RESUMO
Valproic acid (VPA) has been reported to inhibit cancer cell growth and has therapeutic use in retinal diseases. However, the mechanism of this action remains unclear. In order to explore this mechanism, primary human retinal pigment epithelial (hRPE) cell cultures were established. Cell viability was assessed by the trypan blue exclusion method (T), and the cell proliferation was measured by 3H-thymidine incorporation (3H-thy). P38 synthesis was quantitated by using 14C-methionine-labeled P38 (14C-P38) by using P38-specific antibody. SB203580 (SB), a selective inhibitor of p38 MAPK, was also used to test the specificity of P38 stimulation. Antinuclear staining (NS) studies were performed by DAPI. Statistical significance was established by student's t-test. We observed that VPA (1 mM) inhibited 10% fetal bovine serum (FBS)-stimulated cell proliferation (1.75 ± 0.37 vs. 3.25 ± 0.68 cells per 1 µl ± SEM, p < 0.05, n = 4). VPA also stimulated 14C-P38 synthesis in a dose-dependent manner. SB (30 µM) inhibited VPA (4 mM)-stimulated 14C-P38 synthesis (197.74 ± 41.17 vs. 425.89 ± 59.17, CPM ± SEM, p < 0.05, n = 4) and increased hRPE cell proliferation (1.79 ± 0.45 vs. 4.93 ± 1.12 cells per 1 µl ± SEM, p < 0.05, n = 4); NS demonstrated VPA-induced cell damage. We conclude that VPA inhibits hRPE cell growth via P38 MAP mechanism and may be of therapeutic value in treating or preventing proliferative eye diseases.
Assuntos
Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Epitélio Pigmentado da Retina/efeitos dos fármacos , Ácido Valproico/farmacologia , Núcleo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Indução Enzimática/efeitos dos fármacos , Humanos , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
AIMS: Muscle mass is regulated by the balance between the synthesis and degradation of muscle proteins. Loss of skeletal muscle mass is associated with an increased risk of developing metabolic diseases such as obesity and type 2 diabetes mellitus. The aim of this study was to clarify the effects of licorice flavonoid oil on muscle mass in KK-Ay/Ta mice. MAIN METHODS: Male genetically type II diabetic KK-Ay/Ta mice received 0, 1, or 1.5â¯g/kg BW of licorice flavonoid oil by mouth once daily for 4â¯weeks. After 4â¯weeks, the femoral and soleus muscles were collected for western blotting for evaluation of the mTOR/p70 S6K, p38/FoxO3a, and Akt/FoxO3a signaling pathways. KEY FINDINGS: Ingestion of licorice flavonoid oil significantly enhanced femoral muscle mass without affecting body weight in KK-Ay/Ta mice. Licorice flavonoid oil also decreased expression of MuRF1 and atrogin-1, which are both markers of muscle atrophy. The mechanisms by which licorice flavonoid oil enhances muscle mass include activation of mTOR and p70 S6K, and regulation of phosphorylation of FoxO3a. SIGNIFICANCE: Ingestion of licorice flavonoids may help to prevent muscle atrophy.
Assuntos
Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Flavonoides/farmacologia , Glycyrrhiza/química , Músculo Esquelético/efeitos dos fármacos , Óleos de Plantas/farmacologia , Animais , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/biossíntese , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Proteína Forkhead Box O3/biossíntese , Proteína Forkhead Box O3/genética , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Camundongos , Proteínas Musculares/antagonistas & inibidores , Proteínas Musculares/biossíntese , Proteínas Musculares/genética , Músculo Esquelético/crescimento & desenvolvimento , Músculo Esquelético/patologia , Tamanho do Órgão/efeitos dos fármacos , Proteínas Ligases SKP Culina F-Box/antagonistas & inibidores , Proteínas Ligases SKP Culina F-Box/biossíntese , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/biossíntese , Proteínas com Motivo Tripartido , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
The triterpenoid, bauerenol, from Suregada angustifolia (Baill. ex Muell.-Arg.) Airy Shaw (Euphorbiaceae) was screened for anti-cancer property using hepatocellular carcinoma cell line, HepG2. Bauerenol exhibited growth inhibitory and apoptosis inducing potential against HepG2 cancer cells. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytotoxic assay revealed that bauerenol treatment significantly reduced the growth of HepG2 cells in a time- and dose-dependent manner with 50% growth inhibitory concentration doses of 45 and 25 µg/mL at 24 and 48 h treatments, respectively. Bauerenol-induced cell death reflected apoptotic morphological features, that is, cell membrane blebbing, vacuolization, chromatin condensation, and nuclear fragmentation. In addition, bauerenol treatment diminished the mitochondrial membrane potential, by inducing the efflux of cytochrome c, downregulating the levels of anti-apoptotic Bcl-2 as well as upregulating the levels of pro-apoptotic Bax, and inducing caspase activation and poly (ADP-ribose) polymerase cleavage. Moreover, bauerenol treatment activates p38MAPK and inactivates the anti-apoptotic kinases Akt and ERK1/2 through the induction of reactive oxygen species. Furthermore, bauerenol-mediated S-phase arrest was associated with downregulation of cell cycle-rate-limiting factor (cyclin D1) and upregulation of cyclin-dependent kinase inhibitor p21 and tumor suppressor p53. Interestingly, pre-treatment of cells with reactive oxygen species inhibitor and p38 inhibitor significantly decreases bauerenol-induced cytotoxicity, Bax upregulation, and p38 activation. This study clearly states that bauerenol induces cell cycle arrest and apoptosis through the reactive oxygen species-dependent p38MAPK activation in HepG2 cancer cells.
Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Triterpenos/administração & dosagem , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Ciclina D1/biossíntese , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Espécies Reativas de Oxigênio/metabolismo , Suregada/química , Triterpenos/química , Proteína X Associada a bcl-2/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Vascular smooth muscle cell (VSMC) proliferation is a key event in the development of instent restenosis. Evodiamine is an indole alkaloid extracted from the Chinese medicine, evodia, and has been shown to inhibit tumor cell proliferation and protect the cardiovascular system. However, whether evodiamine affects VSMC proliferation remains to be elucidated. Therefore, the present study examined the effects and the mechanisms of action of evodiamine on the proliferation of rat VSMCs. The cells were treated with evodiamine alone or in combination with plateletderived growth factorBB (PDGFBB) stimulation. It was found that evodiamine inhibited PDGFBBinduced VSMC proliferation in a dosedependent manner, without inducing cell death. Evodiamine also retarded cell cycle progression, evidenced by the suppression of the expression of cell cyclepromoting cyclin proteins and cyclindependent kinases. In addition, evodiamine attenuated the PDGFBBinduced phosphorylation of mitogenactivated protein kinases p38 and extracellular signalregulated kinases 1/2, however, it had no effect on the phosphorylation of Akt. Evodiamine also inhibited the increase of reactive oxygen species generation and upregulated the mRNA expression levels of genes encoding antioxidant enzymes. These findings provide important insights into the mechanisms underlying the vasoprotective actions of evodiamine and suggest that it may be a useful therapeutic agent for the treatment of vascular occlusive disease.
Assuntos
Proliferação de Células/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Quinazolinas/administração & dosagem , Animais , Ciclo Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Evodia/química , Regulação da Expressão Gênica/efeitos dos fármacos , Medicina Tradicional Chinesa , Músculo Liso Vascular/crescimento & desenvolvimento , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-sis/administração & dosagem , Proteínas Proto-Oncogênicas c-sis/metabolismo , Ratos , Espécies Reativas de Oxigênio/metabolismo , Doenças Vasculares/tratamento farmacológico , Doenças Vasculares/genética , Doenças Vasculares/patologia , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
The nuclear factor-κB (NF-κB) transcription factors control many physiological processes including inflammation, immunity, apoptosis and angiogenesis. In our search for NF-κB inhibitors from natural resources, we identified baicalein from Scutellaria baicalensis as an inhibitor of NF-κB activation. As examined by the NF-κB luciferase reporter assay, we found that baicalein suppressed TNF-α-induced NF-κB activation in a dose-dependent manner. It also inhibited TNF-α-induced nuclear translocation of p65 through inhibition of phosphorylation and degradation of IκBα. Furthermore, baicalein blocked the TNF-α-induced expression of NF-κB target genes involved in anti-apoptosis (cIAP-1, cIAP-2, FLIP and BCL-2), proliferation (COX-2, cyclin D1 and c-Myc), invasion (MMP9), angiogenesis (VEGF) and major inflammatory cytokines (IL-8 and MCP1). The flow cytometric analysis indicated that baicalein potentiated TNF-α-induced apoptosis and induced G1 phase arrest in HeLa cells. Moreover, baicalein significantly blocked activation of p38, extracellular signal-regulated kinase 1/2 (ERK1/2). Our results imply that baicalein could be a lead compound for the modulation of inflammatory diseases as well as certain cancers in which inhibition of NF-κB activity may be desirable.
Assuntos
Flavanonas/administração & dosagem , Extratos Vegetais/administração & dosagem , Fator de Transcrição RelA/biossíntese , Fator de Necrose Tumoral alfa/genética , Apoptose/genética , Proliferação de Células/efeitos dos fármacos , Citometria de Fluxo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células HeLa , Humanos , Quinase I-kappa B/biossíntese , Quinase I-kappa B/genética , NF-kappa B/biossíntese , NF-kappa B/genética , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , Fosforilação , Scutellaria baicalensis , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/genética , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Sulfur mustard (SM) is an alkylating agent, which has been used as in chemical warfare in a number of conflicts. As the generation of reactive oxygen species (ROS), and adducts in DNA and proteins have been suggested as the mechanism underlying SMinduced cytotoxicity, the present study screened several antioxidant candidates, including tannic acid, deferoxamine mesylate, trolox, vitamin C, ellagic acid and caffeic acid (CA) to assess their potential as therapeutic agents for SMinduced cell death. Among several antioxidants, CA partially alleviated SMinduced cell death in a dosedependent manner. Although CA treatment decreased the phosphorylation of p38 mitogenactivated protein (MAP) kinase and p53, p38 MAP kinase inhibition by SB203580 did not affect SMinduced cell death. As CA has also been reported as a 15lipoxygenase (15LOX) inhibitor, the role of 15LOX in SMinduced cytotoxicity was also examined. Similar to the results observed with CA, treatment with PD146176, a specific 15LOX inhibitor, decreased SMinduced cytotoxicity, accompanied by decreases in the production of tumor necrosis factorα and 15hydroxyeicosatetraenoic acid. Furthermore, the present study investigated the protective effects of two natural 15LOX inhibitors, morin hydrate and quercetin, in SMinduced cytotoxicity. As expected, these inhibitors had similar protective effects against SMinduced cytotoxicity. These antioxidants also reduced the generation of ROS and nitrate/nitrite. Therefore, the results of the present study indicated that the natural products, CA, quercetin and morin hydrate, offer potential as adjuvant therapeutic agents for SMinduced toxicity, not only by reducing inflammation mediated by the p38 and LOX signaling pathways, but also by decreasing the generation of ROS and nitrate/nitrite.
Assuntos
Ácidos Cafeicos/administração & dosagem , Morte Celular/efeitos dos fármacos , Flavonoides/administração & dosagem , Lipoxigenase/genética , Quercetina/administração & dosagem , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Antioxidantes/administração & dosagem , Adutos de DNA/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Imidazóis/administração & dosagem , Queratinócitos/efeitos dos fármacos , Lipoxigenase/biossíntese , Gás de Mostarda/toxicidade , Fosforilação , Piridinas/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Proteína Supressora de Tumor p53/biossíntese , Proteína Supressora de Tumor p53/genética , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Tenuigenin (TEN), the main active component of Polygala tenuifolia, has been reported to have anti-inflammatory effects. However, the effects of TEN on IL-1ß-stimulated osteoarthritis chondrocytes have not been reported. The purpose of this study was to investigate the anti-inflammatory effects and mechanism of TEN on IL-1ß-stimulated human osteoarthritis chondrocytes. Human osteoarthritis chondrocytes were pretreated with or without TEN for 1 h and then stimulated with IL-1ß. The production of NO and PGE2 were detected by the Griess reagent and ELISA. The expression of NF-κB and MAPKs (p38, JNK, ERK) were measured by Western blot analysis. The production of MMP-1, MMP3, and MMP13 were measured by ELISA. The results showed that treatment of TEN significantly inhibited IL-1ß-induced NO and PGE2 production. TEN also suppressed IL-1ß-induced MMP-1, MMP3, and MMP13 expression. Furthermore, TEN was found to inhibit IL-1ß-induced NF-κB activation, PI3K, and AKT phosphorylation. In conclusion, these results suggest that TEN inhibits IL-1ß-induced inflammation in human osteoarthritis chondrocytes by inhibiting PI3K/AKT/NF-κB signaling pathway.
Assuntos
Anti-Inflamatórios/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Inflamação/prevenção & controle , Interleucina-1beta/imunologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Transcrição RelA/metabolismo , Artroplastia do Joelho , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Dinoprostona/biossíntese , MAP Quinases Reguladas por Sinal Extracelular/biossíntese , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/biossíntese , Metaloproteinase 1 da Matriz/biossíntese , Metaloproteinase 13 da Matriz/biossíntese , Metaloproteinase 3 da Matriz/biossíntese , Óxido Nítrico/biossíntese , Osteoartrite/tratamento farmacológico , Fosforilação/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
The pro-inflammatory cytokine interleukin-1ß (IL-1ß) is known to play a crucial role in the pathogenesis of osteoarthritis (OA) by stimulating several mediators that contribute to cartilage degradation. Mori folium, the leaves of Morus alba L., has long been used in traditional medicine to treat diabetes, protect the liver, and lower blood pressure; however, the role of Mori folium in OA is not yet fully understood. Therefore, in the present study, we investigated whether Mori folium water extract (MF) inhibited the catabolic effects of IL-1ß in vitro, and also whether it inhibited the matrix metalloproteinases (MMPs), inducible nitric oxide (NO) synthase (iNOS) and cyclooxygenase-2 (COX-2) through the attenuation of nuclear factor-κB (NF-κB) and mitogen activated protein kinase (MAPK) pathways in SW1353 human chondrocytes. MMP proteins in culture medium were determined using a cytokinespecific enzyme-linked immunosorbent assay (ELISA). The production of NO and prostaglandin E2 (PGE2) were evaluated using Griess reagent and ELISA. Subsequently, the mRNA and protein levels of MMPs, iNOS, COX-2, NF-κB and MAPKs were examined by RT-qPCR and/or western blot analysis. The results indicate that MF significantly reduced the IL-1ßinduced release of MMP-1 and -13 in SW1353 cells, which was associated with the inhibition of MMP-1 and -13 mRNA and protein expression in a concentrationdependent manner at concentrations with no cytotoxicity. MF also attenuated the IL-1ß-induced production of NO and PGE2, and reduced iNOS and COX-2 expression. Furthermore, we noted that MF markedly suppressed the IL-1ßinduced nuclear translocation of NF-κB, which correlated with the inhibitory effects of MF on inhibitor-κB (IκB) degradation, and the phosphorylation of p38 MAPK was selectively restored by MF upon IL-1ß stimulation. These results indicate that MF inhibited the production and expression of MMP-1 and -13 and inflammatory mediators, at least in part, through suppressing the activation of either NF-κB or p38 MAPK in IL-1ß-treated SW1353 chondrocytes. Therefore, the novel findings of the present study suggest that MF is a potential therapeutic choice for chondroprotection against the collagen matrix breakdown in the cartilage of diseased tissues, such as those found in patients with arthritic disorders.
Assuntos
Condrócitos/efeitos dos fármacos , Inflamação/tratamento farmacológico , Interleucina-1beta/metabolismo , Osteoartrite/tratamento farmacológico , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Cartilagem/efeitos dos fármacos , Condrócitos/patologia , Ciclo-Oxigenase 2/biossíntese , Dinoprostona/biossíntese , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/genética , Inflamação/patologia , Metaloproteinases da Matriz/biossíntese , Morus/química , NF-kappa B/biossíntese , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/biossíntese , Osteoartrite/genética , Osteoartrite/patologia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/químicaRESUMO
Tanshinone IIA (Tan-IIA) is extracted from Danshen (Salviae miltiorrhizae radix). It possesses antitumor activity against a variety of human cancer cells and its induction of apoptosis and inhibition of proliferation of gastric cancer cells are well-documented. However, the molecular mechanisms by which Tan-IIA inhibits gastric cancer have not been well-elucidated. In the present study, we evaluated the cytotoxicity of Tan-IIA against human gastric cancer AGS cells by the (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) MTT assay. The protein expression of tumor necrosis factor-alpha (TNF-α), FAS, p53, p21, cyclin A, cyclin B1, extracellular-related kinase (ERK), phospho extracellular-related kinase (p-ERK), p38, p-p38, Jun-amino-terminal kinase (JNK), phospho Jun-amino-terminal kinase (p-JNK) and ß-actin in AGS cells were measured by western blotting. The cell-cycle distribution was analyzed by flow cytometry. The results showed that Tan-IIA inhibited AGS cells with time- and dose-dependent manners. AGS cells treated with Tan-IIA up-regulated the protein expression of TNFα, FAS, p-p38, p-JNK, p53, p21, caspase-3 and caspase-8 but reduced that of p-ERK, CDC2, cyclin A, and cyclin B1. The results also showed that Tan-IIA dose dependently induced G2/M phase arrest. These findings demonstrate that Tan-IIA can inhibit AGS human gastric cancer cells; one of the molecular mechanisms may be through increasing the protein expression of p-p38 and p-JNK, but decreasing that of p-ERK to induce the activation of p53, followed by increasing the protein expression of p21 to down-regulate CDC2 and cyclin B1 expression which then induces G2/M phase arrest. Another route may be through increasing the protein expression of TNF-α, FAS, caspase-8 and caspase-3 to induce apoptosis.
Assuntos
Abietanos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Neoplasias Gástricas/tratamento farmacológico , Apoptose/efeitos dos fármacos , Proteína Quinase CDC2 , Caspase 3/biossíntese , Caspase 8/biossíntese , Linhagem Celular Tumoral , Ciclina A/biossíntese , Ciclina B1/biossíntese , Quinases Ciclina-Dependentes/biossíntese , MAP Quinases Reguladas por Sinal Extracelular/biossíntese , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/biossíntese , Pontos de Checagem da Fase M do Ciclo Celular/efeitos dos fármacos , Extratos Vegetais/farmacologia , Proteína Supressora de Tumor p53/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
Botanicals are rich in bioactive compounds, and some offer numerous beneficial effects to animal and human health when consumed. It is well known that phytochemicals in cranberries have anti-oxidative and antimicrobial activities. Recently, an increasing body of evidence has demonstrated that cranberry phytochemicals may have potential benefits that promote healthy aging. Here, we use Caenorhabditis elegans as a model to show that water-soluble cranberry extract standardized to 4.0% proanthocyanidins (WCESP), a major component of cranberries, can enhance host innate immunity to resist against Vibrio cholerae (V. cholerae; wild type C6706 (O1 El Tor biotype)) infection. Supplementation of WCESP did not significantly alter the intestinal colonization of V. cholerae, but upregulated the expression of C. elegans innate immune genes, such as clec-46, clec-71, fmo-2, pqn-5 and C23G10.1. Additionally, WCESP treatment did not affect the growth of V. cholerae and expression of the major bacterial virulence genes, and only slightly reduced bacterial colonization within C. elegans intestine. These findings indicate that the major components of WCESP, including proanthocyanidins (PACs), may play an important role in enhancing the host innate immunity. Moreover, we engaged C. elegans mutants and identified that the p38 MAPK signaling, insulin/IGF-1 signaling (IIS), and HSF-1 play pivotal roles in the WCESP-mediated host immune response. Considering the level of conservation between the innate immune pathways of C. elegans and humans, the results of this study suggest that WCESP may also play an immunity-promoting role in higher order organisms.
Assuntos
Proteínas de Caenorhabditis elegans/biossíntese , Imunidade Inata/efeitos dos fármacos , Extratos Vegetais/administração & dosagem , Proantocianidinas/administração & dosagem , Fatores de Transcrição/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Envelhecimento/efeitos dos fármacos , Envelhecimento/genética , Animais , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/genética , Caenorhabditis elegans/imunologia , Proteínas de Caenorhabditis elegans/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Fator de Crescimento Insulin-Like I/biossíntese , Extratos Vegetais/química , Proantocianidinas/química , Transdução de Sinais/efeitos dos fármacos , Fatores de Transcrição/genética , Vaccinium macrocarpon/química , Vibrio cholerae/efeitos dos fármacos , Vibrio cholerae/patogenicidade , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
PKCη is involved in proliferation, differentiation, and drug resistance. However, PKCη function in EBV(+) B lymphoma remains poorly understood. Gene silencing of PKCη through siRNA knockdown inhibited cellular proliferation, induced cell cycle arrest in G0/G1 and G2/M phases, and sensitized cells to chemotherapeutic drugs. Upon PKCη knockdown, expression levels of p21, GADD45α, and TAp73 were all increased, whereas expression levels of CDK2, CDK4, CDK6, cyclin E, cyclin B1, and cdc2 were all downregulated. PKCη silencing also activated p38-MAPK, which in turn contributed to the expression of cell cycle arrest-related molecules. These results suggest that siRNA-mediated silencing of PKCη can be a potent tool to complement existing chemotherapy regimens for treating EBV(+) B lymphoma.
Assuntos
Antineoplásicos/uso terapêutico , Linfoma de Burkitt/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos/genética , Herpesvirus Humano 4 , Proteína Quinase C/genética , Apoptose/efeitos dos fármacos , Apoptose/genética , Ácidos Borônicos/uso terapêutico , Bortezomib , Linfoma de Burkitt/genética , Pontos de Checagem do Ciclo Celular/genética , Proteínas de Ciclo Celular/biossíntese , Divisão Celular/genética , Linhagem Celular Tumoral , Proliferação de Células , Inibidor de Quinase Dependente de Ciclina p21/biossíntese , Inibidor de Quinase Dependente de Ciclina p21/genética , Proteínas de Ligação a DNA/biossíntese , Humanos , Potencial da Membrana Mitocondrial , NF-kappa B/genética , Niacinamida/análogos & derivados , Niacinamida/uso terapêutico , Proteínas Nucleares/biossíntese , Compostos de Fenilureia/uso terapêutico , Fosfatidilinositol 3-Quinases/genética , Proteína Quinase C/metabolismo , Pirazinas/uso terapêutico , Interferência de RNA , RNA Interferente Pequeno/genética , Sorafenibe , Proteína Tumoral p73 , Proteínas Supressoras de Tumor/biossíntese , Regulação para Cima , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
Paeoniflorin (PF) is one of the principal components of peony, a plant widely used in traditional Chinese medicine for its anti-inflammatory and immunomodulatory effects. Human ß-defensin-2 (hBD-2) is an antimicrobial peptide that acts as the first line of defense against bacterial, viral, and fungal infections. This study aims to determine whether or not PF can regulate the expression of hBD-2 and its possible molecular mechanism in human bronchial epithelial cells (HBECs). Real-time quantitative reverse transcription PCR showed that PF can enhance the mRNA expression level of hBD-2 in a concentration- and time-dependent manner in HBECs. Further studies demonstrated that the mRNA and protein expression levels of hBD-2 were attenuated by the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580, the extracellular signal-regulated kinase (ERK) inhibitor PD98059, and the nuclear factor kappa B (NF-κB) inhibitor (pyrrolidine dithiocarbamate (PDTC)). The phosphorylation of p38 MAPK, ERK, and c-Jun N-terminal kinase was detected by Western blot analysis, and the NF-κB translocation of 16HBECs after PF treatment was analyzed by immunofluorescence. These results support that PF upregulates hBD-2 expression in HBECs through the p38 MAPK, ERK, and NF-κB signaling pathways. These findings provide a new pharmacological mechanism of PF for the treatment of microbial infections by strengthening epithelial antimicrobial barriers.
Assuntos
Glucosídeos/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Monoterpenos/farmacologia , NF-kappa B/biossíntese , Mucosa Respiratória/metabolismo , beta-Defensinas/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Linhagem Celular Transformada , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica , Humanos , Sistema de Sinalização das MAP Quinases/fisiologia , Paeonia , Mucosa Respiratória/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/fisiologiaRESUMO
AIMS AND BACKGROUND: Ginsenoside Rh2, which exerts the potent anticancer action both in vitro and in vivo, is one of the most well characterized ginsenosides extracted from ginseng. Although its effects on cancer are significant, the underlying mechanisms remain unknown. In this study, we sought to elucidate possible links between ginsenoside Rh2 and phosphoglucose isomerase/autocrine motility factor (PGI/AMF). METHODS: KG1α, a leukemia cell line highly expressing PGI/AMF was assessed by western blot analysis and reverse transcription- PCR (RT-PCR) assay after transfection of a small interfering (si)-RNA to silence PGI/AMF. The effect of PGI/ AMF on proliferation was measured by typan blue assay and antibody array. A cell counting kit (CCK)-8 and flow cytometry (FCM) were adopted to investigate the effects of Rh2 on PGI/AMF. The relationships between PGI/AMF and Rh2 associated with Akt, mTOR, Raptor, Rag were detected by western blot analysis. RESULTS: KG1α cells expressed PGI/AMF and its down-regulation significantly inhibited proliferation. The antibody array indicated that the probable mechanism was reduced expression of PARP, State1, SAPK/JNK and Erk1/2, while those of PRAS40 and p38 were up-regulated. Silencing of PGI/AMF enhanced the sensibility of KG1α to Rh2 by suppressing the expression of mTOR, Raptor and Akt. CONCLUSION: These results suggested that ginsenoside Rh2 suppressed the proliferation of KG1α, the same as down-regulation of PGI/AMF. Down-regulation of PGI/ AMF enhanced the pharmacological effects of ginsenoside Rh2 on KG1α by reducing Akt/mTOR signaling.
Assuntos
Glucose-6-Fosfato Isomerase/genética , Leucemia/tratamento farmacológico , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Panax/metabolismo , Extratos Vegetais/farmacologia , Proteínas Adaptadoras de Transdução de Sinal/biossíntese , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células , Regulação para Baixo , MAP Quinases Reguladas por Sinal Extracelular/biossíntese , Expressão Gênica/efeitos dos fármacos , Células HL-60 , Proteínas de Homeodomínio/biossíntese , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/biossíntese , Poli(ADP-Ribose) Polimerases/biossíntese , Proteínas Proto-Oncogênicas c-akt , Interferência de RNA , RNA Interferente Pequeno , Proteína Regulatória Associada a mTOR , Fator de Transcrição STAT1/biossíntese , Serina-Treonina Quinases TOR/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
Liver cancer is a malignant tumour with high morbidity and fatality rates that is common worldwide. At present, the clinical approaches to treating primary liver cancer include partial hepatectomy, systemic or local chemotherapy, radiotherapy, radiofrequency ablative surgery and liver transplantation. However, all of these approaches have shortcomings, including poor prognosis and numerous side-effects. A large number of studies have proven that many effective ingredients in traditional Chinese medicine, particularly the flavonoid compounds extracted from plants, have achieved breakthroughs in terms of enhancing the effects and reducing the toxicity of chemotherapy and radiotherapy, preventing tumour metastasis and relapse after surgery, alleviating the clinical symptoms of advanced tumours, improving the quality of life of the patient with tumours and extending patient longterm survival. The purpose of the present study was to investigate the impact of isoquercitrin, the flavonoid from Bidens bipinnata L. extract, on the progression of liver cancer and to achieve a deeper understanding of the biological characteristics of isoquercitrin's involvement in the progression of liver cancer. In the in vitro experiments, isoquercitrin was found to strongly inhibit the proliferation of human liver cancer cells, promote the apoptosis of human liver cancer cells, and block the cell cycle in the G1 phase. Isoquercitrin activated caspase-3, -8 and -9, inhibited the expression level of ERK and p38MAPK protein phosphorylation, and promoted the phosphorylation of JNK. Additionally, isoquercitrin reduced the expression level of PKC in human liver cancer cells. In the in vivo experiments, isoquercitrin was also found to significantly inhibit the growth of transplanted tumours in nude mice. The present study confirmed that isoquercitrin could inhibit the progression of human liver cancer in vivo and in vitro, and the molecular mechanism of isoquercitrin may be closely associated with the MAPK and PKC signalling pathways.
Assuntos
Neoplasias Hepáticas/tratamento farmacológico , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Quercetina/análogos & derivados , Animais , Apoptose/efeitos dos fármacos , Caspase 3/biossíntese , Caspase 8/biossíntese , Caspase 9/biossíntese , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Progressão da Doença , MAP Quinases Reguladas por Sinal Extracelular/biossíntese , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Células Hep G2 , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Neoplasias Hepáticas/prevenção & controle , Masculino , Medicina Tradicional Chinesa , Camundongos , Camundongos Nus , Fosforilação , Quercetina/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
This study investigated the effects of purple sweet potato leaf extract (PSPLE) and its components, cyanidin and quercetin, on human aortic endothelial cells (HAECs) during the inflammatory process. HAECs were pretreated with 100 µg/mL PSPLE or 10 µM quercetin, cyanidin or aspirin for 18 h followed by TNF-α (2 ng/mL) for 6 h, and U937 cell adhesion was determined. Adhesion molecule expression and CD40 were evaluated; NFκB p65 protein localization and DNA binding were assessed. PSPLE, aspirin, cyanidin and quercetin significantly inhibited TNF-α-induced monocyte-endothelial cell adhesion (p < 0.05). Cyanidin, quercetin and PSPLE also significantly attenuated VCAM-1, IL-8 and CD40 expression, and quercetin significantly attenuated ICAM-1 and E-selectin expression (p < 0.05). Significant reductions in NFκB expression and DNA binding by aspirin, cyanidin and quercetin were also observed in addition to decreased expression of ERK1, ERK2 and p38 MAPK (p < 0.05). Thus, PSPLE and its components, cyanidin and quercetin, have anti-inflammatory effects through modulation of NFκB and MAPK signaling. Further in vivo studies are necessary to explore the possible therapeutic effects of PSPLE on atherosclerosis.
Assuntos
Adesão Celular/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Inflamação/tratamento farmacológico , Ipomoea batatas/química , Extratos Vegetais/farmacologia , Antocianinas/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Aorta/efeitos dos fármacos , Aspirina/farmacologia , Antígenos CD40/biossíntese , Moléculas de Adesão Celular/biossíntese , Linhagem Celular , Selectina E/biossíntese , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Humanos , Molécula 1 de Adesão Intercelular/biossíntese , Interleucina-8/biossíntese , Proteína Quinase 1 Ativada por Mitógeno/biossíntese , Proteína Quinase 3 Ativada por Mitógeno/biossíntese , NF-kappa B/biossíntese , Folhas de Planta/química , Quercetina/farmacologia , Fator de Transcrição RelA/efeitos dos fármacos , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Células U937 , Molécula 1 de Adesão de Célula Vascular/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
Gliomas are a group of heterogeneous primary central nervous system tumors. Hyperthermia has proven to be a potential therapeutic tool for cancers in the clinic. However, the molecular mechanisms of hyperthermia remain unclear. The objective of this study was to investigate the effects of hyperthermia on the invasiveness in C6 glioma cells and related molecular pathways. Here our data show hyperthermia stimulated the release of tumor necrosis factor-alpha (TNF-α) and decreased C6 glioma cell migration and invasive capability at 30, 60, 120 and 180 min; with increased spontaneous apoptosis in C6 glioma cells at 120 min. We also found mitogen-activated protein kinase (P38 MAPK) protein expression to be increased and nuclear factor-kappa B (NF-κB) protein expression decreased. Based on the results, we conclude that hyperthermia alone reduced invasion of C6 glioma cells through stimulating TNF-α signaling to activate apoptosis, enhancing P38 MAPK expression and inhibiting the NF-κB pathway, a first report in C6 rat glioma cells.
Assuntos
Apoptose/fisiologia , Neoplasias Encefálicas/terapia , Glioma/patologia , Glioma/terapia , Hipertermia Induzida/métodos , Animais , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Hipóxia Celular/fisiologia , Linhagem Celular Tumoral , Movimento Celular/genética , Glioma/genética , Glioma/metabolismo , NF-kappa B/biossíntese , NF-kappa B/genética , Invasividade Neoplásica , Ratos , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genética , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Helonias dioica (HD) is a threatened species of herb growing in North America. It is used as a traditional medicine for treating various ailments particularly related to reproductive issues. The root is reported to contain approximately 10% of a saponin (chamaelirin; C(36)H(62)O(18)) apart from certain other fatty acids. As saponins are known to have hypoglycemic effects, we suspected its possible antihyperglycemic potentials. We injected intraperitoneally alloxan (ALX) at the dose of 200 mg/kg body weight (bw) to induce hyperglycemia in mice and tested possible hypoglycemic effects of HD in vivo by deploying two doses (100 and 200 mg/kg bw, respectively). We also tested its effects on the isolated pancreatic islets cells in vitro. We used various standard protocols like reactive oxygen species (ROS) generation and DNA damage, activities of biomarkers like catalase (CAT), superoxide dismutase (SOD), lipid peroxidase (LPO), reduced glutathione (GSH) of the pancreas tissue and glucokinase and glycogen content of the liver of hyperglycemic mice. With a mechanistic approach, we also tracked down the possible signaling pathway involved. We found an elevated level of ROS generation, LPO and overexpression of inducible nitric oxide synthase (iNOS), tumor necrosis factor α (TNF-α), p38 Map kinase (p38 MAPK), nuclear factor (NF)-κß, interferon gamma (IFN-γ), cytochrome c, caspase 3, poly [ADP ribose] polymerase (PARP) and cyclo oxygenase 2 (COX2) in ALX-induced diabetic mouse. Treatment of hyperglycemic mice with both the doses of HD showed a significant decrease with respect to all these parameters of study. Thus, our results suggest that HD prevents ALX-induced islet cell damage and possesses antihyperglycemic and antioxidative potentials.
Assuntos
Hiperglicemia/tratamento farmacológico , Hipoglicemiantes/farmacologia , Magnoliopsida , Estresse Oxidativo/efeitos dos fármacos , Fitoterapia , Extratos Vegetais/farmacologia , Plantas Medicinais/metabolismo , Aloxano , Animais , Caspase 3/biossíntese , Catalase/metabolismo , Células Cultivadas , Ciclo-Oxigenase 2/biossíntese , Citocromos c/biossíntese , Dano ao DNA , Espécies em Perigo de Extinção , Glucoquinase/metabolismo , Glutationa/metabolismo , Hiperglicemia/induzido quimicamente , Hipoglicemiantes/administração & dosagem , Ilhotas Pancreáticas/efeitos dos fármacos , Peróxidos Lipídicos/metabolismo , Glicogênio Hepático/análise , Magnoliopsida/química , Camundongos , NF-kappa B/biossíntese , Óxido Nítrico Sintase Tipo II/biossíntese , Pâncreas/metabolismo , Extratos Vegetais/administração & dosagem , Raízes de Plantas/química , Poli(ADP-Ribose) Polimerases/biossíntese , Espécies Reativas de Oxigênio/metabolismo , Saponinas/análise , Saponinas/farmacologia , Transdução de Sinais , Superóxido Dismutase/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
Oxidative stress significantly contributes to cisplatin (CP)-associated cytotoxicity, and use of antioxidants could counteract such cytotoxic effects of CP. The major biochemical pathway for reactive oxygen species (ROS) formation proceeds through O2â» production, which is generated by NADPH oxidase, such oxidative stress can activate p38 MAPK to intensify the cytotoxic effect of CP. We mainly aimed to study the protective effect of oxytocin (OT) on CP-induced nephrotoxicity whereas; it was previously shown to have anti-inflammatory effects in different inflammation models. Administration of OT significantly decreased the gene expression of both NADPH oxidase and P38 MAPK, nitric oxide (NO), myloperoxidase (MPO), and TBARS, furthermore it increased the renal tissue levels of antioxidants; reduced glutathione (GSH), and superoxide dismutase (SOD). Histologically, OT reduced the monocellular infiltration as well as the tubular damage in CP-induced nephrotoxicity. In conclusion OT has a powerful antioxidant effect that can alleviate the CP-induced nephrotoxicity through inhibition of NADPH oxidase and P38 MAPK resulting in improvement of kidney functions.
Assuntos
Cisplatino/toxicidade , Nefropatias/induzido quimicamente , NADPH Oxidases/antagonistas & inibidores , Ocitocina/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Avaliação Pré-Clínica de Medicamentos , Indução Enzimática/efeitos dos fármacos , Córtex Renal/efeitos dos fármacos , Córtex Renal/patologia , Nefropatias/enzimologia , Nefropatias/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , NADPH Oxidases/biossíntese , NADPH Oxidases/genética , Óxido Nítrico/biossíntese , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Peroxidase/biossíntese , Peroxidase/genética , Distribuição Aleatória , Ratos , Ratos Wistar , Superóxido Dismutase/biossíntese , Superóxido Dismutase/genética , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Proteínas Quinases p38 Ativadas por Mitógeno/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Cordyceps species have been known since long as a multi-utility ethnomedicinal herbal in Korea, China and Japan. It has been reported to exhibit a number of properties such as anti-oxidative, anti-cancer, antiinflammatory, anti-diabetic, and anti-obesity effects. In a previously conducted study, we had demonstrated that the ethanol extract of Cordyceps bassiana was able to suppress the production of interleukin (IL)-12 and interferon (IFN)-gamma in macrophages and T lymphocytes. In this study, we were able to further explore the molecular basis of its inhibitory mechanism using a butanol fraction of this herbal (Cb-BF) preparation. Similarly, this fraction also blocked the expression of cytokines such as IL-12 and tumor necrosis factor (TNF)-alpha as well as the proliferation of splenic lymphocytes and their production of IFN-gamma but not IL-4. Cb-BF suppressed the luciferase activities that are mediated by nuclear factor (NF)-kappaB, activator protein (AP)-1, and signal transducers and activators of transcription (STAT)-1. In agreement with this, these fractions diminished the translocation of the transcription factors into the nucleus. The study also demonstrated that the upstream signaling events for the activation of these factors such as spleen tyrosine kinase (Syk), janus kinase (JAK)-2, and extracellular signal-regulated kinase (ERK) were suppressed. Therefore, these results suggest that the butanol extract of Cordyceps bassiana may contain more than one active component capable of inhibiting the inflammatory signaling cascade and this can be considered as a potential candidate for treatment of diseases that require suppression of immune system.
Assuntos
Cordyceps/química , Citocinas/antagonistas & inibidores , Citocinas/biossíntese , Animais , Western Blotting , Butanóis , Corantes , Genes Reporter/efeitos dos fármacos , Humanos , Interferon gama/biossíntese , Interleucina-12/biossíntese , Interleucina-4/biossíntese , Lipopolissacarídeos/farmacologia , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , Extratos Vegetais/farmacologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Transdução de Sinais/efeitos dos fármacos , Solventes , Baço/citologia , Baço/efeitos dos fármacos , Sais de Tetrazólio , Tiazóis , Fatores de Transcrição , Fator de Necrose Tumoral alfa/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/biossínteseRESUMO
Dendrobium alkaloids (DNLA), extracted from Dendrobium nobile Lindl. whose botanical name is Dendrobium moniliforme, Orchidaceae family, were studied for their effect on lipopolysaccharide (LPS)-induced memory impairment in rats. SD rats were pretreated with DNLA (40, 80, 160 mg/kg/d for 7 d), followed by LPS (50 µg) injection into the right lateral ventricle to produce memory impairment. DNLA treatment continued for another 13 days. The spatial behavior was tested by the Morris water maze; the level of tumor necrosis factor receptor 1 (TNFR1) mRNA was detected by real time RT-PCR, and the protein level of TNFR1, nuclear factor kappa-B (NF- κB) and phosphorylated p38 mitogen-activated protein kinases (p-p38 MAPK) by Western blotting. The results showed that DNLA significantly improved the neurobehavioral performance and prevented LPS-induced elevation in TNFR1 mRNA and protein levels. LPS-induced activation of p38 MAPK and NF- κB pathway was also suppressed. In conclusion, DNLA is effective in protecting against LPS-induced brain impairment, and this effect is due, at least in part, to prevent overexpression of TNFR1 via inhibition of p-p38 MAPK and the downstream NF- κB signal pathway.