RESUMO
Triple-Negative Breast Cancer (TNBC) is a most dangerous breast cancer subtype. The naturally occurring sesquiterpene lactone, arnicolide D (AD), has proven effective against a variety of tumors, however, the inhibitory effects of AD against TNBC and the underlying mechanisms remain unclear. In the present study, two TNBC cell lines (MDA-MB-231 and MDA-MB-468) and an MDA-MB-231 xenograft mouse model were employed to investigate the anti-TNBC effects of AD in vitro and in vivo. Cell viability was assessed by MTT assay. Cell cycle arrest and apoptosis were analyzed by flow cytometry. Protein levels were determined by immunoblotting. In vitro studies demonstrated that AD significantly decreased cell viability, and induced G2/M cell cycle arrest and apoptosis. In vivo assays showed that oral administration of 25 or 50 mg/kg AD for 22 days led to a reduction of tumor weights by 24.7% or 41.0%, without appreciable side effects. Mechanistically, AD inhibited the activation of Akt/mTOR and STAT3 signaling pathways. Based on our findings, AD is a promising candidate for development as an adjunctive therapeutic drug for TNBC.
Assuntos
Lactonas/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Transcrição STAT3/metabolismo , Sesquiterpenos/uso terapêutico , Serina-Treonina Quinases TOR/metabolismo , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/metabolismo , Animais , Western Blotting , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Lactonas/farmacologia , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas Proto-Oncogênicas c-akt/genética , Fator de Transcrição STAT3/genética , Sesquiterpenos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Accumulating evidence suggests that hepatic stellate cells (HSCs) adopt aerobic glycolysis during activation. Hedgehog (Hh) pathway plays a vital role in the process of HSCs activation by regulating metabolism, and activation of the Hh pathway promotes transdifferentiation of HSCs into myofibroblasts. Deoxyelephantopin (DET), a naturally occurring sesquiterpene lactone from Elephantopus scaber, has been shown to exert hepatoprotective as well as anticancer effects. However, the effect of DET on hepatic fibrosis and glycolysis in HSCs have never been elucidated. Here, we studied the function of the DET on HSCs activation and investigated the anti-fibrogenic effects of DET was associated with interfering with glycolysis in HSCs. Our results first demonstrated that DET reduced the expression of a-smooth muscle actin (a-SMA) and a1(I)procollagen at both mRNA and protein levels, and restore lipogenesis in HSCs. Furthermore, DET decreased the expression of hexokinase (HK), phosphofructokinase-2 (PFK2), Glucose transporter 4 (Glut4), and reduced lactate production dose-dependently in HSCs. Moreover, we further revealed that DET reduced fibrotic gene expression, restored lipid accumulation in HSCs. However, the Hh pathway agonist SAG could reverse the above effect of DET. Together, these results indicate DET inhibits aerobic glycolysis in HSCs associated with inhibition of Hh pathway. Our results provided a novel mechanism for DET suppression of HSC activation implicated in antifibrotic therapy.
Assuntos
Proteínas Hedgehog/metabolismo , Células Estreladas do Fígado/efeitos dos fármacos , Lactonas/farmacologia , Substâncias Protetoras/farmacologia , Sesquiterpenos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Asteraceae/química , Linhagem Celular , Glicólise/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Lactonas/química , Lipogênese/efeitos dos fármacos , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/metabolismo , Substâncias Protetoras/química , Ratos , Sesquiterpenos/químicaRESUMO
Curculiginis Rhizoma (Curculigo orchioides GAERTN.) is a well-known Chinese herbal medicine, as well as an important Rasayana drug in India. Current criteria of quality control on this herb are to quantitatively analyze single compound curculigoside, which fail to comprehensively evaluate quality of this herb. In this paper, a simple and reliable HPLC coupled with diode array detector (DAD) method was developed to evaluate the quality of Curculiginis Rhizoma through establishing chromatographic fingerprint and simultaneously quantitating four phenolic compounds, orcinol glucoside, orcinol, 2,6-dimethoxybenzoic acid and curculigoside. The fingerprint displayed eleven common peaks, and the similarity index of different samples was in a range of 0.890-0.977. Validation of the method was acceptable, with 96.03-102.82% accuracy in recovery test and inter and intra-day precisions were less than 2%. This developed method by having a combination of chromatographic fingerprint and quantitation analysis could be applied to the quality control of Curculiginis Rhizoma.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Curculigo/química , Medicamentos de Ervas Chinesas/química , Fenóis/análise , Rizoma/química , Calibragem , Glucosídeos/análise , Éteres de Hidroxibenzoatos/análise , Controle de QualidadeRESUMO
Two new diarylheptanoids, 1-(3',5'-dihydroxy-4'-methoxyphenyl)-7-phenyl-3-heptanone (1) and 1-(2',4'-dihydroxy-3'-methoxyphenyl)-7-(4â³-methoxyphenyl)-3-heptanone (2), along with known diarylheptanoid yakuchinone A (3), and five flavanoids, tectochrysin (4), chrysin (5), izalpinin (6), kaempferol 7, 4'-dimethyl ether (7), and kaempferide (8) were isolated from the fruits of Alpinia oxyphylla Miq. Their structures were determined by means of spectroscopic methods. Antioxidant activities of all the isolated compounds were evaluated using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Compounds 1-3 and 6-8 exhibited potent antioxidant activities in the DPPH assay.
Assuntos
Alpinia/química , Antioxidantes/isolamento & purificação , Diarileptanoides/isolamento & purificação , Diarileptanoides/farmacologia , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Antioxidantes/química , Antioxidantes/farmacologia , Compostos de Bifenilo/farmacologia , Diarileptanoides/química , Medicamentos de Ervas Chinesas/química , Flavonoides/química , Frutas/química , Guaiacol/análogos & derivados , Guaiacol/química , Guaiacol/isolamento & purificação , Quempferóis/isolamento & purificação , Quempferóis/farmacologia , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Picratos/farmacologiaRESUMO
PURPOSE: Triple-negative breast cancer (TNBC) is a a breast cancer subtype characterized by a lack of estrogen receptor, progesterone receptor and human epidermal growth receptor 2 and is associated with poorer prognoses when compared to other breast cancers. Thus, novel anti-cancer agents with high efficacy are urgently needed. Brevilin A (BA), a natural sesquiterpene lactone, has been reported to exhibit anti-cancer effects. However, the effects of BA on TNBC have not yet been demonstrated. In this study, we investigated the anti-TNBC effects and the underlying mechanism of BA, in vitro and in vivo. METHODS: Two TNBC cell lines and a xenograft mouse model were employed to assess the effects of BA. Cell viability was detected by MTT assay. Cell cycle status and apoptosis were evaluated by flow cytometry. Cell migration was measured by wound-healing assay. Protein expression was measured by Western blotting analysis. The in vivo anti-cancer activity of BA was assessed in orthotopic tumor xenograft mice. RESULTS: BA significantly inhibited the growth of TNBC cells in a dose- and time-dependent manner via induction of cell cycle arrest at G2/M phase arrest and apoptosis. BA also inhibited tumor cell migration. BA significantly downregulated the expression of Akt, mTOR, Stat3 and their phosphorylation, and thus inhibiting the activation of the Akt/mTOR and STAT3 signaling pathways. Furthermore, oral administration of BA at 25 or 50 mg/kg leads to significant inhibition of tumor growth and proliferation in tumor xenograft model mice. CONCLUSION: BA significantly inhibited the growth and migration of TNBC cells, and induced cell cycle arrest and apoptosis. These inhibitory effects were associated with the suppression of the Akt/mTOR and Stat3 signal pathways. Based on our findings, BA possesses a promising candidate for development as an anti-cancer therapeutic drug against TNBC.
RESUMO
Growing evidence suggests that activated immune cells undergo metabolic reprogramming in the regulation of the innate inflammatory response. Remarkably, macrophages activated by lipopolysaccharide (LPS) induce a switch from oxidative phosphorylation to aerobic glycolysis, and consequently results in release of proinflammatory cytokines. Pyruvate Kinase M2 (PKM2) plays a vital role in the process of macrophage activation, promoting the inflammatory response in sepsis and septic shock. Deoxyelephantopin (DET), a naturally occurring sesquiterpene lactone from Elephantopus scaber, has been shown to counteracts inflammation during fulminant hepatitis progression, but the underlying mechanism remains unclear. Here, we studied the function of the DET on macrophage activation and investigated the anti-inflammatory effects of DET associated with interfering with glycolysis in macrophage. Our results first demonstrated that DET attenuates LPS-induced interleukin-1ß (IL-1ß) and high-mobility group box 1 (HMGB1) release in vitro and in vivo and protected mice against lethal endotoxemia. Furthermore, DET decreased the expression of pyruvate dehydrogenase kinase 1 (PDK1), glucose transporter 1(GLUT1), lactate dehydrogenase A (LDHA), and reduced lactate production dose-dependently in macrophages. Moreover, we further revealed that DET attenuates aerobic glycolysis in macrophages associated with regulating the nuclear localization of PKM2. Our results provided a novel mechanism for DET suppression of macrophages activation implicated in anti-inflammatory therapy.
Assuntos
Anti-Inflamatórios/uso terapêutico , Lactonas/uso terapêutico , Macrófagos/imunologia , Piruvato Quinase/metabolismo , Sepse/tratamento farmacológico , Sesquiterpenos/uso terapêutico , Aerobiose , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Glicólise/efeitos dos fármacos , Humanos , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/imunologia , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células RAW 264.7 , Sepse/imunologia , Transdução de SinaisRESUMO
OBJECTIVE: To develop a GC method to determine the content of caryophyllene in Eupatorium fortune. METHOD: The samples were determined on a DB-1701 (0.32 mm x 30 m, 0.25 microm) quartz capillary column. And the sample was extracted with ethanol by the ultrasonic assisted extraction. RESULT: The calibration curve of caryophyllene is liner over the range of 0.002-2.0 g x L (-1) (R2 = 1). The recovery was from 96.76% to 104.15%. CONCLUSION: The method is accurate, simple with a good reproducibility. It can be used to control the quality of E. fortune.