Graduate Institute of Immunology, China Medical University, 91 Hsueh-Shih Rd, Taichung 40402, Taiwan.
BMC Genomics. 2010 Sep 24;11:513. doi: 10.1186/1471-2164-11-513.
Functional comparative genomic analysis of the cellular immunological effects of different anti-inflammatory phytocompounds is considered as a helpful approach to distinguish the complex and specific bioactivities of candidate phytomedicines. Using LPS-stimulated THP-1 monocytes, we characterize here the immunomodulatory activities of three single phytocompounds (emodin, shikonin, and cytopiloyne) and a defined phytocompound mixture extracted from Echinacea plant (BF/S+L/Ep) by focused DNA microarray analysis of selected immune-related genes.
Shikonin and emodin significantly inhibited the early expression (within 0.5 h) of approximately 50 genes, notably cytokines TNF-α, IL-1β and IL-4, chemokines CCL4 and CCL8, and inflammatory modulators NFATC3 and PTGS2. In contrast, neither cytopiloyne nor BF/S+L/Ep inhibited the early expression of these 50 genes, but rather inhibited most late-stage expression (~12 h) of another immune gene subset. TRANSPATH database key node analysis identified the extracellular signal-regulated kinase (ERK) 1/2 activation pathway as the putative target of BF/S+L/Ep and cytopiloyne. Western blot confirmed that delayed inactivation of the ERK pathway was indeed demonstrable for these two preparations during the mid-stage (1 to 4 h) of LPS stimulation. We further identified ubiquitin pathway regulators, E6-AP and Rad23A, as possible key regulators for emodin and shikonin, respectively.
The current focused DNA microarray approach rapidly identified important subgenomic differences in the pattern of immune cell-related gene expression in response to specific anti-inflammatory phytocompounds. These molecular targets and deduced networks may be employed as a guide for classifying, monitoring and manipulating the molecular and immunological specificities of different anti-inflammatory phytocompounds in key immune cell systems and for potential pharmacological application.
对不同抗炎植物化合物的细胞免疫效应进行功能比较基因组分析，被认为是区分候选植物药复杂和特定生物活性的有益方法。在这里，我们使用 LPS 刺激的 THP-1 单核细胞，通过对选定的免疫相关基因进行聚焦 DNA 微阵列分析，来描述三种单一植物化合物（大黄素、紫草素和细胞色素）和一种从紫锥菊植物中提取的特定植物化合物混合物（BF/S+L/Ep）的免疫调节活性。
紫草素和大黄素显著抑制了大约 50 个基因的早期表达（在 0.5 小时内），特别是细胞因子 TNF-α、IL-1β 和 IL-4、趋化因子 CCL4 和 CCL8，以及炎症调节剂 NFATC3 和 PTGS2。相比之下，细胞色素和 BF/S+L/Ep 均未抑制这些 50 个基因的早期表达，而是抑制了另一组免疫基因的晚期表达（约 12 小时）。TRANSPATH 数据库关键节点分析确定细胞外信号调节激酶（ERK）1/2 激活途径是 BF/S+L/Ep 和细胞色素的潜在靶点。Western blot 证实，在 LPS 刺激的中期（1 至 4 小时），这两种制剂确实可以使 ERK 途径延迟失活。我们进一步确定了泛素途径调节剂 E6-AP 和 Rad23A，分别为大黄素和紫草素的可能关键调节剂。
目前的聚焦 DNA 微阵列方法快速确定了特定抗炎植物化合物对免疫细胞相关基因表达模式的重要亚基因组差异。这些分子靶点和推断的网络可用于指导不同抗炎植物化合物在关键免疫细胞系统中的分子和免疫学特异性的分类、监测和操纵，以及潜在的药理学应用。
