RESUMEN
Chronic inflammation is a fundamental symptom of many diseases. Catechin possesses anti-oxidant and anti-inflammatory properties. However, the mechanism of catechin to prevent inflammation in 3T3-L1 adipocytes caused by TNF-α remains unknown. Therefore, the effects of catechin on the gene expression of cytokines and the activation of cell signals in TNF-α induced 3T3-L1 adipocytes were investigated. The effects of catechin on adipogenesis and cell viability were detected by Oil Red O staining and CCK-8 assay, respectively. The genes expression of cytokines was determined by real-time RT-PCR. The expression of NF-κB, AMPK, FOXO3a and SIRT1 on translation level was determined by western blotting analysis. The results demonstrated that catechin significantly enhanced adipogenesis and cell viability. catechin inhibited the gene expression of pro-inflammatory cytokines including IL-1α, IL-1ß, IL-6, IL-12p35, and inflammatory enzymes including iNOS and COX-2, but enhanced the gene expression of anti-inflammatory cytokines including IL-4 and IL-10. Catechin also inhibited the activation of NF-κB, AMPK, FOXO3a and SIRT1, but increased the phosphorylation level of the above factors. All these results indicated that as a potential therapeutic strategy catechin has the ability of attenuating inflammatory response triggered by TNF-α through signaling cascades involved in inflammation and cytokines.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Adipocitos/efectos de los fármacos , Catequina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Inflamación/prevención & control , Sirtuina 1/metabolismo , Factor de Necrosis Tumoral alfa/toxicidad , Células 3T3-L1 , Proteínas Quinasas Activadas por AMP/genética , Adipocitos/inmunología , Adipocitos/metabolismo , Adipogénesis , Animales , Citocinas/metabolismo , Inflamación/inducido químicamente , Inflamación/metabolismo , Inflamación/patología , Ratones , FN-kappa B/genética , FN-kappa B/metabolismo , Transducción de Señal , Sirtuina 1/genéticaRESUMEN
BACKGROUND: The use of plant essential oils as pharmaceuticals is a fast-growing market especially in China. Throughout the 20th century, a rapid increase took place in the use of many essential oil-derived products in the medicinal industry as nutraceuticals, medicinal supplements, and pharmaceuticals. PURPOSE: The objective of this study was to explore the chemical composition of Croton crassifolius essential oil as well as its potential anti-tumour properties and related anti-proliferative, autophagic, and apoptosis-inducing effects. METHODS: Supercritical CO2 fluid extraction technology was used to extract CCEO and the chemical constituents of the essential oil were identified by comparing the retention indices and mass spectra data taken from the NIST library with those calculated based on the C7-C40 n-alkanes standard. The cytotoxic activity and anti-proliferative effects of CCEO were evaluated against five cancer cell lines and one normal human cell line via CCK-8 assays. In addition, flow cytometry was used to detect cell cycle arrest. The efficacy of CCEO treatments in controlling cancer cell proliferation was assessed by cell cycle analysis, clonal formation assays, RT-qPCR, and western blot analysis. Autophagic and apoptosis-inducing effects of oils and the associated molecular mechanisms were assessed by flow cytometry, cell staining, reactive oxygen species assays, RT-qPCR, and western blot analysis. CONCLUSION: Forty compounds representing 92.90% of the total oil were identified in CCEO. The results showed that CCEO exerted a measurable selectivity for cancer cell lines, especially for A549 with the lowest IC50 value of 25.00 ± 1.62 µg/mL. Assessment of the anti-proliferative effects of CCEO on A549 cells showed that the oil inhibited cell proliferation and colony formation in a dose- and time-dependent manner. Investigation of the molecular mechanisms of cell cycle regulation confirmed that the oil arrested A549 cells in G2/M phase by decreasing the expression of cyclin B1-CDK1 and cyclin A-CDK1 and increasing the expression of cyclin-dependent kinase inhibitor (CKI) P21 at both the transcriptional and translational levels. Autophagy staining assays and western blot analysis revealed that CCEO promoted the formation of autophagic vacuoles in A549 cells and increased the expression of autophagy-related proteins beclin-1 and LC3-II in a dose-dependent manner. A series of apoptosis analyses indicated that CCEO induces apoptosis through a mitochondria-mediated intrinsic pathway. This study revealed that CCEO is a promising candidate for development into an anti-tumour drug of the future.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Cromatografía con Fluido Supercrítico/métodos , Croton/química , Aceites Volátiles/química , Células A549 , Antineoplásicos Fitogénicos/química , Autofagia/efectos de los fármacos , Beclina-1/metabolismo , Proteína Quinasa CDC2/metabolismo , Dióxido de Carbono/química , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Aceites Volátiles/análisis , Raíces de Plantas/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Chemical investigation of the anomalous fruits of Gleditsia sinensis led to the isolation and identification of a new triterpenoid saponin, 3-O-ß-D-xylopyranosyl-(1 â 2)-α-L-arabinopyranosyl-(1 â 6)-ß-D-glucopyranosyl oleanolic acid 28-O-ß-D-xylopyranosyl-(1 â 4)-α-L-rhamnopyrano--syl-(1 â 4)-ß-D-xylopyranosyl-(1 â 4)-α-L-rhamnopyranosyl-(1 â 3)-ß-D-glucopyranosyl ester (1), along with other nine known compounds (2-10). All the isolates from this species were reported for the first time. The structure of Compound 1 was determined by a detailed analysis using various analytical techniques, including 1D and 2D NMR. In vitro antiproliferative activities of Compound 1 on MCF-7 and Hep-G2 tumor cell lines were evaluated. IC50 values against the two cell lines were 9.5 and 11.6 µM, respectively.