RESUMEN
The effect of bioflavonoids extracted from the bark of Pinus maritima, Pycnogenol (PYC), on gene expression of the proinflammatory cytokines interleukin-1beta (IL-1beta) and interleukin-2 (IL-2) were investigated in RAW 264.7 cells and Jurkat E6.1 cells, respectively. PYC exerted strong scavenging activities against reactive oxygen species (ROS) generated by H2O2 in RAW 264.7. In situ ELISA, immunoblot analysis, and competitive RT-PCR demonstrated that pretreatment of LPS-stimulated RAW 264.7 cells with PYC dose-dependently reduced both the production of IL-1beta and its mRNA levels. Furthermore, in the same cells, PYC blocked the activation of nuclear factor kappaB (NF-kappaB) and activator protein-1 (AP-1), two major transcription factors centrally involved in IL-1beta gene expression. Concordantly, pretreatment of the cells with PYC abolished the LPS-induced IkappaB degradation. We also investigated the effect of PYC on IL-2 gene expression in phorbol 12-myristate 13acetate plus ionomycin (PMA/Io)-stimulated human T-cell line Jurkat E6.1. PYC inhibited the PMA/Io-induced IL-2 mRNA expression. However, as demonstrated in a reporter gene assay system, the mechanism of IL-2 gene transcriptional regulation by PYC was different from the regulation of IL-1beta. PYC inhibited both NF-AT and AP-1 chloramphenicol acetyltransferase (CAT) activities in transiently transfected Jurkat E6.1, but not NF-kappaB CAT activity. We also found that PYC can destabilize PMA/Io-induced IL-2 mRNA by posttranscriptional regulation. All these results suggest that bioflavonids can be useful therapeutic agents in treating many inflammatory, autoimmune, and cardiovascular diseases based on its diverse action mechanisms.
Asunto(s)
Antioxidantes/farmacología , Flavonoides/farmacología , Depuradores de Radicales Libres/farmacología , Proteínas I-kappa B , Interleucina-1/biosíntesis , Interleucina-2/biosíntesis , Pinus/química , Corteza de la Planta/química , Animales , Antioxidantes/aislamiento & purificación , Cloranfenicol O-Acetiltransferasa/biosíntesis , Proteínas de Unión al ADN/metabolismo , Flavonoides/aislamiento & purificación , Depuradores de Radicales Libres/aislamiento & purificación , Regulación de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Genes Reporteros , Humanos , Interleucina-1/genética , Interleucina-2/genética , Ionomicina/farmacología , Células Jurkat/efectos de los fármacos , Células Jurkat/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Inhibidor NF-kappaB alfa , Proteínas de Neoplasias/biosíntesis , Proteínas de Neoplasias/genética , Extractos Vegetales , ARN Mensajero/biosíntesis , ARN Mensajero/genética , ARN Neoplásico/biosíntesis , ARN Neoplásico/genética , Especies Reactivas de Oxígeno/metabolismo , Proteínas Recombinantes de Fusión/biosíntesis , Acetato de Tetradecanoilforbol/farmacología , Factor de Transcripción AP-1/biosíntesis , Factor de Transcripción AP-1/genética , Factores de Transcripción/metabolismo , TransfecciónRESUMEN
Currently, bioflavonoids have been known to have strong antioxidant capacities, and a variety of efforts have been made to identify the utilities of bioflavonoids in treating various diseases based on their antioxidant capacities. The effects of bioflavonoids extracted from the bark of Pinus maritima Pycnogenol (PYC) on free radical formation, activation of redox sensitive transcription factors, as well as interleukin-1 beta (IL-1 beta) production were investigated in murine macrophage cell lines. PYC exerted strong scavenging activities against reactive oxygen species generated either by H(2)O(2) or PMA in RAW 264.7 and IC-21 cells, respectively. In situ ELISA, immunoblot analysis, and competitive RT-PCR demonstrated that PYC pretreatment of LPS-stimulated RAW 264.7 cells dose-dependently reduced both the production of IL-1 beta and its mRNA levels. Furthermore, in the same cells, PYC blocked the activation of nuclear factor kappa B (NF-kappa B) and activator protein-1 (AP-1), two major transcription factors centrally involved in IL-1 beta gene expression. When RAW 264.7 cells were stimulated with LPS, the inhibitor protein I kappa B largely disappeared from cytosolic fractions. However, pretreatment of the cells with PYC abolished the LPS-induced I kappa B degradation. These results suggest that PYC can inhibit the expression of the proinflammatory cytokine IL-1 by regulating redox-sensitive transcription factors. This study may support the possibility that bioflavonoids including PYC can be used as antiinflammatory and immunosuppressive drugs based on their radical scavenging activities.
Asunto(s)
Flavonoides/farmacología , Interleucina-1/biosíntesis , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Animales , Línea Celular , ADN/metabolismo , Depuradores de Radicales Libres/farmacología , Interleucina-1/farmacología , Mediciones Luminiscentes , Macrófagos/metabolismo , Ratones , FN-kappa B/metabolismo , Extractos Vegetales/farmacología , ARN Mensajero/análisis , Factor de Transcripción AP-1/metabolismoRESUMEN
The purpose of this study was to determine the factors which were responsible for delaying early diagnosis and optimal management of rheumatoid arthritis (RA) in Korea. We interviewed 109 outpatients diagnosed as RA being treated by rheumatologists, and we eventually analyzed 98 patients' data. The median length of time from symptom onset to the first visit to a medical doctor, to diagnosis, and visiting a rheumatologist were 8 weeks, 23 weeks, and 42 months respectively. The subspecialist with whom the patients consulted with for the longest time before visiting a rheumatologist were an orthopaedic surgeon for 51 patients, a Chinese herbal doctor for 19 patients, and a pharmacist for 16 patients. For early diagnosis and optimal management of RA in Korea, we believe that it is necessary to reduce the use of unconventional medical services such as Chinese herbal medicine and nonprescribed medication, and to emphasize rheumatologic and rehabilitative care in the early stage.