Ethanolic extract and water-soluble polysaccharide from Chaenomeles speciosa fruit modulate lipopolysaccharide-induced nitric oxide production in RAW264.7 macrophage cells
Ethanolic extract and water-soluble polysaccharide from Chaenomeles speciosa fruit modulate lipopolysaccharide-induced nitric oxide production in RAW264.7 macrophage cells
复制标题
木瓜果实的乙醇提取物和水溶性多糖调节脂多糖诱导的 RAW264.7 巨噬细胞中一氧化氮的产生
DOI：
10.1016/j.jep.2012.09.042
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发表时间：
2012-11-21
影响因子：
5.4
通讯作者：
Huang, Zebo
中科院分区：
文献类型：
--
作者：
Zhu, Qing;Liao, Chaolin;Huang, Zebo
民族药理学相关性：木瓜果实在中医药中广泛用于治疗与炎症反应有关的疾病。本研究旨在鉴定木瓜果实的抗炎和免疫调节活性成分，并探讨其可能的作用机制。材料与方法：从木瓜干果中提取乙醇提取物及其正己烷、氯仿、乙酸乙酯和正丁醇部位，以及水溶性多糖。用脂多糖（LPS）诱导小鼠巨噬细胞样RAW264.7细胞作为炎症细胞模型。Griess法测定细胞中一氧化氮的产生，MTT法测定细胞活力。通过荧光激活细胞分选术评价细胞凋亡。炎症相关基因的相对定量分析通过real-timePCR.Results：LPS诱导的RAW264.7细胞中一氧化氮的产生被显着抑制乙酸乙酯馏分（EAF）在200-800 μ g/ml，而木瓜多糖（CPS）促进一氧化氮的产生在250-750 μ g/ml，无论是单独或在添加剂的方式与LPS。在上述有效浓度下，EAF和CPS均未引起明显的细胞毒性和细胞凋亡。EAF能显著降低LPS诱导的iNOS mRNA水平的上调，但对IFN-γ和G-CSF的诱导无显著影响; CPS能显著降低LPS诱导的TNF-α、IFN-γ和G-CSF的基因诱导。CPS是通过细胞因子如TNF-α、IFN-γ和G-CSF产生一氧化氮的激活剂。这些结果证明了木瓜果实的乙醇和水提取物的治疗效果，木瓜果实是一种用于健康维护和疾病治疗的传统食用药物。(C)2012爱思唯尔爱尔兰有限公司保留所有权利。
Ethnopharmacological relevance: Chaenomeles speciosa fruits have been widely used in traditional Chinese medicine for treatment of diseases related to inflammatory reaction. This study aims to identify anti-inflammatory and immunomodulatory components of Chaenomeles speciosa fruit and unravel their potential mechanisms.Materials and methods: Ethanolic extract and its n-hexane, chloroform, ethyl acetate and n-butanol fractions, as well as water-soluble polysaccharide, were prepared from dry fruits of Chaenomeles speciosa. The mouse macrophage-like RAW264.7 cells were induced by lipopolysaccharide (LPS) and used as an inflammatory cell model. Production of nitric oxide in the cells was determined by the Griess assay, and cell viability was tested by the MTT method. Cellular apoptosis was evaluated by fluorescence-activated cell sorting. Relative quantification of inflammation-related genes was analyzed by real-time PCR.Results: LPS-induced production of nitric oxide in RAW264.7 cells was significantly inhibited by the ethyl acetate fraction (EAF) at 200-800 mu g/ml, while Chaenomeles speciosa polysaccharide (CPS) promoted nitric oxide production at 250-750 mu g/ml either alone or in an additive fashion with LPS. Both EAF and CPS did not provoke noticeable cytotoxicity and apoptosis at the above effective concentrations. EAF significantly reduced LPS-induced upshift of iNOS mRNA level but showed no significant effect on the induction of IFN-gamma and G-CSF, while CPS reduced the gene induction of TNF-alpha, IFN-gamma and G-CSF by LPS.Conclusions: EAF was able to inhibit nitric oxide production by reducing LPS-induced upshift of iNOS mRNA level. CPS was an activator of nitric oxide production through cytokines such as TNF-alpha, IFN-gamma and G-CSF. These results demonstrate the therapeutic effects of both ethanolic and aqueous extracts of Chaenomeles speciosa fruit, a traditional edible medicine used in health maintenance and disease treatment. (C) 2012 Elsevier Ireland Ltd. All rights reserved.