RESUMEN
BACKGROUND: In the past several decades, Polygonum viviparum L. (PV) was reported to have antibacterial, antiulcer, antioxidant, antitumor, anti-inflammatory, and antiarthritic properties. The anti-inflammatory pathway was recently elucidated through cytosolic nuclear factor E2-related factor 2 (Nrf2) activation and heme oxygenase (HO)-1 protein expression. PV is a perennial herb and widely distributed in high-elevation mountain regions, such as the Tibetan Plateau. In Tibetan traditional medicine, PV is usually used to boost the blood circulation to dissipate blood stasis. Therefore, this study focused on how PV improves the vascular circulation and acts on vascular tissues. METHODS: In this study, we isolated aortas from Sprague-Dawley rats (male, weight about 250~350 g), and detected the effects of PV on phenylephrine (PE)-induced contraction and cyclic guanosine 3',5'-monophosphate (cGMP) formation using aortic rings. In addition, human umbilical vein endothelial cells (HUVECs) were used to exam nitric oxygen (NO) synthase (NOS) activity by directly measuring NO production in the culture medium. Endothelial (e) NOS phosphorylation, and cytosolic Nrf2 and HO-1 expressions were measured using a Western blot analysis. RESULTS: PV dose-dependently relaxed PE-induced contractions in endothelial-intact but not -denuded aorta. The concentration to produce 50% relaxation was 22.04±1.77 µg/ml. PV-induced vasorelaxation was markedly blocked by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME), an NOS inhibitor, methylene blue (MB), a guanylyl cyclase inhibitor, and hemoglobin, an NO scavenger. PV increased cGMP formation; however, this effect was also suppressed by co-pretreatment with l-NAME, MB, hemoglobin, and Ca2+-free medium. In HUVECs, PV increased NO formation, which was greatly attenuated by NOS inhibitors (L-NAME and L-NMMA) and by removing extracellular Ca2+ and chelating intracellular Ca2+ with BAPTA-AM. In addition, PV promoted eNOS phosphorylation, Nrf2 degradation, and HO-1 protein expression according to a Western blot analysis. CONCLUSIONS: The results suggest that PV possesses vasorelaxing action in an endothelium-dependent manner and works through activating Ca2+/calmodulin- dependent NO synthesis; when NO is released and then transferred to smooth muscle cells, NO activates guanylyl cyclase and increases cGMP formation, ultimately resulting in vasorelaxation. Thus, PV can be considered for application as a potential therapeutic approach for vascular-associated disorders.
Asunto(s)
Aorta Torácica/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Óxido Nítrico Sintasa/metabolismo , Extractos Vegetales/farmacología , Polygonum/química , Vasodilatación/efectos de los fármacos , Animales , Aorta Torácica/citología , Aorta Torácica/enzimología , Aorta Torácica/fisiología , GMP Cíclico/metabolismo , Células Endoteliales/enzimología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/enzimología , Humanos , Técnicas In Vitro , Masculino , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Óxido Nítrico/metabolismo , Fenilefrina/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Polygonum viviparum L. (PV) is a member of the family Polygonaceae and is widely distributed in high-elevation areas. It is used as a folk remedy to treat inflammation-related diseases. This study was focused on the anti-inflammatory response of PV against lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages. RESULTS: Treatment with PV did not cause cytotoxicity at 0-50 µg mL(-1) in RAW264.7 macrophages, and the IC(50) value was 270 µg mL(-1). PV inhibited LPS-stimulated nitric oxide (NO), prostaglandin (PG)E(2) , interleukin (IL)-1ß and tumour necrosis factor (TNF)-α release and inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 protein expression. In addition, PV suppressed the LPS-induced p65 expression of nuclear factor (NF)-κB, which is associated with the inhibition of IκB-α degradation. These results suggest that, among mechanisms of the anti-inflammatory response, PV inhibits the production of NO and these cytokines by down-regulating iNOS and COX-2 gene expression. Furthermore, PV can induce haem oxygenase (HO)-1 protein expression through nuclear factor E2-related factor 2 (Nrf2) activation. A specific inhibitor of HO-1, zinc(II) protoporphyrin IX, inhibited the suppression of iNOS and COX-2 expression by PV. CONCLUSION: These results suggest that PV possesses anti-inflammatory actions in macrophages and works through a novel mechanism involving Nrf2 actions and HO-1. Thus PV could be considered for application as a potential therapeutic approach for inflammation-associated disorders.
Asunto(s)
Hemo-Oxigenasa 1/biosíntesis , Inflamación/prevención & control , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Factor 2 Relacionado con NF-E2/efectos de los fármacos , Polygonum/química , Animales , Antiinflamatorios , Línea Celular , Supervivencia Celular/efectos de los fármacos , Ciclooxigenasa 2/genética , Inhibidores de la Ciclooxigenasa/farmacología , Inducción Enzimática/efectos de los fármacos , Inflamación/inducido químicamente , Macrófagos/enzimología , Ratones , Factor 2 Relacionado con NF-E2/fisiología , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/genética , Extractos Vegetales/farmacologíaRESUMEN
BACKGROUND: Doxorubicin (DOX) is an effective antineoplastic drug; however, clinical use of DOX is limited by its dose-dependent cardiotoxicity. It is well known that reactive oxygen species (ROS) play a vital role in the pathological process of DOX-induced cardiotoxicity. For this study, we evaluated the protective effects of guggulsterone (GS), a steroid obtained from myrrh, to determine its preliminary mechanisms in defending against DOX-induced cytotoxicity in H9C2 cells. METHODS: In this study, we used a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release measurements, and Hoechst 33258 staining to evaluate the protective effect of GS against DOX-induced cytotoxicity in H9C2 cells. In addition, we observed the immunofluorescence of intracellular ROS and measured lipid peroxidation, caspase-3 activity, and apoptosis-related proteins by using Western blotting. RESULTS: The MTT assay and LDH release showed that treatment using GS (1-30 µM) did not cause cytotoxicity. Furthermore, GS inhibited DOX (1 µM)-induced cytotoxicity in a concentration-dependent manner. Hoechst 33258 staining showed that GS significantly reduced DOX-induced apoptosis and cell death. Using GS at a dose of 10-30 µM significantly reduced intracellular ROS and the formation of MDA in the supernatant of DOX-treated H9C2 cells and suppressed caspase-3 activity to reference levels. In immunoblot analysis, pretreatment using GS significantly reversed DOX-induced decrease of PARP, caspase-3 and bcl-2, and increase of bax, cytochrome C release, cleaved-PARP and cleaved-caspase-3. In addition, the properties of DOX-induced cancer cell (DLD-1 cells) death did not interfere when combined GS and DOX. CONCLUSION: These data provide considerable evidence that GS could serve as a novel cardioprotective agent against DOX-induced cardiotoxicity.
Asunto(s)
Antineoplásicos/toxicidad , Doxorrubicina/toxicidad , Miocitos Cardíacos/efectos de los fármacos , Pregnenodionas/farmacología , Sustancias Protectoras/farmacología , Terpenos/química , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Humanos , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The study was to examine the effects of Sanguis draconis ethanol extract (SDEE) on streptozotocin (STZ)- and cytokine-induced ß-cell damage. In vitro, SDEE did not cause cytotoxicity below 200 µg/ml, and can prevent STZ (5mM)-induced cell death and apoptosis below 100 µg/ml on RIN-m5F cells. SDEE inhibits IL-1ß/IFN-γ-stimulated NO, TNF-α release, and iNOS expression. Furthermore, SDEE suppressed the IL-1ß/IFN-γ- or STZ-induced p65 expression of NF-κB, which is associated with inhibition of IκB-α degradation. In vivo, treatment of ICR mice with STZ (100 mg/kg, i.p. single injection) resulted in hyperglycemia and hypoinsulinemia, which was further evidenced by blood glucose and plasma insulin. The diabetogenic effects of STZ were completely prevented when mice were orally administered with SDEE for 3 weeks, however, the blood glucose and plasma insulin showed no significant change after SDEE administration alone. In addition, SDEE also can inhibit STZ-induced iNOS protein expression, pancreatic injury and lipid peroxidation. In conclusions, the molecular mechanism by which SDEE inhibits iNOS gene expression appears to involve the inhibition of NF-κB activation. These results suggest the possible therapeutic value of S. draconis and could be potentially developed into a novel drug for preventing the progression of diabetes mellitus.
Asunto(s)
Células Secretoras de Insulina/efectos de los fármacos , Células Secretoras de Insulina/patología , Insulinoma/tratamiento farmacológico , Interferón gamma/toxicidad , Interleucina-1beta/toxicidad , Extractos Vegetales/farmacología , Estreptozocina/toxicidad , Animales , Antibióticos Antineoplásicos/toxicidad , Antivirales/toxicidad , Glucemia , Western Blotting , Supervivencia Celular/efectos de los fármacos , Citosol/metabolismo , Hiperglucemia/inducido químicamente , Hiperglucemia/tratamiento farmacológico , Técnicas In Vitro , Células Secretoras de Insulina/metabolismo , Islotes Pancreáticos/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos ICR , Óxido Nítrico/metabolismo , Ratas , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVES: To elucidate a novel anti-inflammatory mechanism of myrrh against lipopolysaccharide (LPS)-induced inflammation. METHODS: RAW264.7 macrophages were cultured in DMEM and then cells were treated with LPS or LPS plus a myrrh methanol extract (MME) for 24h. The culture medium was collected for determination of nitric oxide (NO), prostaglandin (PG)E(2) , interleukin (IL)-1ß, and tumour necrosis factor (TNF)-α, and cells were harvested by lysis buffer for Western blot analysis. KEY FINDINGS: Our data showed that treatment with the MME (1â¼100µg/ml) did not cause cytotoxicity or activate haem oxygenase-1 (HO-1) protein synthesis in RAW264.7 macrophages. Furthermore, the MME inhibited LPS-stimulated NO, PGE(2) , IL-1ß and TNF-α release and inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein expression. Zn(II) protoporphyrin IX, a specific inhibitor of HO-1, blocked the inhibition of iNOS and COX-2 expression by the MME. CONCLUSIONS: These results suggest that among mechanisms of the anti-inflammatory response, the MME inhibited the production of NO, PGE(2) , IL-1ß and TNF-α by downregulating iNOS and COX-2 gene expression in macrophages and worked through the action of HO-1.
Asunto(s)
Antiinflamatorios/farmacología , Commiphora , Hemo-Oxigenasa 1/biosíntesis , Mediadores de Inflamación/metabolismo , Inflamación/metabolismo , Fitoterapia , Extractos Vegetales/farmacología , Animales , Antiinflamatorios/uso terapéutico , Inflamación/tratamiento farmacológico , Lipopolisacáridos , Macrófagos/efectos de los fármacos , Ratones , Extractos Vegetales/uso terapéuticoRESUMEN
BACKGROUND: Safflower, whose botanic name is Carthamus tinctorius L., is a member of the family Compositae or Asteraceae. Carthamus yellow (CY) is the main constituent of safflower and is composed of safflomin A and safflomin B. Dried safflower petals are used in folk medicine and have been shown to invigorate blood circulation, break up blood stasis, and promote menstruation. In addition, dried safflower petals contain yellow dyes that are used to color food and cosmetics. In this study, we investigated the effects of dried safflower petals aqueous extracts (SFA) and CY on lipopolysaccharide (LPS)-induced inflammation using RAW264.7 macrophages. RESULTS: Our data showed that treatment with SFA (1-1000 microg mL(-1)) and CY (1-2000 microg mL(-1)) does not cause cytotoxicity in cells. SFA and CY inhibited LPS-stimulated nitric oxide (NO), prostaglandin E(2) (PGE(2)), and interleukin 1ß (IL-1ß) release, through attenuation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression. Further, SFA and CY suppressed the LPS-induced phosphorylation of nuclear factor-κB, which was associated with the inhibition of IκB-α degradation. CONCLUSION: These results suggest that SFA and CY provide an anti-inflammatory response through inhibiting the production of NO and PGE(2) by the downregulation of iNOS and COX-2 gene expression. Thus safflower petals have the potential to provide a therapeutic approach to inflammation-associated disorders.
Asunto(s)
Carthamus tinctorius/química , Mediadores de Inflamación/metabolismo , Inflamación/tratamiento farmacológico , Macrófagos/efectos de los fármacos , Fenoles/uso terapéutico , Fitoterapia , Extractos Vegetales/uso terapéutico , Animales , Ciclooxigenasa 2/metabolismo , Flores/química , Proteínas I-kappa B/metabolismo , Inflamación/metabolismo , Lipopolisacáridos , Ratones , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenoles/aislamiento & purificación , Fenoles/farmacología , Fosforilación , Extractos Vegetales/farmacologíaRESUMEN
Brazilin, the main constituent of Caesalpinia sappan L., is a natural red pigment that has been reported to possess anti-inflammatory properties. This study aimed to identify a novel anti-inflammatory mechanism of brazilin. We found that brazilin did not cause cytotoxicity below 300 microM, and activated heme oxygenase-1 (HO-1) protein synthesis in a concentration-dependent manner at 10-300 microM in RAW264.7 macrophages without affecting mRNA transcription of HO-1. Additionally, brazilin increased bilirubin production and HO-1 activity in RAW264.7 macrophages. In lipopolysaccharide (LPS)-stimulated macrophages, brazilin suppressed the release of nitric oxide (NO), prostaglandin E(2) (PGE(2)), interleukin (IL)-1beta and tumor necrosis factor-alpha (TNF-alpha), and reduced the expression of inducible nitric oxide synthase (iNOS). A specific inhibitor of HO-1, Zn(II) protoporphyrin IX, blocked the suppression of NO production, cytokines release and iNOS expression by brazilin. These results suggest that brazilin possesses anti-inflammatory actions in macrophages and works through a novel mechanism involving the action of HO-1.