Department of Oral Microbiology and Immunology, Dental Research Institute, and BK21 Program, School of Dentistry, Seoul National University, Seoul 110-749, Republic of Korea.
J Ethnopharmacol. 2010 Mar 2;128(1):198-205. doi: 10.1016/j.jep.2010.01.011. Epub 2010 Jan 14.
Armillariella mellea is an edible mushroom that has been traditionally used as an alternative medicine in many countries because of its anti-microbial and anti-cancer effects.
In this study, we examined the ability of Armillariella mellea to induce the expression of intercellular adhesion molecule (ICAM)-1, an important cellular adhesion molecule for the recruitment of immune cells to regional inflammatory sites.
A human monocytic cell line, THP-1 or human peripheral blood mononuclear cells (PBMC) were stimulated with Armillariella mellea extract (AME) and subjected to flow cytometry to examine the expression of ICAM-1 protein on the cell surface. Steady-state mRNA level of ICAM-1 was determined by real-time reverse transcription-polymerase chain reaction. The phosphorylation of JNK protein was examined by Western blot analysis using antibodies specific for non-phosphorylated and phosphorylated forms of JNK. For the analysis of transcription factors regulating ICAM-1 transcription, the nuclear fraction was extracted from AME-treated THP-1 cells and subjected to electrophoretic mobility shift assay.
AME induced expression of ICAM-1 and its mRNA in THP-1 cells in dose- and time-dependent manners. AME-induced ICAM-1 expression was also observed on CD14-positive monocytes in human PBMC. Interestingly, AME-induced ICAM-1 production was inhibited by the specific inhibitors of reactive oxygen species (ROS) and JNK, whereas no inhibitory effect was observed when inhibitors of ERK, p38 kinase, phosphatidylinositol 3-kinase, or protein kinase C were used. Concomitantly, AME increased phosphorylation of JNK in a time-dependent fashion. DNA binding activities of NF-kappaB, AP-1, SP-1, and STAT-1 were increased by AME treatment.
These results suggest that AME induces ICAM-1 expression in human monocytic cells through ROS/JNK-dependent signaling pathways leading to the activation of NF-kappaB, AP-1, SP-1, and STAT-1 transcription factors.
药用真菌蜜环菌能诱导细胞间黏附分子-1 的表达
药用真菌蜜环菌是一种可食用的蘑菇，由于其具有抗微生物和抗癌作用，在许多国家已被传统地用作替代药物。
在这项研究中，我们研究了蜜环菌提取物（AME）诱导细胞间黏附分子-1（ICAM-1）表达的能力，ICAM-1 是一种重要的细胞黏附分子，可将免疫细胞募集到局部炎症部位。
用 AME 刺激人单核细胞系 THP-1 或人外周血单核细胞（PBMC），并用流式细胞术检测细胞表面 ICAM-1 蛋白的表达。通过实时逆转录-聚合酶链反应测定 ICAM-1 的稳定 mRNA 水平。用特异性针对 JNK 非磷酸化和磷酸化形式的抗体通过 Western blot 分析检测 JNK 蛋白的磷酸化。为了分析调节 ICAM-1 转录的转录因子，从 AME 处理的 THP-1 细胞中提取核部分，并进行电泳迁移率变动分析。
AME 以剂量和时间依赖的方式诱导 THP-1 细胞中 ICAM-1 的表达和其 mRNA。AME 还可诱导人 PBMC 中 CD14 阳性单核细胞中 ICAM-1 的表达。有趣的是，AME 诱导的 ICAM-1 产生可被 ROS 和 JNK 的特异性抑制剂抑制，而当使用 ERK、p38 激酶、磷脂酰肌醇 3-激酶或蛋白激酶 C 的抑制剂时则没有抑制作用。同时，AME 以时间依赖的方式增加 JNK 的磷酸化。AME 处理后 NF-κB、AP-1、SP-1 和 STAT-1 的 DNA 结合活性增加。
这些结果表明，AME 通过 ROS/JNK 依赖性信号通路诱导人单核细胞中 ICAM-1 的表达，从而激活 NF-κB、AP-1、SP-1 和 STAT-1 转录因子。
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