Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8572, Japan.
Cytotechnology. 2011 Mar;63(2):181-90. doi: 10.1007/s10616-010-9333-6. Epub 2011 Feb 18.
Fulvic acid (FA) is class of compounds of humic substances formed through the degradation of organic substances by chemical and biological processes. FA has been utilized in traditional Chinese medicine and possesses various pharmacological properties. Previously, we reported that FA extracted from solubilized excess sludge (SS-FA) had an inhibitory effect on β-hexosaminidase release in human leukemia basophilic (KU812) cells. In this study, we investigated the effects of SS-FA on the immediate-type allergic reaction and studied its possible mechanisms of action in KU812 cells following activation with phorbol myristate acetate (20 nmol L(-1)) plus calcium ionophore A23187 (1 μmol L(-1)) (PMACI). The inhibitory effect of SS-FA on degranulation in PMACI-stimulated KU812 cells was examined using histamine release assay. SS-FA significantly decreased the histamine release in KU812 cells at concentrations of 0.1-10.0 μg mL(-1). To gain more information regarding the mechanism of the suppression of degranulation following SS-FA treatment, microarray was conducted to determine which genes were differentially expressed in response to SS-FA in PMACI-activated KU812 cells. From a total of 201 genes in the DNA chip, 28 genes were up-regulated and 173 genes were down-regulated in cells pretreated with SS-FA for 15 min and stimulated with PMACI. From the 71 genes that showed more than two fold change in expression, 16 genes were significantly down-regulated that were subjected to hierarchical clustering. SS-FA affected the expression of genes that were involved in the following pathways: signal transduction, cytokine-cytokine receptor interaction, immune response, cell adhesion molecules and IgE receptor β subunit response.
富里酸(FA)是腐殖质化合物的一类,通过化学和生物过程降解有机物质形成。FA 已被用于中药,并具有多种药理特性。此前,我们报道了从溶解过剩污泥中提取的 FA(SS-FA)对人白血病嗜碱性细胞(KU812)β-己糖胺酶释放具有抑制作用。在这项研究中,我们研究了 SS-FA 对即刻型过敏反应的影响,并研究了其在 KU812 细胞经佛波醇肉豆蔻酸酯(20 nmol L(-1))加钙离子载体 A23187(1 μmol L(-1))(PMACI)激活后可能的作用机制。使用组氨酸释放测定法检测 SS-FA 对 PMACI 刺激的 KU812 细胞脱粒的抑制作用。SS-FA 在 0.1-10.0 μg mL(-1)浓度下显著降低 KU812 细胞中的组氨酸释放。为了获得更多关于 SS-FA 处理后脱粒抑制机制的信息,进行了微阵列分析以确定在 PMACI 激活的 KU812 细胞中 SS-FA 处理后差异表达的基因。在 DNA 芯片中的 201 个基因中,在 SS-FA 预处理 15 分钟并用 PMACI 刺激的细胞中,有 28 个基因上调,173 个基因下调。在表达变化超过两倍的 71 个基因中,有 16 个基因明显下调,这些基因进行了层次聚类。SS-FA 影响参与以下途径的基因表达:信号转导、细胞因子-细胞因子受体相互作用、免疫反应、细胞粘附分子和 IgE 受体β亚基反应。