Effects of blue honeysuckle (Lonicera caerulea L.) extract on lipopolysaccharide-induced inflammation in vitro and in vivo
Effects of blue honeysuckle (Lonicera caerulea L.) extract on lipopolysaccharide-induced inflammation in vitro and in vivo
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DOI：
10.1016/j.exer.2005.10.024
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发表时间：
2006-05-01
影响因子：
3.4
通讯作者：
Ohno, S
中科院分区：
文献类型：
--
作者：
Jin, XH;Ohgami, K;Ohno, S
本研究的目的是研究含有高含量酚类化合物的蓝金银花提取物（BHE）对内毒素诱导的葡萄膜炎（EIU）的影响。将雄性刘易斯大鼠随机分为5组，每组14只（8只收集房水，6只进行组织学检查）。通过足垫注射脂多糖（LPS）诱导ElU。注射LPS后立即静脉注射1、10或100 mg BHE。于注射LPS后24 h收集眼房水，测定房水中浸润细胞数、蛋白质浓度、一氧化氮（NO）、肿瘤坏死因子（TNF）-α和前列腺素（PG）-E2水平。部分眼球摘出进行组织学检查和免疫组化分析。用抗活化核因子（NF）-κ B的单克隆抗体进行免疫组织化学染色，以评价BHE对NF-κ B活化的影响。为了进一步阐明抗炎作用，在存在或不存在BHE及其主要酚类物质、矢车菊素3-葡糖苷（M）、矢车菊素3-芸香糖苷（C3 R）、绿原酸（CA）的情况下，用LPS刺激RAW 264.7细胞（小鼠巨噬细胞系）。Western blot检测诱导型一氧化氮合酶（iNOS）和环氧合酶2（考克斯2）的表达。BHE治疗显著减少炎性细胞浸润，降低房水中蛋白浓度，降低NO、TNF-α和PGE 2水平，并改善眼组织的组织学状态。在LPS注射后3 h，经BRE处理的虹膜睫状体中活化的NF-κ B阳性细胞的数量较低。BHE能显著抑制LPS刺激的RAW264.7细胞培养液中NO、PGE 2和TNF-α的产生，并呈剂量依赖性抑制iNOS和考克斯-2的表达。C3 G、C3 R和CA对炎症介质水平和iNOS、考克斯-2表达无抑制作用或抑制作用较弱。这些结果表明，BHE通过抑制NF-κ B依赖性信号通路和随后的促炎介质的产生来减轻EIU眼的炎症程度。(c)2005爱思唯尔有限公司保留所有权利。
The aim of the present study was to investigate the effects of blue honeysuckle extract (BHE), which contains high level of phenolic compounds, on endotoxin-induced uveitis (EIU). Male Lewis rats were randomly divided into 5 groups with 14 rats in each (eight rats for collection of aqueous humor, six rats for histologic examination). ElU was induced by a footpad injection of lipopolysaccharide (LPS). 1, 10, or 100 mg of BHE was injected intravenously immediately after LPS injection. The aqueous humor was collected at 24 h after LPS injection, the number of infiltrating cells, protein concentration, nitric oxide (NO), tumor necrosis factor (TNF)-alpha, and prostaglandin (PG)-E2 levels in the aqueous humor were determined. Some eyes were enucleated for histologic examination and immunohistochemical analysis. Immunohistochemical staining with a monoclonal antibody against activated nuclear factor (NF)-kappa B was performed to evaluate the effect of BHE on NF-kappa B activation. To further clarify the anti-inflammatory effect, RAW264.7 cells (a mouse macrophage cell line) were stimulated with LPS in the presence or absence of BHE and its major phenolics, cyanidin 3-glucoside (M), cyanidin 3-rutinoside (C3R), chlorogenic acid (CA). Expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by Western blot method. BHE treatment significantly reduced the inflammatory cell infiltration, the protein concentration, the levels of NO, TNF-alpha and PGE2 in the aqueous humor and improved histologic status of the ocular tissue. The number of activated NF-kappa B-positive cells was lower in the iris-ciliary body treated with BRE at 3 h after LPS injection. BHE significantly suppressed the production of NO, PGE2 and TNF-alpha in the culture medium as well as the expression of iNOS and COX-2 by LPS-stimulated RAW264.7 cells in a dose-dependent fashion. C3G, C3R and CA showed no or weak inhibitory effects on the level of inflammatory mediators and the expression of iNOS and COX-2. These results suggest that BHE attenuates the degree of inflammation in eyes with EIU by inhibiting the NF-kappa B dependent signaling pathway and the subsequent production of proinflammatory mediators. (c) 2005 Elsevier Ltd. All rights reserved.