RESUMO
Tubulointerstitial fibrosis is a common pathological change in end-stage renal disease. However, limited treatment methods are developed, and unexplained potential mechanisms of renal diseases are urgent problems to be solved. In the present research, we first elucidated the role of podocarpusflavone (POD), a biflavone compound, in unilateral ureteral obstruction (UUO) in rodent model which is characterized by inflammation and fibrosis. The changes in histology and immunohistochemistry were observed that POD exerted renoprotective effects by retarding the infiltration of macrophage and aberrant deposition of É-SMA, Col1a1, and fibronectin. Consistent with in vivo assay, POD treatment also ameliorated the process of fibrosis in TGF-ß1-stimulated renal tubular epithelial cells and inflammation in LPS-induced RAW264.7 cells in vitro. In terms of mechanism, our results showed that treatment with POD inhibited the aggravated activation of Fyn in the UUO group, and weakened the level of phosphorylation of Stat3 which indicated that POD may alleviate the process of fibrosis by the Fyn/Stat3 signaling pathway. Furthermore, the gain of function assay by lentivirus-mediated exogenous forced expression of Fyn abrogated the therapeutic effect of the POD on renal fibrosis and inflammation. Collectively, it can be concluded that POD exerted a protective effect on renal fibrosis by mediating Fyn/Stat3 signaling pathway.
Assuntos
Nefropatias , Obstrução Ureteral , Camundongos , Fibrose , Inflamação/metabolismo , Rim/metabolismo , Rim/patologia , Nefropatias/tratamento farmacológico , Nefropatias/patologia , Proteínas Proto-Oncogênicas c-fyn/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fyn/metabolismo , Células RAW 264.7/efeitos dos fármacos , Células RAW 264.7/metabolismo , Transdução de Sinais , Fator de Transcrição STAT3/metabolismo , Obstrução Ureteral/complicações , Obstrução Ureteral/tratamento farmacológico , Obstrução Ureteral/metabolismo , AnimaisRESUMO
As a traditional Chinese medicine, Rhododendron molle G. Don has a long history of treating rheumatoid arthritis. In this study, RAW 264.7 cells induced by lipopolysaccharide (LPS) were established as cell inflammatory model to evaluate the anti-inflammatory activity of chloroform extract from R. molle leaves (CERL), ethyl acetate extract from R. molle leaves (EERL) and butanol extract from R. molle leaves (BERL) and analyze the potential anti-inflammatory components of R. molle. Potential anti-inflammatory components analysis of CERL were performed by HPLC and UHPLC-Q-TOF-MS. Prediction of potential anti-inflammatory components by molecular docking experiments. Compared with negative control group, 25â µg/mL CERL could reduce the release level of NO by 62 %, and the mRNA expression levels of COX-2, IL-6, IL-1ß and TNF-α were reduced by 69.74 %, 86.25 %, 77.94 % and 56.80 %, respectively. Western-Blot showed similar results. CERL, EERL and BERL exerted their inhibitory activity in dose-dependent manner. All results showed that the higher the concentration, the better the anti-inflammatory activity. CERL showed the best inhibitory activity, the second was EERL, and then was BERL. 21 terpenoids and 4 flavonoids were identified in CERL by UHPLC-Q-TOF-MS. Molecular docking results showed that triterpenoids in CERL had better interaction with target proteins (TNF-α, IL-1ß). It indicated that triterpenoids may be potential anti-inflammatory components of R. molle leaves. This study explored the anti-inflammatory activities of CERL, EERL, BERL, which laid a foundation for further promoting the clinical application of R. molle.
Assuntos
Extratos Vegetais , Rhododendron , Animais , Camundongos , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Lipopolissacarídeos/farmacologia , Simulação de Acoplamento Molecular , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Células RAW 264.7/efeitos dos fármacos , Rhododendron/química , Fator de Necrose Tumoral alfaRESUMO
Abrus mollis Hance (Leguminosae) has a variety of biological activities, including anti-inflammatory, antioxidant, antibacterial, antiviral, and antitumor activities. However, the specific substances responsible for the anti-inflammatory effects are unknown. Abrusamide H (BJBS) is a truxillic acid derivative obtained from the leaves of Abrus mollis Hance and has potential anti-inflammatory effects. In this study, we aimed to estimate the potential effect and mechanism of BJBS in inflammation by establishing lipopolysaccharide (LPS)-stimulated RAW264.7 cells inâ vitro and an injured zebrafish tail fin inâ vivo. The RAW264.7 cells were treated with different concentrations of BJBS after LPS stimulation. The production of nitric oxide (NO) was detected by Griess reaction, and reactive oxygen species (ROS) were detected by an ROS assay kit. The levels of proinflammatory cytokines, including interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and interleukin 18 (IL-18) were measured by ELISA. Results showed that BJBS at all concentrations inhibited the proliferation of RAW264.7 macrophages after LPS stimulation by cell counting kit-8 and the production of NO and ROS. In the BJBS treatment group, the levels of IL-6, TNF-α, IL-1ß, and IL-18 decreased in a concentration-dependent manner. The results inâ vivo showed that no significant difference in the survival of zebrafish between the BJBS and blank groups and BJBS inhibited the migration and aggregation of zebrafish neutrophils in a dose-dependent manner in inflammation induced by tail transection-induced inflammation. In conclusion, BJBS inhibited the production of NO and ROS, decreased the levels of secreted IL-6, TNF-α, IL-1ß, and IL-18, and reduced the migration and aggregation of zebrafish neutrophils.
Assuntos
Anti-Inflamatórios , Peixe-Zebra , Animais , Camundongos , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Citocinas , Fabaceae/química , Inflamação/tratamento farmacológico , Inflamação/induzido quimicamente , Interleucina-18/efeitos adversos , Interleucina-6 , Lipopolissacarídeos/farmacologia , Óxido Nítrico , Células RAW 264.7/efeitos dos fármacos , Células RAW 264.7/metabolismo , Espécies Reativas de Oxigênio , Fator de Necrose Tumoral alfa , Peixe-Zebra/metabolismoRESUMO
Four new streptoglycerides E-H (1-4), with a rare 6/5/5/-membered ring system, were isolated from a marine-derived actinomycete Streptomyces specialis. The structures of 1-4 were elucidated by detailed analysis of HRESIMS, 1D and 2D NMR data and ECD spectra as well as comparison of their spectroscopic data with those reported in literature. Compounds 1-4 showed significant anti-inflammatory activity by inhibiting lipopolysaccharide (LPS)-induced nitric oxide (NO) production in Raw 264.7 cells with IC50 values ranging from 3.5 to 10.9 µM. Especially, 2 suppressed mRNA expression levels of iNOS and IL-6 without cytotoxicity.
Assuntos
Anti-Inflamatórios/farmacologia , Policetídeos/farmacologia , Streptomyces , Animais , Anti-Inflamatórios/química , Organismos Aquáticos , Sedimentos Geológicos , Concentração Inibidora 50 , Interleucina-6/metabolismo , Camundongos , Óxido Nítrico Sintase Tipo II/metabolismo , Policetídeos/química , Células RAW 264.7/efeitos dos fármacosRESUMO
In this study, Chinese yam polysaccharide (CYP) was isolated from yam by hydroextraction and alcoholic precipitation. Subsequently, the chlorosulfate-pyridine (CSA-Pyr) method was used to obtain the sulfated Chinese yam polysaccharide derivative (S-CYP) to evaluate its immunomodulatory activity in RAW 264.7 cells and to investigate its mechanism of action. The results revealed that the sulfated modification altered the physicochemical properties of CYP but had no impact on the main chain structure. S-CYP demonstrated excellent immunomodulatory activity by increasing the viability of RAW 264.7 macrophage cells and stimulating the production of reactive oxygen species (ROS), nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin (IL)-6. Moreover, signal transduction experiments showed that S-CYP induced the activation of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways through toll-like receptor 4 (TLR4), dramatically increasing p-ERK, p-JNK and p-p38 proteins. Meanwhile, immunofluorescence results showed that S-CYP could significantly promote the entry of NF-κB p65 into the nucleus, which is essential for triggering the NF-κB pathway. Furthermore, blocking antibody experiments revealed that specific inhibitors of TLR4, MAPKs, and NF-κB suppressed the generation of TNF-α and IL-6 in RAW 264.7 cells. These findings suggested that both CYP and S-CYP could be used as immunomodulatory agents and may have potential application prospects in the food and pharmaceutical industries.
Assuntos
Dioscorea , Medicamentos de Ervas Chinesas/farmacologia , Fatores Imunológicos/farmacologia , Polissacarídeos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Medicamentos de Ervas Chinesas/química , Alimento Funcional , Fatores Imunológicos/química , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Polissacarídeos/química , Células RAW 264.7/efeitos dos fármacos , Receptor 4 Toll-Like/metabolismoRESUMO
Microbe-based cancer immunotherapy has recently emerged as a hot topic for cancer treatment. However, serious limitations remain including infection associated side-effect and unsatisfactory outcomes in clinic trials. Here, we fabricate different sizes of nano-formulations derived from yeast cell wall (YCW NPs) by differential centrifugation. The induction of anticancer immunity of our formulations appears to inversely correlate with their size due to the ability to accumulate in tumor-draining lymph node (TDLN). Moreover, we use a percolation model to explain their distribution behavior toward TDLN. The abundance and functional orientation of each effector component are significantly improved not only in the microenvironment in tumor but also in the TDLN following small size YCW NPs treatment. In combination with programmed death-ligand 1 (PD-L1) blockade, we demonstrate anticancer efficiency in melanoma-challenged mice. We delineate potential strategy to target immunosuppressive microenvironment by microbe-based nanoparticles and highlight the role of size effect in microbe-based immune therapeutics.
Assuntos
Imunoterapia/métodos , Linfonodos/efeitos dos fármacos , Melanoma Experimental/terapia , Nanopartículas/administração & dosagem , Saccharomyces cerevisiae/química , Neoplasias Cutâneas/terapia , Aloenxertos , Animais , Antígeno B7-H1/antagonistas & inibidores , Antígeno B7-H1/genética , Antígeno B7-H1/imunologia , Linhagem Celular Tumoral , Parede Celular/química , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Feminino , Regulação Neoplásica da Expressão Gênica , Injeções Intralesionais , Linfonodos/imunologia , Linfonodos/patologia , Ativação de Macrófagos/efeitos dos fármacos , Melanoma Experimental/genética , Melanoma Experimental/mortalidade , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos C57BL , Nanopartículas/química , Tamanho da Partícula , Células RAW 264.7/efeitos dos fármacos , Células RAW 264.7/imunologia , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/mortalidade , Neoplasias Cutâneas/patologia , Análise de Sobrevida , Quinase Syk/antagonistas & inibidores , Quinase Syk/genética , Quinase Syk/imunologia , Receptor 2 Toll-Like/antagonistas & inibidores , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/imunologia , Carga Tumoral/efeitos dos fármacos , Microambiente Tumoral/efeitos dos fármacosRESUMO
Atlantic cod (Gadus morhua) is one of the most important fishes in the world with high nutritional value and economic value. However, the impact and underlying mechanism of the G. morhua peptides (GMPs) on osteoclastogenesis and bone mineral density (BMD) regulation remain unclear. The purpose of this study was to investigate the effects of GMPs on osteoclast formation and anti-osteoporosis activity in vitro and in vivo. The results showed that GMPs significantly reduced receptor activator of nuclear factor (RANKL) induced tartrate-resistant acid phosphatase (TRAP) activity, and decreased the expression of osteoclast regulatory factors c-Fos and NFATc1 by inhibiting the activation of MAPK and NF-κB pathways, and thereby inhibiting osteoclast formation and bone resorption. In vivo, GMP protects mice against ovariectomy-induced bone loss by regulating the balance of major factors released in bone formation and resorption. Taken together, GMP could be a potential candidate or dietary supplement for the prevention of osteoporosis.
Assuntos
Conservadores da Densidade Óssea/uso terapêutico , Alimento Funcional , Gadus morhua , Peptídeos/uso terapêutico , Animais , Densidade Óssea/efeitos dos fármacos , Conservadores da Densidade Óssea/farmacologia , Modelos Animais de Doenças , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Osteoclastos/efeitos dos fármacos , Osteoporose/induzido quimicamente , Osteoporose/prevenção & controle , Ovariectomia , Peptídeos/farmacologia , Ligante RANK , Células RAW 264.7/efeitos dos fármacosRESUMO
Inflammation is a double-edged sword, as it can have both protective effects and harmful consequences, which, combined with oxidative stress (OS), can lead to the development of deathly chronic inflammatory conditions. Over the years, research has evidenced the potential of marine sponges as a source of effective anti-inflammatory therapeutic agents. Within this framework, the purpose of this study was to evaluate the antioxidant and the anti-inflammatory potential of the marine sponge Cliona celata. For this purpose, their organic extracts (C1-C5) and fractions were evaluated concerning their radical scavenging activity through 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and anti-inflammatory activity through a (lipopolysaccharides (LPS)-induced inflammation on RAW 264.7 cells) model. Compounds present in the two most active fractions (F5 and F13) of C4 were tentatively identified by gas chromatography coupled to mass spectrometry (GC-MS). Even though samples displayed low antioxidant activity, they presented a high anti-inflammatory capacity in the studied cellular inflammatory model when compared to the anti-inflammatory standard, dexamethasone. GC-MS analysis led to the identification of n-hexadecanoic acid, cis-9-hexadecenal, and 13-octadecenal in fraction F5, while two major compounds, octadecanoic acid and cholesterol, were identified in fraction F13. The developed studies demonstrated the high anti-inflammatory activity of the marine sponge C. celata extracts and fractions, highlighting its potential for further therapeutic applications.
Assuntos
Antineoplásicos/farmacologia , Poríferos , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Antineoplásicos/química , Antioxidantes/química , Antioxidantes/farmacologia , Organismos Aquáticos , Linhagem Celular Tumoral/efeitos dos fármacos , Células HT29/efeitos dos fármacos , Humanos , Lipopolissacarídeos , Camundongos , Portugal , Células RAW 264.7/efeitos dos fármacosRESUMO
Plant-derived antimalarials are indispensable for malaria treatment and a platform for new drugs. The present study explores sinigrin, for malaria using in vitro, in silico and in vivo strategies and the immune response generated after administration. The compound exhibited promising activity against chloroquine (CQ)-resistant (RKL-9) IC50 5.14 µg/mL and CQ-sensitive (3D7) IC50 5.47 µg/mL strains of P. falciparum and was safe in both in vitro (CC50 > 640 µg/mL) and in vivo (LD50 > 2 g/kg) toxicity studies. In addition, virtual screening showed hydrogen bonding, hydrophobic and van der Waals interactions with amino acid residues of 3BPM (falcipain-3). In vivo studies revealed promising antimalarial activity of sinigrin (200 mg/kg) with 87.44% chemo-suppression on day 5 and significantly (p < 0.0001) enhanced the mean survival time (21 ± 4.74 days) in contrast to the infected control (5.4 ± 1.14 days). In combination therapy, sinigrin (100 mg/kg and 200 mg/kg) augmented the efficacy of artesunate (AS 50 mg/kg) with 100% survival and no recrudescence. These observations are further corresponded and supported by DLC, NO production, cytokine analysis, biochemical and histopathological studies. Treatment with the combination resulted in a regulated interplay of immune cells and cytokines aiding in parasite clearance in addition to its specific inhibitory activity. We report the antimalarial activity of sinigrin first time with best D-score against falcipain-3. These findings highlight sinigrin as a HIT molecule, which may potentially be used in drug and vaccine development approaches.
Assuntos
Antimaláricos/uso terapêutico , Artesunato/uso terapêutico , Cisteína Endopeptidases/metabolismo , Glucosinolatos/uso terapêutico , Malária/tratamento farmacológico , Animais , Antimaláricos/administração & dosagem , Artesunato/administração & dosagem , Cisteína Endopeptidases/efeitos dos fármacos , Citocinas/metabolismo , Quimioterapia Combinada , Feminino , Glucosinolatos/administração & dosagem , Contagem de Leucócitos , Malária/imunologia , Malária Falciparum/tratamento farmacológico , Malária Falciparum/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Simulação de Acoplamento Molecular , Óxido Nítrico/metabolismo , Plasmodium berghei/efeitos dos fármacos , Plasmodium falciparum/efeitos dos fármacos , Células RAW 264.7/efeitos dos fármacos , Células RAW 264.7/metabolismoRESUMO
BACKGROUND: The present study extensively aimed to evaluate the underlying mechanism of the immunomodulatory and anti-inflammatory effects of Phellinus linteus mycelium (PLM). METHODS: To assess whether PLM influences the production of markers related to inflammation, Lipopolysaccharide (LPS)-stimulated RAW264.7 cells were treated with PLM (50, 100, 200, and 500 µg/mL). Splenocyte, thymus, peritoneal exudate cells (PEC), and peripheral blood mononuclear cells (PBMC) were isolated from the Balb/c mice treated with Korean red ginseng or PLM once a day for 5 weeks. Moreover, all mice except normal mice were stimulated with 10% proteose peptone (PP) treated 3 days before the sacrifice and 2% starch treated 2 days before the sacrifice. Subsequently, the cytotropic substance was evaluated by using flow cytometry analysis and ELISA assay. RESULTS: PLM200 treatment significantly suppressed the production of nitric oxide (NO) and prostaglandin E2 (PGE2) and inhibited the release of proinflammatory cytokines such as interleukin (IL)-6, IL-1ß, and tumor necrosis factor (TNF)-α dose-dependently in the LPS-stimulated RAW264.7 cells. PLM200 supplementation showed a significant increase in IL-2, IL-12, and interferon (IFN)-γ production and upregulated the ratio of IFN-γ (T-helper type 1, Th1) to IL-4 (T-helper type 2, Th2) in splenocytes. After PLM200 treatment, the significant elevation of CD4+CD25+, CD4+&CD8+, and CD4+CD69+ treatment were detected in thymus. Moreover, CD4+ and CD4+CD69+ in PBMC and CD69+ in PEC were also shown in a significant increase. CONCLUSIONS: Taken together, these results showed an immunomodulatory effect of PLM about an elevated INF-γ/IL4 ratio, as an index of Th1/Th2, as well as the anti-inflammatory effect in the LPS-stimulated RAW264.7 cells. Therefore, our findings demonstrate that PLM possesses immunostimulatory and anti-inflammatory effects.
Assuntos
Anti-Inflamatórios/farmacologia , Agentes de Imunomodulação/farmacologia , Extratos Vegetais/farmacologia , Animais , Austrália , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Phellinus , Células RAW 264.7/efeitos dos fármacos , República da CoreiaRESUMO
Cigarette smoke (CS) exposure is a risk factor for chronic obstructive pulmonary disease (COPD). CS exposure impairs the ability of killing pathogens in macrophages, which might be due to the abnormal apoptosis induced by CS. This study explored the effects and mechanisms of cigarette smoke extract (CSE) on the apoptosis of macrophages in vitro. Raw264.7 cells were treated with CSE at different concentrations, and viability and apoptosis of cells was accessed. The protein expression was detected by western blot. The intracellular Ca2+ level was evaluated by Fluo-4 AM probe assay. CSE induced the apoptosis and increased the expression of cleaved caspase 3, which were attenuated by a caspase inhibitor. CSE increased the expression of CHOP, BiP and P-eif2α, and the inhibitor of endoplasmic reticulum stress (ERS) decreased the apoptosis induced by CSE. Phosphorylation levels of P38, JNK and ERK1/2 were increased following incubation with CSE. Only P38 inhibitor significantly reduced apoptosis induced by CSE, while ERK1/2 inhibitor promoted apoptosis. Phosphorylation of STAT1 at Ser727 was activated by CSE and attenuated by the P38 inhibitor. Finally, CSE increased the level of intracellular Ca2+, and calcium chelator partly attenuated the apoptosis and phosphorylation of P38 and STAT1 induced by CSE. CSE induced a caspase 3-dependent apoptosis in Raw264.7 cells via ERS and intracellular Ca2+/P38/STAT1 pathway.
Assuntos
Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Fumar Cigarros/efeitos adversos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Fator de Transcrição STAT1/metabolismo , Fumaça/efeitos adversos , Animais , Western Blotting , Camundongos , Células RAW 264.7/efeitos dos fármacos , Produtos do Tabaco/efeitos adversosRESUMO
Cancer-related bone erosion occurs frequently in bone metastasis and is associated with severe complications such as chronic bone pain, fractures, and lower survival rates. In recognition of the fact that the darkness hormone melatonin is capable of regulating bone homeostasis, we explored its therapeutic potential in bone metastasis. We found that melatonin directly reduces osteoclast differentiation, bone resorption activity and promotes apoptosis of mature osteoclasts. We also observed that melatonin inhibits RANKL production in lung and prostate cancer cells by downregulating the p38 MAPK pathway, which in turn prevents cancer-associated osteoclast differentiation. In lung and prostate bone metastasis models, twice-weekly melatonin treatment markedly reduced tumor volumes and numbers of osteolytic lesions. Melatonin also substantially lowered the numbers of TRAP-positive osteoclasts in tibia bone marrow and RANKL expression in tumor tissue. These findings show promise for melatonin in the treatment of bone metastases.
Assuntos
Neoplasias Ósseas/tratamento farmacológico , Melatonina/farmacologia , Osteoclastos/efeitos dos fármacos , Ligante RANK/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Animais , Células da Medula Óssea/efeitos dos fármacos , Neoplasias Ósseas/genética , Neoplasias Ósseas/patologia , Neoplasias Ósseas/secundário , Reabsorção Óssea/tratamento farmacológico , Reabsorção Óssea/genética , Reabsorção Óssea/patologia , Diferenciação Celular/efeitos dos fármacos , Modelos Animais de Doenças , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Osteoclastos/patologia , Osteólise/tratamento farmacológico , Osteólise/patologia , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Células RAW 264.7/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Fosfatase Ácida Resistente a Tartarato/genéticaRESUMO
Microalgae are known as a producer of proteins and lipids, but also of valuable compounds for human health benefits (e.g., polyunsaturated fatty acids (PUFAs); minerals, vitamins, or other compounds). The overall objective of this research was to prospect novel products, such as nutraceuticals from microalgae, for application in human health, particularly for metabolic diseases. Chlorella vulgaris and Chlorococcum amblystomatis were grown autotrophically, and C. vulgaris was additionally grown heterotrophically. Microalgae biomass was extracted using organic solvents (dichloromethane, ethanol, ethanol with ultrasound-assisted extraction). Those extracts were evaluated for their bioactivities, toxicity, and metabolite profile. Some of the extracts reduced the neutral lipid content using the zebrafish larvae fat metabolism assay, reduced lipid accumulation in fatty-acid-overloaded HepG2 liver cells, or decreased the LPS-induced inflammation reaction in RAW264.7 macrophages. Toxicity was not observed in the MTT assay in vitro or by the appearance of lethality or malformations in zebrafish larvae in vivo. Differences in metabolite profiles of microalgae extracts obtained by UPLC-LC-MS/MS and GNPS analyses revealed unique compounds in the active extracts, whose majority did not have a match in mass spectrometry databases and could be potentially novel compounds. In conclusion, microalgae extracts demonstrated anti-obesity, anti-steatosis, and anti-inflammatory activities and could be valuable resources for developing future nutraceuticals. In particular, the ultrasound-assisted ethanolic extract of the heterotrophic C. vulgaris significantly enhanced the anti-obesity activity and demonstrated that the alteration of culture conditions is a valuable approach to increase the production of high-value compounds.
Assuntos
Anti-Inflamatórios/farmacologia , Fármacos Antiobesidade/farmacologia , Chlorella vulgaris , Microalgas , Animais , Anti-Inflamatórios/química , Fármacos Antiobesidade/química , Organismos Aquáticos , Células Hep G2/efeitos dos fármacos , Humanos , Larva/efeitos dos fármacos , Camundongos , Células RAW 264.7/efeitos dos fármacos , Peixe-ZebraRESUMO
Sargassum horneri is an invasive brown seaweed that grows along the shallow coastal areas of the Korean peninsula, which are potentially harmful to fisheries and natural habitats in the areas where it is accumulated. Therefore, the author attempted to evaluate the anti-inflammatory mechanism of Sargachromenol isolated from S. horneri against particulate matter (PM)-stimulated RAW 264.7 macrophages. PM is a potent inducer of respiratory diseases such as lung dysfunctions and cancers. In the present study, the anti-inflammatory properties of Sargachromenol were validated using enzyme-linked immunosorbent assay (ELISA), Western blots, and RT-qPCR experiments. According to the results, Sargachromenol significantly downregulated the PM-induced proinflammatory cytokines, Prostaglandin E2 (PGE2), and Nitric Oxide (NO) secretion via blocking downstream activation of Toll-like receptor (TLR)-mediated nuclear factor kappa B (NF-κB) and MAPKs phosphorylation. Thus, Sargachromenol is a potential candidate for innovation in various fields including pharmaceuticals, cosmeceuticals, and functional food.
Assuntos
Anti-Inflamatórios/farmacologia , Benzopiranos/farmacologia , Extratos Vegetais/farmacologia , Sargassum , Animais , Anti-Inflamatórios/química , Organismos Aquáticos , Benzopiranos/química , Humanos , Macrófagos/metabolismo , Camundongos , Material Particulado , Extratos Vegetais/química , Células RAW 264.7/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Receptores Toll-Like/metabolismoRESUMO
A promising use of bismuth nanoparticles (BiNPs) for different biomedical applications leads to a search for the elucidation of their toxicity mechanisms, since toxicity studies are still at early stage. In the current study, cytotoxic effects of BiNPs produced by laser ablation in solution (LASiS) was investigated in the murine macrophage line RAW 264.7. The cells were exposed to 0.01-50 µg ml-1 of BiNPs for 24 and 48 h and then cytotoxicity assays were performed. Decrease of MTT conversion to formazan and of cell attachment were observed with no effects on cell proliferation. No loss of membrane integrity or significant changes of ROS and RNS levels were observed in exposed cells. Foremost, increased phagocytic activity and DNA repair foci occurred for cells exposed to BiNPs. These effects are important findings that must be considered in the case of biomedical application of BiNPs, since inappropriate macrophages activation and inactivation may lead to immunotoxicity. Bismuth nanoparticles (BiNPs) produced by laser ablation in solution and stabilized with BSA decrease enzyme-dependent MTT conversion to formazan and increase phagocytic activity and DNA repair foci in murine macrophage line RAW 264.7 when exposed to 50 µg ml-1. These effects are findings that should be considered in the case of biomedical application of BiNPs, since inappropriate macrophages activation and inactivation may lead to immunotoxicity.
Assuntos
Bismuto/toxicidade , Formazans/química , Macrófagos/efeitos dos fármacos , Nanopartículas Metálicas/química , Células RAW 264.7/efeitos dos fármacos , Animais , Bismuto/química , Adesão Celular , Ciclo Celular , Proliferação de Células , Sobrevivência Celular , DNA/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Reparo do DNA , Lasers , Macrófagos/citologia , Camundongos , Fagocitose , Células RAW 264.7/citologia , Espécies Reativas de Oxigênio , Sais de Tetrazólio/química , Tiazóis/químicaRESUMO
Apigenin (4',5,7-trihydroxyflavone, flavonoid) is a phenolic compound that is known to reduce the risk of chronic disease owing to its low toxicity. The first study on apigenin analyzed its effect on histamine release in the 1950s. Since then, anti-mutation and antitumor properties of apigenin have been widely reported. In the present study, we evaluated the apigenin-mediated amelioration of skin disease and investigated its applicability as a functional ingredient, especially in cosmetics. The effect of apigenin on RAW264.7 (murine macrophage), RBL-2H3 (rat basophilic leukemia), and HaCaT (human immortalized keratinocyte) cells were analyzed. Apigenin (100 µM) significantly inhibited nitric oxide (NO) production, cytokine expression (interleukin (IL)-1ß, IL6, cyclooxygenase (COX)-2, and inducible nitric oxide synthase [iNOS]), and phosphorylation of mitogen-activated protein kinase (MAPK) signal molecules, including extracellular signal-regulated kinase (ERK) and c-Jun N-terminal protein kinase (JNK) in RAW264.7 cells. Apigenin (30 M) also inhibited the phosphorylation of signaling molecules (Lyn, Syk, phospholipase Cγ1, ERK, and JNK) and the expression of high-affinity IgE receptor FcεRIα and cytokines (tumor necrosis factor (TNF)-α, IL-4, IL-5, IL-6, IL-13, and COX-2) that are known to induce inflammation and allergic responses in RBL-2H3 cells. Further, apigenin (20 µM) significantly induced the expression of filaggrin, loricrin, aquaporin-3, hyaluronic acid, hyaluronic acid synthase (HAS)-1, HAS-2, and HAS-3 in HaCaT cells that are the main components of the physical barrier of the skin. Moreover, it promoted the expression of human ß-defensin (HBD)-1, HBD-2, HBD-3, and cathelicidin (LL-37) in HaCaT cells. These antimicrobial peptides are known to play an important role in the skin as chemical barriers. Apigenin significantly suppressed the inflammatory and allergic responses of RAW264.7 and RBL cells, respectively, and would, therefore, serve as a potential prophylactic and therapeutic agent for immune-related diseases. Apigenin could also be used to improve the functions of the physical and chemical skin barriers and to alleviate psoriasis, acne, and atopic dermatitis.
Assuntos
Apigenina/farmacologia , Dermatopatias/tratamento farmacológico , Animais , Antialérgicos/metabolismo , Antialérgicos/farmacologia , Anti-Inflamatórios/metabolismo , Anti-Inflamatórios/farmacologia , Apigenina/metabolismo , Linhagem Celular , Citocinas/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Proteínas Filagrinas , Células HaCaT/efeitos dos fármacos , Humanos , Imunoglobulina E/metabolismo , Interleucina-1beta/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Mastócitos/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fosforilação/efeitos dos fármacos , Células RAW 264.7/efeitos dos fármacos , Ratos , Receptores de IgE/genética , Transdução de Sinais/efeitos dos fármacos , Pele/efeitos dos fármacos , Pele/metabolismo , Fenômenos Fisiológicos da Pele/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Balanophora polyandra Griff. (B. polyandra) is a folk medicine used as an antipyretic, antidote, haemostatic, dressing and haematic tonic, for the treatment of gonorrhea, syphilis, wounds, and the bleeding of the alimentary tract by the local people in China. This study was designed to investigate the effects of B. polyandra on dextran sulfate sodium (DSS)-treated colitis mice in vivo and lipopolysaccharide (LPS)-induced RAW 264.7 macrophages in vitro. Mice were induced with B. polyandra total extract (BPE, 250 and 1000 mg kg-1) and B. polyandra polysaccharides (BPP, 100 and 400 mg kg-1) for 22 days and treated with 3.5% DSS in their drinking water for the last 7 days and the LPS-induced RAW264.7 macrophages were treated with BPE (100 µg ml-1) and BPP (100 µg ml-1). Mice treated with DSS developed severe mucosal colitis, with a marked distortion and crypt loss of colonic surface epithelium and a colonic shortening. B. polyandra significantly inhibited colonic shortening and reduced the severity of colitis in the colon and lowered the colonic inflammation score (p < 0.05) and decreased the expression of interleukin (IL)-1ß, tumor necrosis factor (TNF-α), inducible nitric oxide synthase (iNOS), and anti-serum amyloid A3 (SAA3) as well as the pro-inflammatory chemokine C-X-C motif chemokine 10 (CXCL10). B. polyandra also significantly suppressed the activation of nucleotide-binding domain like receptor protein 3 (NLRP3) inflammasome and the nuclear factor kB (NF-κB). These results suggest that dietary intake of B. polyandra ameliorates colitis. Such activities of B. polyandra in humans remain to be investigated.
Assuntos
Balanophoraceae , Colite/prevenção & controle , Extratos Vegetais/uso terapêutico , Animais , Colite/induzido quimicamente , Colite/metabolismo , Sulfato de Dextrana , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fitoterapia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Células RAW 264.7/efeitos dos fármacosRESUMO
Acai fruit is recognized for its health promoting properties. However, there is still a need to address the effects of industrial processing on this fruit. In this study, phenolic content, anti-inflammatory properties and dermal wound repair properties of 20 acai samples, before and after industrial processing, from various Amazon regions were investigated. Acai pulp was rich in total phenolics (18.9-58.8 mg g-1) and proanthocyanins (9.8-43.1 mg g-1), but contained trace anthocyanins (up to 0.1 mg g-1). Industrially processed samples lost substantial amounts of proanthocyanidins (up to 83.2%), while the anthocyanins inherently present were greatly enriched after processing (20-fold higher). Non-processed acai pulp extracts protected against early inflammation response which was correlated with proanthocyanidins, by significantly inhibiting nitric oxide production and suppressing pro-inflammatory gene expression including interleukin-1ß, cyclooxygenase-2, nitric oxide synthase, and interleukin-6. The promotion of dermal wound repair of acai seed and pulp extracts was mainly contributed by anthocyanins and other bioactive compounds. The anti-inflammatory effect was diminished but wound healing effect was retained after pulp processing, suggesting the processing technology needs to be improved to maintain biological properties of acai fruit.
Assuntos
Anti-Inflamatórios/farmacologia , Arecaceae , Extratos Vegetais/farmacologia , Cicatrização/efeitos dos fármacos , Animais , Brasil , Indústria Alimentícia , Frutas , Humanos , Camundongos , Fitoterapia , Extratos Vegetais/química , Polifenóis/química , Células RAW 264.7/efeitos dos fármacosRESUMO
Akebia trifoliata fruits, a kind of popular edible berry in Asia, are widely consumed as daily fruits or functional foods. Our previous study found several bioactives from Akebia trifoliata pericarp extract (APE), and preliminarily investigated their anti-inflammatory activity. However, the underlying mechanism of APE for the observed anti-inflammatory effects is still unknown. Thus, the bioactive profiles and anti-inflammatory mechanism of APE were investigated by a combination of chemical assays: UPLC-LTQ-Orbitrap/MS technique, lipopolysaccharide (LPS)-induced RAW264.7 cells and DSS-induced mouse model. The results indicated that the phenolic acids and terpenoids are major bioactives of APE, which could inhibit the production of nitric oxide (NO) and prostaglandin E2 (PGE2) by blocking the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in RAW264.7 cells as well as reduce the expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß), and suppress the phosphorylation of p-65, IκBα and mitogen-activated protein kinase (MAPKs including p38, ERKs, JNKs) proteins both in vitro and in vivo. Furthermore, APE treatment could regulate gut microbiota by increasing the richness of Rikenellaceae and Lactobacillaceae and reducing that of Lachnospiraceae and Ruminococcaceae. In summary, these findings clearly demonstrated that APE mitigates inflammation by restraining the production of cytokines through nuclear factor-κB (NF-κB) and MAPK signaling pathways, and altering gut microbiota, and therefore, this could be a potential functional food for the treatment and prevention of inflammatory bowel diseases.
Assuntos
Anti-Inflamatórios/uso terapêutico , Frutas , Doenças Inflamatórias Intestinais/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacologia , Modelos Animais de Doenças , Microbioma Gastrointestinal/efeitos dos fármacos , Doenças Inflamatórias Intestinais/induzido quimicamente , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , Fitoterapia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Células RAW 264.7/efeitos dos fármacosRESUMO
Probiotics are known to modulate gut microbiota, intestinal barrier function and host immune response, but due to the species and strain specific response their mechanisms are not clearly understood. Thus, the present study was designed to isolate, assess the anti-inflammatory potential and underlying modulatory mechanisms of indigenous probiotics in murine macrophage cell line, RAW 264.7. Forty lactic acid bacteria (LAB) were isolated from different sources and monitored for their anti-inflammatory potential against lipopolysaccharide (LPS) induced inflammatory stress employing RAW 264.7 cells. Among these isolates, only four LAB isolates exhibited more than 90% nitric oxide inhibition and possessed the probiotic attributes. Further, these selected LAB isolates reduced the level of pro-inflammatory cytokines, TNF-α, IL-1ß and IL-6, inhibited the phosphorylation of Mitogen Activated Protein Kinases (MAPKs) i.e. p38 MAPK, ERK1/2 and SAPK/JNK and expression of cyclooxygenase-2 (COX-2) in LPS stimulated RAW 264.7 cells. The in vitro analysis suggested that the selected probiotic isolates attenuated the LPS-induced inflammation by downregulating MAPK pathway vis-a-vis inhibiting COX-2 and can be employed as anti-inflammatory agents in various inflammatory diseases.