RESUMO
Two new chromones named cnidimol G (1) and cnidimol H (2), one new coumarin, 7-methoxy-8-(3-methoxy-3-methyl-2-oxobutyl)coumarin (3), and twenty known compounds were isolated from MeOH extract of the fruit of Cnidium monnieri (L.) Cusson. The structures of compounds were elucidated by extensive spectroscopic analyses including 1 D and 2 D NMR, HRESIMS, IR and UV. Anti-inflammatory activity of the selected isolated compounds were evaluated. Compounds 1 and 8 exhibited inhibitory activities against nitric oxide production.
Assuntos
Cnidium , Frutas , Cnidium/química , Frutas/química , Cromonas/farmacologia , Cromonas/análise , Extratos Vegetais/química , Cumarínicos/químicaRESUMO
Purine compounds are known to activate 5'-adenosine monophosphate-activated protein kinase (AMPK), which has important roles in treatments for renal cell carcinoma. The present study was aimed to investigate the effects of the purine analogue ENERGIF706 on the human renal carcinoma cell line 786O and the underlying mechanisms. The results revealed that ENERGIF706 (0.20.6 mg/ml) significantly decreased the cell viability to up to 36.4±2.4% of that of the control. Compared to 786O cells, ENERGIF706 exerted less suppressive effects on the viability of the human nontumorigenic renal cell line HK2. Flow cytometric analysis showed that ENERGIF706 contributed to cell cycle arrest at Sphase and triggered apoptosis of 786O cells. Immunoblot analysis revealed that antiapoptotic Bcell lymphoma 2 (Bcl2) levels were reduced and proapoptotic Bcl2associated X protein levels were diminished. In addition, activation of caspase9, caspase3 and poly(adenosine diphosphate ribose) polymerase (PARP) was promoted in 786O cells in response to ENERGIF706. Effects of ENERGIF706 on AMPK cascades were investigated and the results showed that ENERGIF706 enhanced phosphorylation of AMPKα (T172) and p53 (S15), a downstream target of AMPK. In addition, the AMPK activation, p53 (S15) phosphorylation, reduction of Bcl2, cleavage of caspase3 and PARP as well as suppressed cell viability induced by ENERGIF706 were reversed in the presence of AMPK inhibitor compound C (dorsomorphin). In conclusion, the findings of the present study revealed that ENERGIF706 significantly suppressed the viability of 786O cells via induction of cell cycle arrest and apoptosis, attributing to AMPK and p53 activation and subsequent cell cycle regulatory and apoptotic signaling. It was therefore indicated that ENERGIF706 may be suitable for the treatment of renal cell carcinoma.
Assuntos
Proteínas Quinases Ativadas por AMP/genética , Antineoplásicos Fitogênicos/farmacologia , Bambusa/química , Células Epiteliais/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica , Purinas/farmacologia , Proteínas Quinases Ativadas por AMP/metabolismo , Antineoplásicos Fitogênicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Ativação Enzimática/efeitos dos fármacos , Células Epiteliais/enzimologia , Células Epiteliais/patologia , Humanos , Rim/efeitos dos fármacos , Rim/enzimologia , Rim/patologia , Especificidade de Órgãos , Extratos Vegetais/química , Poli(ADP-Ribose) Polimerases/genética , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Purinas/isolamento & purificação , Pirazóis/farmacologia , Pirimidinas/farmacologia , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Transdução de Sinais , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
Adenosine monophosphate (AMP)-activated protein kinase (AMPK) plays a central role in energy homeostasis and regulation of inflammatory responses. The present study is aimed to investigate the anti-inflammatory effects of ENERGI-F704, a nucleobase analogue isolated from bamboo leaves, on expression of proinflammatory mediators in murine macrophage RAW264.7 in response to lipopolysaccharide (LPS). ENERGI-F704 enhanced phosphorylation of AMPK(T172) but insignificantly affected the viability of RAW264.7 cells. Further investigation showed that ENERGI-F704 decreased mRNA expression of interleukin (IL)-6, IL-8, tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX2), and inducible nitric oxide synthase (iNOS) induced by LPS, as well as suppressed the production of prostaglandin E2 (PGE2) and nitric oxide (NO). Additionally, the inhibitory effects of ENERGI-F704 on the LPS-induced proinflammatory mediators were diminished by pretreatment of AMPK inhibitor Compound C. ENERGI-F704 also inhibited LPS-triggered activation of nuclear factor kappa B (NF-κB), phosphatidylinositol 3-kinase (PI3K), and p38 mitogen-activated protein kinase (p38), whereas extracellular signal-regulated kinase (Erk)1/2 and c-Jun N-terminal kinase (JNK) were insignificantly influenced. Our findings indicate that ENERGI-F704 may exert anti-inflammatory activity on RAW264.7 cells in response to LPS through the activation of AMPK and suppression of PI3K/P38/NF-κB signaling and the consequent decreased expression of proinflammatory mediators, suggesting that ENERGI-F704 is beneficial to the amelioration of inflammatory disorders.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Mediadores da Inflamação/metabolismo , Macrófagos/metabolismo , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/genética , Citocinas/metabolismo , Regulação para Baixo/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Immunoblotting , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Camundongos , Fosforilação/efeitos dos fármacos , Extratos Vegetais/farmacologia , Folhas de Planta/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sasa/química , Transdução de Sinais/efeitos dos fármacosRESUMO
Perilla leaves are widely used in Chinese herbal medicine and in Japanese herbal agents used to treat respiratory diseases. This study aimed to investigate the antiinflammatory effects and the underlying mechanisms of Perilla frutescens leaf extract (PLE). Murine macrophage RAW264.7 cells were used as a model. Cell viability and morphological changes were studied by the MTT assay and microscopy. mRNA expression of proinflammatory mediators was assessed by both semiquantitative reverse transcriptionpolymerase chain reaction (RTPCR) and quantitative (q) RTPCR. Nitric oxide (NO) and prostaglandin E2 (PGE2) production were analyzed by the Griess test and sandwich enzymelinked immunosorbent assay (ELISA), respectively. The activation of kinase cascades was studied by immunoblotting. Our findings showed that PLE slightly affects cell viability, but alleviates LPSinduced activation of RAW264.7 cells. Furthermore, PLE significantly reduced the LPSinduced mRNA expression of the interleukin (IL)6, IL8, tumor necrosis factorα (TNFα), cyclooxygenase2 (COX2) and inducible nitric oxide synthase (iNOS), genes in a dosedependent manner. In addition, PLE reduced NO production and PGE2 secretion induced by LPS. PLE also inhibited activation of mitogenactivated protein kinases (MAPKs), increased the cytosolic IκBα level, and reduced the level of nuclear factor (NF)κB. Taken together, these findings indicate that PLE significantly decreases the mRNA expression and protein production of proinflammatory mediators, via the inhibition of extracellularsignalregulated kinase (ERK)1/2, cJun Nterminal kinase (JNK), p38, as well as NFκB signaling in RAW264.7 cells stimulated with LPS.
Assuntos
Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Perilla frutescens/química , Extratos Vegetais/farmacologia , Animais , Anti-Inflamatórios/química , Linhagem Celular , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Expressão Gênica/efeitos dos fármacos , Proteínas I-kappa B/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Lipopolissacarídeos/toxicidade , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Inibidor de NF-kappaB alfa , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Perilla frutescens/metabolismo , Fosforilação/efeitos dos fármacos , Extratos Vegetais/química , Folhas de Planta/química , Folhas de Planta/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Perilla frutescens has been used in traditional medicine for respiratory diseases due to its anti-bacterial and anti-inflammatory activity. This study aimed to investigate effects of Perilla frutescens leaf extract (PFE) on expression of pro-allergic and pro-inflammatory cytokines in airway epithelial cells exposed to mite major allergen Der p 2 (DP2) and the underlying mechanisms. Our results showed that PFE up to 100 µg/mL had no cytotoxic effect on human bronchial epithelial cell BEAS-2B. Further investigations revealed that PFE dose-dependently diminished mRNA expression of pro-allergic cytokine IL-4, IL-5, IL-13 and GM-CSF, as well as pro-inflammatory cytokine IL-6, IL-8 and MCP-1 in BEAS-2B cells treated with DP2. In parallel to mRNA, the DP-2-elevated levels of the tested cytokines were decreased. Further investigation showed that DP2-indued phosphorylation of p38 MAPK (P38) and JNK, but not Erk1/2, was also suppressed by PFE. In addition, PFE elevated cytosolic IκBα level and decreased nuclear NF-κB level in DP2-stimulated BEAS-2B cells. Taken together, these findings revealed that PFE significantly diminished both mRNA expression and protein levels of pro-allergic and pro-inflammatory cytokines in response to DP2 through inhibition of P38/JNK and NK-κB activation. These findings suggest that PFE should be beneficial to alleviate both allergic and inflammatory responses on airway epithelium in response to aeroallergens.