ABSTRACT
BACKGROUND: Psoriasis is a common chronic inflammatory skin disease. Keratinocytes hyperproliferation and excessive inflammatory response contribute to psoriasis pathogenesis. The agents able to attenuate keratinocytes hyper-proliferation and excessive inflammatory response are considered to be potentially useful for psoriasis treatment. Daphnetin exhibits broad bioactivities including anti-proliferation and anti-inflammatory. This study aims to evaluate the anti-psoriatic potential of daphnetin in vitro and in vivo, and explore underlying mechanisms. METHODS: HaCaT keratinocytes was stimulated with the mixture of IL-17A, IL-22, oncostatin M, IL-1α, and TNF-α (M5) to establish psoriatic keratinocyte model in vitro. Cell viability was measured using Cell Counting Kit-8 (CCK-8). Quantitative Real-Time PCR (qRT-PCR) was performed to measure the mRNA levels of hyperproliferative marker gene keratin 6 (KRT6), differentiation marker gene keratin 1 (KRT1) and inflammatory factors IL-1ß, IL-6, IL-8, TNF-α, IL-23A and MCP-1. Western blotting was used to detect the protein levels of p65 and p-p65. Indirect immunofluorescence assay (IFA) was carried out to detect p65 nuclear translocation. Imiquimod (IMQ) was used to construct psoriasis-like mouse model. Psoriasis severity (erythema, scaling) was scored based on Psoriasis Area Severity Index (PASI). Hematoxylin and eosin (H&E) staining was performed to examine histological change in skin lesion. The expression of inflammatory factors including IL-6, TNF-α, IL-23A and IL-17A in skin lesion was measured by qRT-PCR. RESULTS: Daphnetin attenuated M5-induced hyperproliferation in HaCaT keratinocytes. M5 stimulation significantly upregulated mRNA levels of IL-1ß, IL-6, IL-8, TNF-α, IL-23A and MCP-1. However, daphnetin treatment partially attenuated the upregulation of those inflammatory cytokines. Daphnetin was found to be able to inhibit p65 phosphorylation and nuclear translocation in HaCaT keratinocytes. In addition, daphnetin significantly ameliorate the severity of skin lesion (erythema, scaling and epidermal thickness, inflammatory cell infiltration) in IMQ-induced psoriasis-like mouse model. Daphnetin treatment attenuated IMQ-induced upregulation of inflammatory cytokines including IL-6, IL-23A and IL-17A in skin lesion of mice. CONCLUSIONS: Daphnetin was able to attenuate proliferation and inflammatory response induced by M5 in HaCaT keratinocytes through suppression of NF-κB signaling pathway. Daphnetin could ameliorate the severity of skin lesion and improve inflammation status in IMQ-induced psoriasis-like mouse model. Daphnetin could be an attractive candidate for future development as an anti-psoriatic agent.
Subject(s)
Humans , Animals , Mice , Rabbits , Psoriasis/chemically induced , Psoriasis/drug therapy , Umbelliferones/pharmacology , Adjuvants, Immunologic/adverse effects , Imiquimod/adverse effects , Inflammation/drug therapy , Anti-Inflammatory Agents/pharmacology , Keratinocytes , Cell Proliferation , Mice, Inbred BALB CABSTRACT
INTRODUCTION: Justicia pectoralis Jacq. (Acanthaceae) is medicinal plant species commonly used in Cuba for the treatment of nervous disorders because of its sedative effect. Coumarin is one of its main active phytochemicals present in the extracts obtained from this plant and used as analytic marker in quality control. On the other hand, this compound contributes to the sedative effect attributed to this plant. OBJECTIVE: to evaluate the influence of harvest time on the coumarin and umbelliferone (7-hydroxycoumarin) in Justicia pectoralis extracts. METHODS: the experiment lasted two years. The harvest was performed at 4, 6 and 8 months after planting. Aqueous and hydroalcohol extracts were produced and the coumarin and umbelliferone contents were determined by high resolution liquid chromatography. RESULTS: the achieved results showed the presence of coumarin and umbelliferone in both extracts. Both methods can be used for the extraction of these components from the plant, although in the case of umbelliferone, the best extraction results were achieved by using aqueous extract. In both cases, the recovery percentages were more than 98 percent. This study confirmed that the harvest time significantly influences on the coumarin and umbelliferone contents. CONCLUSIONS: the best results are observed in the first two harvests (4 and 6 months at summer time), which indicates that the industry should process the vegetal material in these two periods of the year(AU)
INTRODUCCIÓN: Justicia pectoralis Jacq. (Acanthaceae) es una planta medicinal comúnmente usada en Cuba para el tratamiento de enfermedades nerviosas por su efecto sedante. La cumarina es uno de los fitocomponentes mayoritarios en los extractos obtenidos con esta planta y empleado como marcador analítico en los controles de calidad. Por otro lado, este componente contribuye con el efecto sedante atribuido a esta planta. OBJETIVO: evaluar la influencia del tiempo de cosecha sobre el contenido de cumarina y umbelliferona (7 hidroxicumarina) en extractos de Justicia pectoralis. MÉTODOS: se desarrolló el experimento durante 2 años. Se realizaron las cosechas a los 4, 6 y 8 meses de plantada. Se elaboraron extractos acuosos e hidroalcohólicos y se determinó el contenido de cumarina y umbelliferone por cromatografía líquida de alta resolución. RESULTADOS: se mostró la presencia de cumarina y umbelliferona en ambos extractos. Además, en el caso de la umbelliferona, los mejores resultados se alcanzaron al aplicar extracción acuosa. En ambos casos, los por cientos de recobrados fueron superiores al 98 por ciento. Se confirmó que el tiempo de cosecha influyó significativamente sobre el contenido de cumarina y umbelliferona. CONCLUSIONES: los mejores resultados se obtienen en la primeras dos cosechas (4 y 6 meses que coincide con el verano), lo que sugiere que el material de la planta debe procesarse por la industria en esos periodos del año(AU)
Subject(s)
Umbelliferones/pharmacology , Coumarins/pharmacology , Cuba , TiliaABSTRACT
The antiproliferative and cytotoxic effects of closely related coumarins, including esculetin, scopoletin and umbelliferone against human T, B lymphoid, myeloid, erythroid and mastocytoma cell lines, were studied in vitro. Esculetin was found to inhibit, dose- dependently, the proliferation of most cell lines. Myeloblastic leukemia, KG-1, was the most sensitive to the effect of esculetin. The Lc 50 was 24.7 micro M. Other cell lines were also susceptible to the effect of esculetin. CEM demonstrated an Lc 50 of 35.2 micro M, and EBV-transformed cell lines demonstrated an Lc 50 of 41.7 and 46.9 micro M. Some cell lines were quite resistant and include T-cell line Ke 37 and the erythroblastic leukemia K562. Interestingly, CTLL16, a crytotoxic normal T-cell line which is dependent on its growth on IL-2, did not show any appreciable cytotoxic effect to esculetin even when the cell had undergone 2 cycles of cell division. Mastocytoma showed in vitro stimulation by esculetin at low concentrations and by scopletin and umbelliferone at high concentrations, in vivo studies indicated that esculetin, scopoletin and umbelliferone were potent inhibitors of bone marrow mitosis. The results were discussed based on available data relating the effects of esculetin on leukotrienes and leukotrienes role in cell growth