocyte, and downregulated the expression of CSPG by regulating M1 macrophages. These results demonstrate that PBM may regulate the interaction between macrophages and astrocytes after spinal cord injury, which inhibited the formation of glial scar.
Assuntos
Astrócitos/efeitos da radiação , Polaridade Celular/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Macrófagos/efeitos da radiação , Animais , Astrócitos/efeitos dos fármacos , Polaridade Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Proteoglicanas de Sulfatos de Condroitina/metabolismo , Meios de Cultivo Condicionados/farmacologia , Feminino , Proteína Glial Fibrilar Ácida/metabolismo , Macrófagos/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Atividade Motora/efeitos dos fármacos , Atividade Motora/efeitos da radiação , Fosforilação/efeitos dos fármacos , Fosforilação/efeitos da radiação , Recuperação de Função Fisiológica/efeitos dos fármacos , Fator de Transcrição STAT3/metabolismo , Traumatismos da Medula Espinal/fisiopatologia , Traumatismos da Medula Espinal/radioterapiaRESUMO
Macrophages play key roles in the secondary injury stage of spinal cord injury (SCI). M1 macrophages occupy the lesion area and secrete high levels of inflammatory factors that hinder lesion repair, and M2 macrophages can secrete neurotrophic factors and promote axonal regeneration. The regulation of macrophage secretion after SCI is critical for injury repair. Low-level laser therapy (810-nm) (LLLT) can boost functional rehabilitation in rats after SCI; however, the mechanisms remain unclear. To explore this issue, we established an in vitro model of low-level laser irradiation of M1 macrophages, and the effects of LLLT on M1 macrophage polarization and neurotrophic factor secretion and the related mechanisms were investigated. The results showed that LLLT irradiation decreased the expression of M1 macrophage-specific markers, and increased the expression of M2 macrophage-specific markers. Through forward and reverse experiments, we verified that LLLT can promote the secretion of various neurotrophic factors by activating the PKA-CREB pathway in macrophages and finally promote the regeneration of axons. Accordingly, LLLT may be an effective therapeutic approach for SCI with clinical application prospects.
Assuntos
Axônios/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Terapia com Luz de Baixa Intensidade , Macrófagos/metabolismo , Macrófagos/efeitos da radiação , Fatores de Crescimento Neural/metabolismo , Regeneração Nervosa , Animais , Axônios/efeitos dos fármacos , Axônios/efeitos da radiação , Meios de Cultivo Condicionados/farmacologia , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Feminino , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/metabolismo , Isoquinolinas/farmacologia , Macrófagos/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Fatores de Crescimento Neural/genética , Regeneração Nervosa/efeitos dos fármacos , Regeneração Nervosa/efeitos da radiação , Inibidores de Proteínas Quinases/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sulfonamidas/farmacologiaRESUMO
The light-emitting diode (LED) is considered a therapeutic tool due to its anti-inflammatory, analgesic, and wound-healing effects, which occur through angiogenesis, decrease in IL-1ß and IL-6 secretion, and acceleration of the cicatricial process. Snakebites are an important public health problem in tropical regions of the world. LED treatment is a therapeutic tool associated with serum therapy used to minimize the local effects of snakebites, including decrease in creatine kinase (CK) and lactate dehydrogenase (LDH) concentrations, myonecrosis, and inflammatory and haemorrhagic responses. In this study, we analysed the photobiomodulation effect of LED on the activation of murine macrophages induced by BthTX-I or BthTX-II isolated from Bothrops jararacussu venom. Photobiomodulation caused an increase in mitochondrial metabolism and a considerable decrease in cytotoxicity in murine macrophages. Moreover, it induced a decrease in reactive oxygen species and nitrogen liberation. However, photobiomodulation caused an increase in macrophage phagocytic capacity and lipid droplet formation. The results of this study corroborated with those of others in an unprecedented way and provide a better understanding of the mechanism of action of photobiomodulation, besides offering a coadjuvant action treatment for the local effects of snakebites, not achieved with serum therapy alone.
Assuntos
Venenos de Crotalídeos/toxicidade , Fosfolipases A2 do Grupo II/toxicidade , Terapia com Luz de Baixa Intensidade , Macrófagos/efeitos dos fármacos , Macrófagos/efeitos da radiação , Animais , Bothrops , Masculino , Camundongos , Mitocôndrias/metabolismo , Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Inflammatory macrophage (Mφ)-mediated atherosclerosis is a leading cause of mortality and morbidity worldwide. Photothermal therapy (PTT) has been demonstrated as an efficient strategy in killing target cells, and its application in the treatment of inflammation in atherosclerosis is developing. However, the choice of nanomaterials, mechanisms, and side effects are seldom considered. In this study, semiconductor nanomaterials, that is, MoO2 nanoclusters, were synthesized and used for the first time in PTT for inflammatory Mφ-mediated atherosclerosis. Based on cell differential phagocytosis, the optimum amount of MoO2 and treatment time were selected to exert the maximum ablation effect on Mφ and minimal damage on endothelial cells without requiring additional target or selective groups. Moreover, MoO2-based PTT shows an excellent therapeutic effect on atherosclerosis by eliminating Mφ in animal models, with no significant side effects observed. This study explores a new method of nanotechnology and pharmaceutical development by using and optimizing cost-effective metal oxide nanostructures in the treatment of atherosclerosis and motivates further research on minimizing the side effects of related materials.
Assuntos
Aterosclerose/terapia , Raios Infravermelhos , Macrófagos/efeitos da radiação , Fagocitose/efeitos da radiação , Animais , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Aterosclerose/imunologia , Aterosclerose/patologia , Células da Medula Óssea/citologia , Artérias Carótidas/efeitos dos fármacos , Artérias Carótidas/patologia , Artérias Carótidas/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Modelos Animais de Doenças , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Humanos , Macrófagos/citologia , Macrófagos/imunologia , Masculino , Nanopartículas Metálicas/química , Nanopartículas Metálicas/uso terapêutico , Nanopartículas Metálicas/toxicidade , Camundongos , Camundongos Knockout , Molibdênio/química , Molibdênio/farmacocinética , Óxidos/química , Óxidos/farmacocinética , Fototerapia , Semicondutores , Distribuição TecidualRESUMO
OBJECTIVE: Tumor-treating fields are currently used to successfully treat various cancers; however, the specific pathways associated with its efficacy remain unknown in the immune responses. Here, we evaluated tumor-treating fields-mediated initiation of the macrophage-specific immune response. MATERIALS AND METHODS: We subjected RAW 264.7 mouse macrophages to clinically relevant levels of tumor-treating fields (0.9 V/cm, 150 kHz) and evaluated alterations in cytokine expression and release, as well as cell viability. Additionally, we investigated the status of immunomodulatory pathways to determine their roles in tumor-treating fields-mediated immune activation. RESULTS AND DISCUSSION: Our results indicated that tumor-treating fields treatment at 0.9 V/cm decreased cell viability and increased cytokine messenger RNA/protein levels, as well as levels of nitric oxide and reactive oxygen species, relative to controls. The levels of tumor necrosis factor α, interleukin 1ß, and interleukin 6 were markedly increased in tumor-treating fields-treated RAW 264.7 cells cocultured with 4T1 murine mammary carcinoma cells compared with those in 4T1 or RAW 264.7 cells with or without tumor-treating fields treatment. Moreover, the viability of 4T1 cells treated with the conditioned medium of tumor-treating fields-stimulated RAW 264.7 cells decreased, indicating that macrophage activation by tumor-treating fields effectively killed the tumor cells. Moreover, tumor-treating fields treatment activated the nuclear factor κB and mitogen-activated protein kinase pathways involved in immunomodulatory signaling. CONCLUSION: These results provide critical insights into the mechanisms through which tumor-treating fields affect macrophage-specific immune responses and the efficacy of this method for cancer treatment.
Assuntos
Antígenos de Histocompatibilidade Classe II/imunologia , Ativação de Macrófagos/imunologia , Magnetoterapia , Neoplasias/radioterapia , Animais , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe II/efeitos da radiação , Humanos , Sistema de Sinalização das MAP Quinases/efeitos da radiação , Ativação de Macrófagos/genética , Ativação de Macrófagos/efeitos da radiação , Macrófagos/imunologia , Macrófagos/efeitos da radiação , Camundongos , NF-kappa B/genética , Neoplasias/imunologia , Neoplasias/patologia , Células RAW 264.7 , Transdução de Sinais/imunologia , Transdução de Sinais/efeitos da radiaçãoRESUMO
We examined the effects of photobiomodulation (PBM) independently and combined with curcumin on stereological parameters and basic fibroblast growth factor (bFGF), hypoxia-inducible factor-1α (HIF-1α), and stromal cell-derived factor-1α (SDF-1α) gene expressions in an excisional wound model of rats with type one diabetes mellitus (T1DM). T1DM was induced by an injection of streptozotocin (STZ) in each of the 90 male Wistar rats. One round excision was generated in the skin on the back of each of the 108 rats. The rats were divided into six groups (n = 18 per group): control (diabetic), untreated group; vehicle (diabetic) group, which received sesame oil; PBM (diabetic) group; curcumin (diabetic) group; PBM + curcumin (diabetic) group; and a healthy control group. On days 4, 7, and 15, we conducted both stereological and quantitative real-time PCR (qRT-PCR) analyses. The PBM and PBM + curcumin groups had significantly better inflammatory response modulation in terms of macrophages (P < .01), neutrophils (P < .001), and increased fibroblast values compared with the other groups at day 4 (P < .001), day 7 (P < .01), and day 15 (P < .001). PBM treatment resulted in increased bFGF gene expression on days 4 (P < .001) and 7 (P < .001), and SDF-1α gene expression on day 4 (P < .001). The curcumin group had increased bFGF (P < .001) expression on day 4. Both the PBM and PBM + curcumin groups significantly increased wound healing by modulation of the inflammatory response, and increased fibroblast values and angiogenesis. The PBM group increased bFGF and SDF-1α according to stereological and gene expression analyses compared with the other groups. The PBM and PBM + curcumin groups significantly increased the skin injury repair process to more rapidly reach the proliferation phase of the wound healing in T1DM rats.
Assuntos
Curcumina/uso terapêutico , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/radioterapia , Diabetes Mellitus Tipo 1/tratamento farmacológico , Diabetes Mellitus Tipo 1/radioterapia , Regulação da Expressão Gênica , Terapia com Luz de Baixa Intensidade , Cicatrização , Análise de Variância , Animais , Curcumina/farmacologia , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Tipo 1/genética , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibroblastos/efeitos da radiação , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos da radiação , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Macrófagos/efeitos da radiação , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/efeitos da radiação , Ratos Wistar , Cicatrização/efeitos dos fármacos , Cicatrização/genética , Cicatrização/efeitos da radiaçãoRESUMO
Objective: The aim of our study was to quantify the effect of doses delivered by a He:Ne laser on individual macrophage kinetics, tissue oxidative stress, and wound closure using real-time in vivo imaging. Background: Photobiomodulation has been reported to reduce tissue inflammation and accelerate wound closure; however, precise parameters of laser settings to optimize macrophage behavior have not been established. We hypothesized that quantitative and real-time in vivo imaging could identify optimal fluence for macrophage migration, reduction of reactive oxygen species, and acceleration of wound closure. Methods: Larval zebrafish Tg(mpeg-dendra2) were loaded with dihydroethidium for oxidative stress detection. Fish were caudal fin injured, treated with 635 nm continuous 5 mW He:Ne laser irradiation at 3, 9, or 18 J/cm2 and time-lapsed imaged within the first 120 min postinjury. Images taken 1 and 24-h postinjury were compared for percentage wound closure. Results: A fluence of 3 J/cm2 demonstrated significant increases in macrophage migration speed, fewer stops along the way, and greatest directed migration toward the wound. These findings were associated with a significant reduction in wound content reactive oxygen species when compared with control wounded fins. Both 3 and 9 J/cm2 significantly accelerated wound closure when compared with nonirradiated control fish. Conclusions: Wound macrophage activity could be manipulated by applied fluence, leading to reduced levels of wound reactive oxygen species and accelerated wound closure. The zebrafish model provides a means to quantitatively compare wound macrophage behavior in response to a variety of laser treatment parameters in real time.
Assuntos
Terapia com Luz de Baixa Intensidade/métodos , Macrófagos/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Cicatrização/efeitos da radiação , Animais , Movimento Celular/efeitos da radiação , Cinética , Microscopia de Fluorescência , Peixe-ZebraRESUMO
In this study, a series of subphthalocyanines (SubPcs) derivatives were synthesized to generate unique immunomodulatory molecules that can be activated through photo-induction. Immunomodulatory agents have a great potential in medicine to manipulate the immune system according to our needs and prevent disease symptoms. Inflammation is one of these symptoms and macrophages play a crucial role in the generation of inflammatory responses. Being able to control the activity of these agents through photo-induction enables the fine tuning on their activities in a location specific and non-invasive manner with possibly minor side effects. Mammalian macrophages' pro-inflammatory activity was examined in the presence of our compounds as well as LPS as a danger mimic. These compounds exerted photo-induced anti-inflammatory activities on the macrophages. Number of Cl atoms was a defining factor in their photo-induced anti-inflammatory immunomodulatory efficiencies.
Assuntos
Citocinas/efeitos da radiação , Mediadores da Inflamação/efeitos da radiação , Macrófagos/efeitos da radiação , Fármacos Fotossensibilizantes/farmacologia , Fototerapia/métodos , Animais , Linhagem Celular , Compostos Clorados/farmacologia , Indóis/farmacologia , Isoindóis , Lasers de Gás/uso terapêutico , CamundongosRESUMO
Intervertebral disc (IVD) degeneration is associated with imbalances between catabolic and anabolic responses, regulated by extracellular matrix (ECM)-modifying enzymes such as matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors of metalloproteinases (TIMPs). Potential contributing factors, such as interleukin (IL)-1ß and tumor necrosis factor (TNF)-α, derived from infiltrated, activated macrophages within IVD tissues, can trigger abnormal production of ECM-modifying enzymes and progression of IVD degeneration. Novel therapies for regulating ECM-modifying enzymes can prevent or ameliorate IVD degeneration. Photobiomodulation (PBM), known to regulate wound repair, exhibits regenerative potential by modulating biological molecules. This study examined the effects of PBM, administered at various wavelengths (630, 525, and 465 nm) and energy densities (16, 32, and 64 J/cm2), on the production of ECM-modifying enzymes in replicated degenerative IVD. Our results showed that PBM selectively inhibited the production of ECM-modifying enzymes in a dose- and wavelength-dependent manner, suggesting that it could be a novel tool for treating symptomatic IVD degeneration.
Assuntos
Matriz Extracelular/enzimologia , Degeneração do Disco Intervertebral/terapia , Terapia com Luz de Baixa Intensidade , Núcleo Pulposo/enzimologia , Progressão da Doença , Matriz Extracelular/efeitos da radiação , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Interleucina-1beta/genética , Disco Intervertebral/enzimologia , Disco Intervertebral/patologia , Disco Intervertebral/efeitos da radiação , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Macrófagos/patologia , Macrófagos/efeitos da radiação , Metaloproteinases da Matriz/genética , Metaloproteinases da Matriz/efeitos da radiação , Núcleo Pulposo/patologia , Núcleo Pulposo/efeitos da radiação , Cultura Primária de Células , Inibidores Teciduais de Metaloproteinases/genética , Inibidores Teciduais de Metaloproteinases/efeitos da radiação , Fator de Necrose Tumoral alfa/genéticaRESUMO
Macrophages play a very important role in the conduction of several regenerative processes mainly due to their plasticity and multiple functions. In the muscle repair process, while M1 macrophages regulate the inflammatory and proliferative phases, M2 (anti-inflammatory) macrophages direct the differentiation and remodelling phases, leading to tissue regeneration. The aim of this study was to evaluate the effect of red and near infrared (NIR) photobiomodulation (PBM) on macrophage phenotypes and correlate these findings with the repair process following acute muscle injury. Wistar rats were divided into 4 groups: control; muscle injury; muscle injury + red PBM; and muscle injury + NIR PBM. After 2, 4 and 7 days, the tibialis anterior muscle was processed for analysis. Macrophages phenotypic profile was evaluated by immunohistochemistry and correlated with the different stages of the skeletal muscle repair by the qualitative and quantitative morphological analysis as well as by the evaluation of IL-6, TNF-α and TGF-ß mRNA expression. Photobiomodulation at both wavelengths was able to decrease the number of CD68+ (M1) macrophages 2 days after muscle injury and increase the number of CD163+ (M2) macrophages 7 days after injury. However, only NIR treatment was able to increase the number of CD206+ M2 macrophages (Day 2) and TGF-ß mRNA expression (Day 2, 4 and 7), favouring the repair process more expressivelly. Treatment with PBM was able to modulate the inflammation phase, optimize the transition from the inflammatory to the regeneration phase (mainly with NIR light) and improve the final step of regeneration, enhancing tissue repair.
Assuntos
Terapia com Luz de Baixa Intensidade , Desenvolvimento Muscular/efeitos da radiação , Músculos/efeitos da radiação , Regeneração/efeitos da radiação , Animais , Antígenos CD/genética , Antígenos de Diferenciação Mielomonocítica/genética , Diferenciação Celular/efeitos da radiação , Humanos , Macrófagos/patologia , Macrófagos/efeitos da radiação , Músculo Esquelético/crescimento & desenvolvimento , Músculo Esquelético/lesões , Músculo Esquelético/efeitos da radiação , Músculos/lesões , Músculos/patologia , Ratos , Receptores de Superfície Celular/genética , Cicatrização/fisiologia , Cicatrização/efeitos da radiaçãoRESUMO
Activated macrophages have been known to play pivotal roles in the pathogenesis of rheumatoid arthritis (RA). 18F-FEDAC (N-benzyl-N-methyl-2-[7,8-dihydro-7-(2-18F-fluoroethyl)-8-oxo-2-phenyl-9H-purin-9-yl]acetamide) is a radiolabeled ligand for the 18-kDa translocator protein (TSPO), which is abundant in activated macrophages. We evaluated the feasibility of using 18F-FEDAC in a murine RA model. Methods: RAW 264.7 mouse macrophages were activated by lipopolysaccharide. TSPO expression levels in activated and inactivated macrophages were measured by quantitative polymerase chain reaction and Western blotting. The cellular uptake and specific binding of 18F-FEDAC were measured using a γ-counter. For the in vivo study, collagen-induced arthritis (CIA) was developed in DBA/1 mice, and the clinical score for arthritis was measured regularly. 18F-FEDAC and 18F-FDG PET images were acquired on days 23 and 37 after the first immunization. Histologic examinations were performed to evaluate macrophages and TSPO expression. Results: We found increased TSPO messenger RNA and protein expression in activated macrophages. Uptake of 18F-FEDAC in activated macrophages was higher than that in nonactivated cells and was successfully blocked by the competitor, PK11195. In CIA mice, joint swelling was apparent on day 26 after the first immunization, and the condition worsened by day 37. 18F-FEDAC uptake by arthritic joints increased early on (day 23), whereas 18F-FDG uptake did not. However, 18F-FDG uptake by arthritic joints markedly increased at later stages (day 37) to a higher level than 18F-FEDAC uptake. The 18F-FEDAC uptake correlated weakly with summed severity score (P = 0.019, r = 0.313), whereas the 18F-FDG uptake correlated strongly with summed severity score (P < 0.001, r = 0.897). Histologic sections of arthritic joints demonstrated an influx of macrophages compared with that in normal joints. Conclusion:18F-FEDAC enabled the visualization of active inflammation sites in arthritic joints in a CIA model by targeting TSPO expression in activated macrophages. The results suggest the potential usefulness of 18F-FEDAC imaging in the early phase of RA.
Assuntos
Acetamidas/química , Artrite/metabolismo , Colágeno/química , Fluordesoxiglucose F18/química , Macrófagos/efeitos da radiação , Purinas/química , Animais , Artrite Reumatoide/metabolismo , Linhagem Celular , Modelos Animais de Doenças , Radioisótopos de Flúor , Regulação Neoplásica da Expressão Gênica , Ligantes , Lipopolissacarídeos/química , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos DBA , Tomografia por Emissão de Pósitrons , Células RAW 264.7 , Compostos RadiofarmacêuticosRESUMO
Müller cells, the supporting cells of the retina, play a key role in responding to retinal stress by releasing chemokines, including CCL2, to recruit microglia and macrophages (MG/MΦ) into the damaged retina. Photobiomodulation (PBM) with 670 nm light has been shown to reduce inflammation in models of retinal degeneration. In this study, we aimed to investigate whether 670 nm light had an effect on Müller cell-initiated inflammation under retinal photo-oxidative damage (PD) in vivo and in vitro. Sprague-Dawley rats were pre-treated with 670 nm light (9J/cm2) once daily over 5 days prior to PD. The expression of inflammatory genes including CCL2 and IL-1ß was analysed in retinas. In vitro, primary Müller cells dissociated from neonatal rat retinas were co-cultured with 661W photoreceptor cells. Co-cultures were exposed to PD, followed by 670 nm light treatment to the Müller cells only, and Müller cell stress and inflammation were assessed. Primary MG/MΦ were incubated with supernatant from the co-cultures, and collected for analysis of inflammatory activation. To further understand the mechanism of 670 nm light, the expression of COX5a and mitochondrial membrane potential (ΔΨm) were measured in Müller cells. Following PD, 670 nm light-treated Müller cells had a reduced inflammatory activation, with lower levels of CCL2, IL-1ß and IL-6. Supernatant from 670 nm light-treated co-cultures reduced activation of primary MG/MΦ, and lowered the expression of pro-inflammatory cytokines, compared to untreated PD controls. Additionally, 670 nm light-treated Müller cells had an increased expression of COX5a and an elevated ΔΨm following PD, suggesting that retrograde signaling plays a role in the effects of 670 nm light on Müller cell gene expression. Our data indicates that 670 nm light reduces Müller cell-mediated retinal inflammation, and offers a potential cellular mechanism for 670 nm light therapy in regulating inflammation associated with retinal degenerations.
Assuntos
Células Ependimogliais/efeitos da radiação , Macrófagos/efeitos da radiação , Microglia/efeitos da radiação , Degeneração Retiniana/radioterapia , Animais , Quimiocinas/metabolismo , Grupo dos Citocromos c/metabolismo , Modelos Animais de Doenças , Células Ependimogliais/fisiologia , Interleucinas/metabolismo , Potencial da Membrana Mitocondrial/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Ratos , Ratos Sprague-Dawley , Degeneração Retiniana/metabolismoRESUMO
Since asthma is a multifactorial disease where treatment sometimes is not effective, new therapies that improve the respiratory discomfort of patients are of great importance. Phototherapy as Light-emitting diode (LED) has emerged as a treatment that presents good results for diseases that are characterized by inflammation. Thus, our objective was to investigate the effects of LED on lung inflammation, by an evaluation of lung cell infiltration, mucus secretion, oedema, and the production of cytokines. Male Balb/c mice were or not sensitized and challenged with ovalbumin (OVA) and treated or not with LED therapy (1â h and 4â h after each OVA challenge). Twenty-four hours after the last OVA challenge, analyzes were performed. Our results showed that LED treatment in asthmatic mice reduced the lung cell infiltration, the mucus production, the oedema, and the tracheal's contractile response. It also increased the IL-10 and the IFN-gamma levels. The effects of LED treatment on lung inflammation may be modulated by IL-10, IFN-gamma, and by mast cells. This study may provide important information about the effects of LED, and in addition, it may open the possibility of a new approach for the treatment of asthma.