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










Base de dados
Intervalo de ano de publicação
1.
Int J Mol Sci ; 22(16)2021 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-34445462

RESUMO

Liver cancer is the sixth most common cancer worldwide with high morbidity and mortality. Programmed death ligand 1 (PD-L1) is a major ligand of programmed death 1 receptor (PD1), and PD1/PD-L1 checkpoint acts as a negative regulator of the immune system. Cancers evade the host's immune defense via PD-L1 expression. This study aimed to investigate the effects of tumor-related cytokines, interferon gamma (IFNγ), and tumor necrosis factor alpha (TNFα) on PD-L1 expression in human hepatocellular carcinoma cells, HepG2. Furthermore, as atorvastatin, a cholesterol-lowering agent, is documented for its immunomodulatory properties, its effect on PD-L1 expression was investigated. In this study, through real-time RT-PCR, Western blot, and immunocytochemistry methods, PD-L1 expression in both mRNA and protein levels was found to be synergistically upregulated in HepG2 by a combination of IFNγ and TNFα, and STAT1 activation was mainly responsible for that synergistic effect. Next, atorvastatin can inhibit the induction of PD-L1 by either IFNγ alone or IFNγ/TNFα combination treatment in HepG2 cells. In conclusion, in HepG2 cells, expression of PD-L1 was augmented by cytokines in the tumor microenvironment, and the effect of atorvastatin on tumor immune response through inhibition of PD-L1 induction should be taken into consideration in cancer patients who have been prescribed atorvastatin.


Assuntos
Atorvastatina/farmacologia , Antígeno B7-H1/imunologia , Carcinoma Hepatocelular/imunologia , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/imunologia , Proteínas de Neoplasias/imunologia , Antígeno B7-H1/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/imunologia , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Proteínas de Neoplasias/genética
2.
FASEB J ; 35(5): e21487, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33811705

RESUMO

Chondrosarcoma is a cartilage-forming bone tumor, well known for intrinsic resistance to chemotherapy and radiotherapy. We have designed a targeted chondrosarcoma gene therapy using a bacteriophage (phage) particle to deliver therapeutic genes. Phage has no tropism for mammalian cells, allowing engineered phage to be targeted to specific cell surface receptors in cancer. We modified the phage capsid to display the RGD4C ligand on the pIII minor coat proteins to specifically bind to αvß3 or αvß5 integrin receptors. The endosomal escape peptide, H5WYG, was also displayed on recombinant pVIII major coat proteins to enhance gene delivery. Finally, a human tumor necrosis factor alpha (TNFα) therapeutic transgene expression cassette was incorporated into the phage genome. First, we found that human chondrosarcoma cells (SW1353) have high expression of αvß3, αvß5 integrin receptors, and both TNFα receptors. Targeted particle encoding a luciferase reporter gene efficiently and selectively mediated gene delivery to these cells. When SW1353 cells were treated with the targeted particle encoding a TNFα transgene, significant cell killing was evident and was associated with high expression of TNFα and apoptosis-related genes. In vivo, mice with established human chondrosarcoma showed suppression of tumors upon repetitive intravenous administrations of the targeted phage. These data show that our phage-based particle is a promising, selective, and efficient tool for targeted chondrosarcoma therapy.


Assuntos
Bacteriófagos/genética , Neoplasias Ósseas/terapia , Condrossarcoma/terapia , Técnicas de Transferência de Genes , Terapia Genética , Terapia por Fagos/métodos , Fator de Necrose Tumoral alfa/genética , Adulto , Animais , Apoptose , Neoplasias Ósseas/genética , Neoplasias Ósseas/patologia , Proliferação de Células , Condrossarcoma/genética , Condrossarcoma/patologia , Vetores Genéticos/administração & dosagem , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
3.
Brain Res Bull ; 172: 190-202, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33894297

RESUMO

Neuroinflammation-mediated microglial reactivity is a major process, which explains the increased risk of Alzheimer's disease (AD) development in patients with Type 2 diabetes mellitus (T2DM). Advanced glycation end products (AGEs), formed by hyperglycemic condition in diabetes, is characterized as an intermediary of brain injury with diabetes through induction of microglial reactivity. Here, we explored the effect of AGEs on microglial reactivity using BV2 as a model. The NF-κB, p38 and JNK pathways were found to be important mechanism in AGEs-induced BV2 microglial reactivity. NF-κB inhibitor (BAY-11-7082), p38 inhibitor (SB203580) and JNK inhibitor (SP600125) exhibited the potential inhibition of AGEs-induced NO production. We also found that the sesamin, a major lignan found in sesame seed oils, exerts an anti-inflammatory effect under AGEs-induced microglial reactivity via suppressing the phosphorylation of NF-κB, p38 and JNK pathways. Moreover, sesamin also ameliorated AGEs-induced-receptor for advanced glycation end products (RAGE) expression. Taken together, sesamin may be a promising phytochemical compound to delay inflammatory progress by AGEs microglia function. Similarly, inhibition of AGEs-induced microglial reactivity might be potential therapeutic targets of neuroinflammation-based mechanisms in T2DM link progressive AD.

4.
Int Immunopharmacol ; 86: 106759, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32663768

RESUMO

Programmed death ligand 1 (PD-L1) is overexpressed in some metastatic breast cancer subtypes, specifically triple-negative breast cancer (TNBC). This feature can assist in the eradication of anti-tumor immunity, thereby enhancing the survival of the tumor. This study aims to explore how sesamin affects PD-L1 expression in breast cancer cells and its related molecular mechanisms. We found high levels of expression of PD-L1 in both mRNA and protein levels in the TNBC cell line, MDA-MB231, but not in the luminal type-breast cancer cell line, MCF-7. We then demonstrated the tumor suppressive effect of sesamin, which induced the inhibition of cell proliferation in MDA-MB231 cells. Additionally, sesamin triggered PD-L1 downregulation (both mRNA and protein) through the inhibition of AKT, NF-κB and JAK/Stat signaling in MDA-MB231 cells. Moreover, the migration ability of MDA-MB231 cells was effectively diminished by sesamin via inhibition of the activation of MMP-9 and MMP-2. In summary, this study demonstrated that sesamin suppresses MDA-MB231 breast cancer cells' proliferation and migration; and decreases the expression of PD-L1 via the downregulation of AKT, NF-κB, and JAK/Stat signaling. Therefore, sesamin may be an effective alternative and novel therapeutic option for immunotherapy in breast cancer cells with high PD-L1 expression.


Assuntos
Adenocarcinoma/tratamento farmacológico , Antígeno B7-H1/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/tratamento farmacológico , Dioxóis/farmacologia , Lignanas/farmacologia , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Antígeno B7-H1/genética , Biomarcadores Tumorais/genética , Proliferação de Células , Feminino , Humanos , Janus Quinases/metabolismo , Células MCF-7 , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , NF-kappa B/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fatores de Transcrição STAT/metabolismo , Transdução de Sinais , Regulação para Cima
5.
Artigo em Inglês | MEDLINE | ID: mdl-32308720

RESUMO

The promotion of neurogenesis can be a promising strategy to improve and restore neuronal function in neurodegenerative diseases. Nerve growth factor (NGF) plays a key role in neurite outgrowth and synaptic formation during brain repair stage. Nowadays, there are several studies on the developing methods to enhance the endogenous NGF activity for treatment and restore the neuronal function. In this study, the potentiating effect of sesamin, a major lignan in sesame seeds (Sesamum indicum) and oil, on NGF-induced neurogenesis and its involved mechanisms were firstly reported. Sesamin effectively enhanced the PC12 neuron-like cell differentiation and neurite length under insufficient conditions of NGF. The neuronal markers including synaptophysin and growth-associated protein-43 along with the synaptic connections were significantly increased in combination treatment between sesamin and NGF. Moreover, sesamin also increased the level of phospho-ERK1/2 and SIRT1 protein, an important regulatory protein of the neurogenesis process. The neurogenesis was blocked by the specific SIRT1 inhibitor, JGB1741, suggesting that the neuritogenic effect of sesamin was associated with SIRT1 protein modulation. Taken together, the potentiating effect of sesamin on NGF-induced neurogenesis in this finding could be used for alternative treatment in neurodegenerative diseases, including Alzheimer's disease.

6.
J Neuroimmunol ; 341: 577164, 2020 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-32007785

RESUMO

SIRT1 exhibits inhibitory effects on microglial activation-induced neurodegeneration. Regulating SIRT1 may become a novel approach for curing neurodegenerative diseases. Protocatechuic acid (PA), a phenolic acid, has anti-neuroinflammatory effects. The effect of PA on SIRT1 in activated microglia remains unknown. Here, we examined whether PA has anti-inflammatory effects against microglial activation-induced neuronal cell death via regulating SIRT1 in microglia. We found that PA inhibited the release of inflammatory mediators in LPS-activated BV2 microglia via the SIRT1/NF-κB pathway and thereby attenuated microglial activation-induced PC12 cell apoptosis. This suggests that SIRT1 mediates the anti-neuroinflammatory effects of PA to ameliorate microglial activation-induced neuron death.


Assuntos
Anti-Inflamatórios/farmacologia , Hidroxibenzoatos/farmacologia , Microglia/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Animais , Apoptose/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Meios de Cultivo Condicionados/farmacologia , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Microglia/metabolismo , Microglia/ultraestrutura , NF-kappa B/fisiologia , Doenças Neurodegenerativas/tratamento farmacológico , Doenças Neurodegenerativas/imunologia , Doenças Neurodegenerativas/patologia , Óxido Nítrico/metabolismo , Células PC12 , Ratos , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/fisiologia
7.
Molecules ; 25(2)2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31936263

RESUMO

Programmed death ligand 1 (PD-L1) is overexpressed in the most aggressive breast cancer subtype, triple-negative breast cancer (TNBC), assisting the eradication of antitumor immunity, and thereby enhancing the survival of the tumor. This study explored how hesperidin affects PD-L1 expression, and thereby cancer progression in breast cancer cells. We found that MDA-MB231, the triple-negative breast adenocarcinoma cancer cell line, (high aggressiveness) has higher expression, in both mRNA and protein, of PD-L1 than that of the other breast cancer cell line, MCF-7 (low aggressiveness). Hesperidin inhibited cell proliferation in MDA-MB231 cells. Additionally, high expression of PD-L1 (both mRNA and protein) in aggressive cancer cells was strongly inhibited by hesperidin through inhibition of Akt and NF-κB signaling. Moreover, hesperidin treatment, by inhibiting activation of matrix metalloproteinases such as MMP-9 and MMP-2, suppressed the metastatic phenotype and cell migration in the PD-L1 high-expressing MDA-MB231 cells. In summary, hesperidin inhibits breast cancer cell growth through the inhibition of the expression of PD-L1 via downregulation of Akt and NF-κB signaling in TNBC. Moreover, hesperidin significantly suppresses cell migration of MDA-MB231 cells. Our findings reveal fresh insights into the anticancer effects of hesperidin which might have potential clinical implications.


Assuntos
Antígeno B7-H1/metabolismo , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Hesperidina/farmacologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Feminino , Hesperidina/química , Humanos , Metaloproteinases da Matriz/metabolismo , Modelos Biológicos , NF-kappa B/metabolismo , Invasividade Neoplásica , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo
8.
Neurotox Res ; 37(1): 111-125, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31485933

RESUMO

Neuroinflammation is a major factor in the pathogenesis of various neurodegenerative diseases. Microglia are resident macrophages that act as key mediators of inflammation in the brain. In response to inflammatory stimuli including lipopolysaccharide (LPS), microglial activation occurs immediately. Overproduction of inflammatory mediators released by activated microglia contributes to neuron damage in neurodegenerative disease. Therefore, identification of a compound that has anti-inflammatory activities and inhibits microglial activation may be an alternative therapeutic approach for the treatment of neurodegenerative diseases. Cyanidin-3-O-glucoside (C3G), a type of anthocyanin, possesses powerful anti-inflammatory activities. In this study, the anti-inflammatory effects of C3G were investigated in LPS-stimulated BV2 microglia. The results indicate that pretreatment with C3G significantly suppresses microglial activation and the production of neurotoxic mediators including nitric oxide (NO), prostaglandin E2 (PGE2), and pro-inflammatory cytokines such as interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) in LPS-activated BV2 cells. Moreover, C3G downregulates the gene expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokines via the suppression of NF-κB and p38 MAPK signaling pathways. Furthermore, a co-culture system to determine the indirect neuroprotective effects of C3G was used. Results demonstrated that conditioned medium (CM) from LPS-stimulated BV2 cells can promote the apoptosis of differentiated pheochromocytoma (PC12) cells through the activation of caspase-3, while C3G pretreatment in BV2 microglia can protect differentiated PC12 cells from microglial activation-induced apoptosis. Therefore, C3G may be a potential therapeutic agent for the treatment and prevention of neurodegenerative diseases associated with microglial activation.


Assuntos
Antocianinas/farmacologia , Apoptose/efeitos dos fármacos , Glucosídeos/farmacologia , Microglia/efeitos dos fármacos , Animais , Caspase 3/metabolismo , Técnicas de Cocultura , Meios de Cultivo Condicionados/farmacologia , Ciclo-Oxigenase 2/biossíntese , Dinoprostona/metabolismo , Regulação para Baixo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos , Camundongos , Microglia/metabolismo , NF-kappa B/metabolismo , Fármacos Neuroprotetores/farmacologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/biossíntese , Células PC12 , Ratos , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
9.
Cell Biochem Biophys ; 76(1-2): 279-292, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28831668

RESUMO

Osteoarthritis is a degenerative joint disease in which interleukin-1ß plays a major role in the inflammatory process. Administration of collagen hydrolysate was an optional treatment of osteoarthritis. Fish has become an interesting source of collagen hydrolysate because of religious reason and there is no risk from mad cow disease. However, the effects of different sizes of fish collagen hydrolysate on cartilage and chondrocyte metabolism have not been well studied yet. This study examined the effect of different sizes of fish collagen hydrolysate on cartilage metabolism. Three different sizes of fish collagen hydrolysate were prepared by size exclusion using centrifugation, which composed of small fraction (<3 kDa), medium fraction (3-10 kDa) and large fraction (>10 kDa). Using porcine cartilage explant, in physiological condition, all the three fractions had no effect on cartilage metabolism, but they could induce pro-MMP3 and pro-MMP13 secretions through activation of p-ERK and p-p38. In pathological condition induced by interleukin-1ß and oncostatin-M, small and medium fractions showed additive effect with interleukin-1ß and oncostatin-M on cartilage degradation, whereas large size had no effect. In addition, the effect of small size occurred through further activation of p-p65, which resulted in further induction of active-MMP13, while medium size had a different mechanism. In conclusion, all three fractions fish collagen hydrolysate had no effect on cartilage metabolism in physiological condition, but small and medium fractions had adverse effect on cartilage in pathological condition. Taken together, various sizes of fish collagen hydrolysate showed different effects on cartilage metabolism. Therefore, different sizes of fish collagen hydrolysates play different roles on cartilage metabolism, especially in the pathological condition.


Assuntos
Cartilagem Articular/efeitos dos fármacos , Colágeno/farmacologia , Proteínas de Peixes/farmacologia , Peixes/metabolismo , Animais , Cartilagem Articular/metabolismo , Células Cultivadas , Condrócitos/citologia , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Colágeno/química , Colágeno/isolamento & purificação , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Proteínas de Peixes/química , Proteínas de Peixes/isolamento & purificação , Humanos , Técnicas In Vitro , Interleucina-1beta/metabolismo , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , Oncostatina M/metabolismo , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Transdução de Sinais/efeitos dos fármacos , Suínos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
10.
Connect Tissue Res ; 59(4): 316-331, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-28956662

RESUMO

Osteoarthritis (OA) is the most common form of arthritis. Obesity has been believed to be an important risk factor for OA development and the progression of not only load-bearing joints, but low-load-bearing joints as well. Increased leptin has been the focus of a link between obesity and OA. In this study, the effects of pathological (100ng/ml) or supra-pathological (10µg/ml) concentrations of leptin alone or in combination with IL1ß on cartilage metabolisms were studied in porcine cartilage explant. The involved mechanisms were examined in human articular chondrocytes (HACs). Moreover, the protective effect of omega-3 polyunsaturated acids, eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) was also investigated. Leptin (10µg/ml) alone or in combination with IL1ß could induce cartilage destruction, although lower concentrations had no effect. Leptin activated NFκB, ERK, JNK and p38 in HACs, which led to the induction of MMP3, MMP13 and ADAMTS4 secretions. The combined effect could further induce those enzymes through the additive effect on activation of NFκB and JNK. Interestingly, both EPA and DHA could inhibit cartilage damage induced by leptin plus IL1ß by reducing the activation of NFκB and JNK, which led to the decrease of ADAMTS4 secretion. Altogether, only a supra-pathological concentration of leptin alone or in combination with IL1ß could induce cartilage destruction, whereas a pathological one could not. This effect could be inhibited by EPA and DHA. To gain greater understanding of the link between leptin and OA, the effect of different levels of leptin on several states of OA cartilage requires further investigation.


Assuntos
Cartilagem Articular/patologia , Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Interleucina-1beta/efeitos adversos , Leptina/efeitos adversos , Proteína ADAMTS4/metabolismo , Animais , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Ativação Enzimática/efeitos dos fármacos , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Suínos
11.
BMC Complement Altern Med ; 17(1): 532, 2017 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-29237438

RESUMO

BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disease that causes chronic synovitis, cartilage degradation and bone deformities. Synovitis is the term for inflammation of the synovial membrane, an early stage of RA. The pathogenesis of the disease occurs through cytokine induction. The major cytokine that increases the severity of RA is TNF-α. Thus, inhibition of the TNF-α cascade is an effective way to diminish the progression of the disease. We are interested in investigating the difference between primary human synovial fibroblast (hSF) cells and SW982 as synovitis models induced by TNF-α and in monitoring their responses to sesamin as an anti-inflammatory phytochemical. METHOD: The designed experiments were performed in hSF cells or the SW982 cell line treated with 10 ng/ml TNF-α with or without 0.25, 0.5 or 1 µM sesamin. Subsequently, pro-inflammatory cytokine genes and proteins were measured in parallel with a study of associated signalling transduction involved in inflammatory processes, including NF-κB and MAPK pathways. RESULTS: The results demonstrated that although hSF and SW982 cells responded to TNF-α induction in the same fashion, they reacted at different levels. TNF-α could induce IL-6, IL-8 and IL-1ß in both cell types, but the levels in SW982 cells were much higher than in hSF cells. This characteristic was due to the different induction of MAPKs in each cell type. Both cell types reacted to sesamin in almost the same fashion. However, hSF cells were more sensitive to sesamin than SW982 cells in terms of the anti-RA effect. CONCLUSIONS: The responses of TNF-α-induced hSF and SW982 were different at the signal transduction level. However, the two cell types showed almost the same reaction to sesamin treatment in terms of the end point of the response.


Assuntos
Dioxóis/farmacologia , Fibroblastos/efeitos dos fármacos , Lignanas/farmacologia , Membrana Sinovial/citologia , Sinovite/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Artrite Reumatoide , Linhagem Celular , Citocinas/metabolismo , Humanos , Modelos Biológicos , Transdução de Sinais/efeitos dos fármacos
12.
Arch Pharm Res ; 40(10): 1186-1196, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28861734

RESUMO

The promotional nature of sesamin on human osteoblast differentiation has been proven. Here, the effect of sesamin on human osteoclasts was investigated in terms of differentiation and function in M-CSF and RANKL induced human PBMCs. Sesamin treatment significantly decreased the number of differentiated osteoclasts observed by TRAP staining; however, sesamin inhibition did not result from the alteration of precursor cell proliferation. Sesamin did not decrease NFATc1 gene expression, which opposed the decreasing trend of CathK and TRAP expression. DC-STAMP, but not Atp6v0d2, also significantly decreased in the presence of 14 µM sesamin. Expressions of CCR2b and CCR4 as chemokine receptors were significantly down-regulated. Sesamin might mediate the inhibition of human osteoclast differentiation, the recruitment of precursor cells and F-actin formation. Decrease in the area of the resorption pits and the collagen released from the bone slices under sesamin treatment emphasized the inhibitory effects on both the differentiation and function of osteoclasts. Sesamin is a promising phytochemical agent inhibiting osteoclast differentiation and function.


Assuntos
Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Dioxóis/farmacologia , Lignanas/farmacologia , Osteoclastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Células da Medula Óssea/citologia , Reabsorção Óssea/metabolismo , Reabsorção Óssea/prevenção & controle , Técnicas de Cultura de Células , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Expressão Gênica/efeitos dos fármacos , Voluntários Saudáveis , Humanos , Fator Estimulador de Colônias de Macrófagos/metabolismo , Osteoclastos/citologia , Ligante RANK/metabolismo
13.
Asian Pac J Trop Med ; 10(3): 238-252, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28442107

RESUMO

OBJECTIVE: To investigate the biological effects of the Mucuna pruriens (M. pruriens) seed extracts that lacked l-DOPA, which was formerly reported as the active ingredient, on erectile dysfunction (ED) both in vitro and in vivo. METHODS: Seed of M. pruriens plant that cultivated in Mae Taeng District, Chiang Mai Province, Thailand, was collected. Component of its seeds were extracted and isolated into 2 fractions using methanol, polar and nonpolar. Each fraction was investigated for phytochemicals using gas chromatography and mass spectroscopy and was screened for biological activity in vitro using three different cell lines. The most biological active fraction was used to treat both streptozotocin (STZ)-induced diabetes mellitus-erectile dysfunction (DM-ED) male Wistar rats and normal rats (n = 6 per groups) to compare the effect on sexual behavior parameters, including number of intromission, mounting and ejaculation, with that of rats given Sildenafil by individually pairing with their female counterparts. Penile tissues and serums were collected to determine histological structure, related gene expression and biomolecules. RESULTS: The phytochemicals of the polar fraction were possibly catechol and its derivatives plus polyphenols, whereas the nonpolar fraction consisted of lipid derivatives. l-DOPA was not detected in either of the extracts. The polar fraction was able to up-regulate the expression of ED-related genes including eNOS and nNOS in vitro which subsequently promotes nitric oxide production and maintains intracellular cyclic guanosine monophosphate levels. When administrated to DM-ED rats, the polar extract significantly improved all sexual behavior parameters in DM-ED rats compared to untreated group (18.3 ± 1.8 to 10.8 ± 2.9 for intromission, 9.8 ± 2.2 to 5.7 ± 1.3 for mounting, and 1.8 ± 0.6 to 0.2 ± 0.4 for ejaculation). That effect might due to the ability of the extract to stimulate the expression of eNOS and nNOS which results in nitric oxide production and subsequently maintains cyclic guanosine monophosphate levels in penile tissue. Moreover, this extract may also prevent penile tissue deterioration due to diabetes. CONCLUSIONS: The polar extract of M. pruriens seed can be used for ED therapy, especially in patients with metabolic diseases including diabetes. The action of the extract might be due to catechol and its derivatives and polyphenols.

14.
BMC Complement Altern Med ; 16: 205, 2016 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-27400672

RESUMO

BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disease associated with chronic inflammatory arthritis. TNF-α and OSM are pro-inflammatory cytokines that play a key role in RA progression. Thus, reducing the effects of both cytokines is practical in order to relieve the progression of the disease. This current study is interested in sesamin, an active compound in sesame seeds. Sesamin has been shown to be a chondroprotective agent in osteoarthritis models. Here, we have evaluated a porcine cartilage explant as a cartilage degradation model related to RA induced by TNF-α and/or OSM in order to investigate the effects of sesamin on TNF-α and OSM in the cartilage degradation model. METHODS: A porcine cartilage explant was induced with a combination of TNF-α and OSM (test group) or IL-1ß and OSM (control group) followed by a co-treatment of sesamin over a long-term period (35 days). After which, the tested explants were analyzed for indications of both the remaining and the degradation aspects using glycosaminoglycan and collagen as an indicator. RESULTS: The combination of TNF-α and OSM promoted cartilage degradation more than either TNF-α or OSM alone and was comparable with the combination of IL-1ß and OSM. Sesamin could be offering protection against cartilage degradation by reducing GAGs and collagen turnover in the generated model. CONCLUSIONS: Sesamin might be a promising agent as an alternative treatment for RA patients. Furthermore, the generated model revealed itself to be an impressive test model for the analysis of phytochemical substances against the cartilage degradation model for RA. The model could be used to test for the prevention of cartilage degradation in other biological agents induced with TNF-α and OSM as well.


Assuntos
Cartilagem/efeitos dos fármacos , Dioxóis/farmacologia , Lignanas/farmacologia , Oncostatina M/metabolismo , Substâncias Protetoras/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Artrite Reumatoide , Cartilagem/metabolismo , Dioxóis/química , Imuno-Histoquímica , Lignanas/química , Modelos Biológicos , Substâncias Protetoras/química , Suínos
15.
BMC Biotechnol ; 14: 108, 2014 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-25543823

RESUMO

BACKGROUND: Cartilage tissue engineering offers new strategies in repairing damaged cartilage. Scaffolds have been used for the in vitro and in vivo procedures for this application, which demonstrates the compatible biological and physical properties that mimic natural tissues. Several types of scaffolds were used and had different effects on cell functions. The study was designed to develop a functional gelatin scaffold by adsorption of hyaluronan (HA) and the transforming growth factor ß3 (TGF-ß3) in a commercially available gelatin scaffold. RESULTS: The biological properties of human articular chondrocytes were investigated during a 21-day cultivation embedded in either HA + TGF-ß3 adsorbed scaffolds or the conventional supplemented method. The rising of proliferation of chondrocytes embedded in adsorbed scaffolds was observed at day 17 and 21 of cultivation (1.27 and 1.28 fold, respectively). The chondrogenic gene expression of the chondrocytes embedded in HA + TGF-ß3 adsorbed scaffolds significantly increased: SOX-9 (1.65 fold), ACAN (7.65 fold) and COL2A1 (1.83 fold). Remarkably, over the 21 days of cultivation, HA + TGF-ß3 adsorbed scaffolds promoted the extracellular matrix molecules production with higher accumulation of HA (1.2 fold), collagen (1.42 fold) and uronic acid (1.41 fold). Moreover, the cell population and extracellular matrix production, which were examined by a histological analysis and a scanning electron microscope, were correlated with the biochemical analysis. CONCLUSION: A small amount of HA and TGF-ß3 initially adsorbed in the scaffolds (70 µg and 10 ng, respectively) was consumed over the 21-day cultivation. The HA + TGF-ß3 adsorbed gelatin scaffold is effective and more suitable than the conventional supplemented method for the in vitro assessment of human chondrocyte 3D culture.


Assuntos
Cartilagem Articular/citologia , Condrócitos/citologia , Ácido Hialurônico/metabolismo , Tecidos Suporte/química , Fator de Crescimento Transformador beta3/metabolismo , Adsorção , Cartilagem Articular/metabolismo , Proliferação de Células , Condrócitos/metabolismo , Humanos , Engenharia Tecidual/instrumentação
16.
Phytochemistry ; 80: 77-88, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22704650

RESUMO

Osteoarthritis (OA) is a major disability of elderly people. Sesamin is the main compound in Sesamun indicum Linn., and it has an anti-inflammatory effect by specifically inhibiting Δ5-desaturase in polyunsaturated fatty acid biosynthesis. The chondroprotective effects of sesamin were thus studied in a porcine cartilage explant induced with interleukin-1beta (IL-1ß) and in a papain-induced osteoarthritis rat model. With the porcine cartilage explant, IL-1ß induced release of sulfated-glycosaminoglycan (s-GAG) and hydroxyproline release, and this induction was significantly inhibited by sesamin. This ability to inhibit these processes might be due to its ability to decrease expression of MMP-1, -3 and -13, which can degrade both PGs and type II collagen, both at the mRNA and protein levels. Interestingly, activation of MMP-3 might also be inhibited by sesamin. Moreover, in human articular chondrocytes (HACs), some pathways of IL-1ß signal transduction were inhibited by sesamin: p38 and JNK. In the papain-induced OA rat model, sesamin treatment reversed the following pathological changes in OA cartilage: reduced disorganization of chondrocytes in cartilage, increased cartilage thickness, and decreased type II collagen and PGs loss. Sesamin alone might increase formation of type II collagen and PGs in the cartilage tissue of control rats. These results demonstrate that sesamin efficiently suppressed the pathological processes in an OA model. Thus, sesamin could be a potential therapeutic strategy for treatment of OA.


Assuntos
Anti-Inflamatórios/farmacologia , Condrócitos/efeitos dos fármacos , Dioxóis/farmacologia , Lignanas/farmacologia , Animais , Anti-Inflamatórios/uso terapêutico , Cartilagem/citologia , Cartilagem/patologia , Condrócitos/metabolismo , Condrócitos/patologia , Colágeno Tipo II/metabolismo , Dioxóis/uso terapêutico , Relação Dose-Resposta a Droga , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Glicosaminoglicanos/metabolismo , Humanos , Hidroxiprolina/metabolismo , Interleucina-1beta/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Lignanas/uso terapêutico , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Metaloproteinases da Matriz/metabolismo , Osteoartrite/induzido quimicamente , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Osteoartrite/patologia , Papaína/efeitos adversos , Fosforilação/efeitos dos fármacos , Proteoglicanas/metabolismo , Ratos , Suínos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
17.
In Vitro Cell Dev Biol Anim ; 48(1): 43-53, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22179678

RESUMO

Interleukin-1ß (IL-1ß) induces the expression of matrix metalloproteinases (MMPs) implicated in cartilage and joint degradation in osteoarthritis (OA) and rheumatoid arthritis (RA). Polyoxypregnane glycoside (PPG), active compound was identified from Dregea volubilis extract by chemical analysis, shown to exert chondroprotective effects in cartilage explant models. However, no studies have been undertaken for the molecular investigation of whether PPG constituents protect the human articular chondrocyte (HAC). In the present studies, HAC was co-treated with IL-1ß and PPG. The expression of MMPs, type II collagen, phosphorylation of mitogen-activated protein kinases (MAPKs) and NF-κB signaling pathway were determined by Western immunoblotting. PPG (6.25-25 µM) decreased the IL-1ß-induced HA release from chondrocyte to culture medium. The mode of action of PPG was likely mediated through inhibiting expression of MMP-1, -3 and -13 in the medium, which was associated with the inhibition of mRNA expression. PPG had no effect on IL-1ß-induced phosphorylation of MAPK pathway. Conversely, PPG decreased phosphorylation of IκB kinase and IκBα degradation. Taken together, these results indicate that PPG may inhibit cartilage degradation in OA and may also be used as nutritional supplement for maintaining joint integrity and function.


Assuntos
Condrócitos/metabolismo , Interleucina-1beta/antagonistas & inibidores , Metaloproteinases da Matriz/metabolismo , NF-kappa B/metabolismo , Extratos Vegetais/farmacologia , Animais , Apocynaceae/química , Células Cultivadas , Condrócitos/citologia , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Glicosídeos/farmacologia , Humanos , Interleucina-1beta/administração & dosagem , Metaloproteinases da Matriz/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , NF-kappa B/genética , Fosforilação/efeitos dos fármacos , Extratos Vegetais/química , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Suínos
18.
In Vitro Cell Dev Biol Anim ; 47(3): 183-7, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21132464

RESUMO

Alpinia galanga has been used as alternative medicine for anti-rheumatic activities. However, the precise action of the extract on arthritic diseases is not yet fully understood. In this study, we investigated the effects of A. galanga extracts on the expression of genes involved in catabolic activities in an interleukin-1ß (IL-1ß)-induced human synovial fibroblast as an inflammatory model. Confluent primary human synovial fibroblasts were treated for 24 h with A. galanga hexane extracts in the presence of recombinant human IL-1ß. MMPs in the culture medium were monitored by gelatin zymography. Total RNA was isolated from the cell lysate and analyzed via semi-quantitative RT-PCR. After treatment with A. galanga extracts, MMP-2 activity in the culture medium was significantly reduced. In addition, MMP-1, MMP-3, MMP-13, and Cox-2 expression were downregulated. These data suggest that the decrease of gene expression and production of MMPs in synovial fibroblasts against inflammatory stimuli could be due to the effects of the A. galanga extracts. Therefore, A. galanga extracts might be a promising therapeutic agent for arthritis.


Assuntos
Alpinia/química , Regulação para Baixo/efeitos dos fármacos , Fibroblastos/enzimologia , Interleucina-1beta/farmacologia , Metaloproteinases da Matriz/metabolismo , Extratos Vegetais/farmacologia , Membrana Sinovial/citologia , Cromatografia Líquida de Alta Pressão , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Fibroblastos/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Metaloproteinases da Matriz/genética
19.
BMC Musculoskelet Disord ; 11: 162, 2010 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-20630114

RESUMO

BACKGROUND: Glucosamine (GlcN) is a well-recognized candidate for treatment of osteoarthritis. However, it is currently used in derivative forms, such as glucosamine-hydrochloride (GlcN-HCl) or glucosamine sulfate (GlcN-S). However, the molecular mode of action remains unclear. In this study, we compared the effects of Glucose (Glc), Glucuronic acid (GlcA), Glucosamine hydrochloride (GlcN-HCl) and Glucosamine sulfate (GlcN-S) on cartilage degradation. METHODS: Porcine cartilage explants were co-cultured with recombinant human IL-1beta and each tested substance for 3 days. HA, s-GAG and MMP-2 releases to media were measured using ELISA, dye-binding assay and gelatin zymography, respectively. Similar studies were performed in a human articular chondrocytes (HAC) monolayer culture, where cells were co-treated with IL-1beta and each reagent for 24 hours. Subsequently, cells were harvested and gene expression measured using RT-PCR. All experiments were carried out in triplicate. Student's t-tests were used for statistical analysis. RESULTS: In cartilage explants treated with IL-1beta, GlcN-S had the highest chondroprotective activity of all four chemicals as shown by the inhibition of HA, s-GAG and MMP-2 released from cartilage. The anabolic (aggrecan core protein; AGG, SOX9) and catabolic (MMP-3, -13) genes in HACs treated with IL-1beta and with/without chemicals were studied using RT-PCR. It was found that, GlcN-HCl and GlcN-S could reduce the expression of both MMP-3 and -13 genes. The IL-1beta induced-MMP-13 gene expression was decreased maximally by GlcN-S, while the reduction of induced-MMP-3 gene expression was greatest with GlcN-HCl. Glc and GlcA reversed the effect of IL-1beta on the expression of AGG and SOX9, but other substances had no effect. CONCLUSION: This study shows that glucosamine derivatives can alter anabolic and catabolic processes in HACs induced by IL-1beta. GlcN-S and GluN-HCl decreased induced MMP-3 and -13 expressions, while Glc and GlcA increased reduced-AGG and SOX9 expression. The chondroprotective study using porcine cartilage explant showed that GlcN-S had the strongest effect.


Assuntos
Cartilagem Articular/efeitos dos fármacos , Glucosamina/química , Glucosamina/farmacologia , Glucose/análogos & derivados , Glucose/farmacologia , Agrecanas/metabolismo , Animais , Doenças das Cartilagens/tratamento farmacológico , Doenças das Cartilagens/metabolismo , Doenças das Cartilagens/patologia , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Células Cultivadas , Técnicas de Cocultura , Glucosamina/uso terapêutico , Glucose/uso terapêutico , Ácido Glucurônico/farmacologia , Ácido Glucurônico/uso terapêutico , Glicosaminoglicanos/metabolismo , Humanos , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/fisiologia , Interleucina-1beta/antagonistas & inibidores , Interleucina-1beta/fisiologia , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Osteoartrite/patologia , Fatores de Transcrição SOX9/metabolismo , Sus scrofa
20.
Phytochemistry ; 70(2): 237-43, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19118849

RESUMO

Osteoarthritis (OA) is the most common form of arthritis and affects millions of people worldwide. Patients have traditionally been treated with non-steroidal anti-inflammatory drugs (NSAIDs), but these are associated with significant side effects. Purification of the acetone extract of Alpinia galanga afforded p-hydroxycinnamaldehyde, as identified by nuclear magnetic resonance and mass spectrometry analyses. By exploiting the cartilage explant culture, p-hydroxycinnamaldehyde suppressed loss of uronic acid, resulting in release of hyaluronan (HA), sulfated glycosaminoglycans (s-GAGs) and matrix metalloproteinases (MMPs). p-Hydroxycinnamaldehyde and interleukin-1beta (IL-1beta), when incubated in primary human chondrocytes, also reduced release of HA, s-GAG and MMP-2. The results demonstrated: (a) that expression levels of the catabolic genes MMP-3 and MMP-13 were suppressed and (b) mRNA expression levels of anabolic genes of collagen II, SOX9 and aggrecan were increased. This study shows that p-hydroxycinnaldehyde from A. galanga Linn. is a potential therapeutic agent for treatment of OA.


Assuntos
Alpinia/química , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Cinamatos/química , Cinamatos/farmacologia , Extratos Vegetais/química , Acetona , Animais , Cartilagem/efeitos dos fármacos , Cartilagem/cirurgia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Estrutura Molecular , Suínos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...