Microarray analysis of gene expression profile by treatment of Cinnamomi Ramulus in lipopolysaccharide-stimulated BV-2 cells
Microarray analysis of gene expression profile by treatment of Cinnamomi Ramulus in lipopolysaccharide-stimulated BV-2 cells
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DOI：
10.1016/j.gene.2009.04.024
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发表时间：
2009-08-15
期刊：
影响因子：
3.5
通讯作者：
Lim, Sabina
中科院分区：
文献类型：
--
作者：
Hwang, Se-Hee;Choi, Yeong Gon;Lim, Sabina
据报道，肉桂Ramulus （Cinnamomi Ramulus， CR）的成分通过抑制诱导型一氧化氮合成（iNOS）和环氧合酶-2 （COX-2）的表达以及抑制中枢神经系统（CNS）和外周细胞中一氧化氮（NO）的产生而具有抗炎作用。本研究通过微阵列分析研究了CR对脂多糖（LPS）刺激的BV-2小胶质细胞基因表达和相关通路的影响。小胶质细胞在几种炎症介导的神经退行性疾病的过程中起重要作用。激活的小胶质细胞可以产生各种促炎细胞因子、趋化因子和毒性介质，这些物质可以启动或放大中枢神经系统的炎症反应。在本研究中，阴性对照组在正常培养基中培养，阳性对照组用1 μ g/ml LPS激活，CR组在LPS刺激前先用10 μ g/ml CR处理。截止值为1.5倍的表达变化，在lps刺激的BV-2细胞中发现341个基因表达上调，包括促炎细胞因子、趋化因子和转录因子（补充表2）。CR降低了lps诱导的炎症基因Ccl5、Cd80、Cxcl10、Grin2a、Ifi203、Ifit1、Il1 α、116、Lilrb3、Nos2 （iNOS）、Rab2b、Rsad2和Vpreb1的上调。这就得到了一个完整的列表，分别有38个和37个被CR上调和下调表达的注释基因（补充表3）。RT-PCR分析显示，lps诱导的tnf - α、IL-1 β、IL-6和MOS mrna的表达在CR提取物存在下减弱。结果提示，CR通过下调lps刺激的BV-2细胞中与炎症反应相关的多种基因的表达，具有抗炎作用，可能成为预防或治疗炎症介导的神经退行性疾病的候选药物。(C) 2009 Elsevier B.V.版权所有
It has been reported that components of Cinnamomi Ramulus (CR) demonstrate an anti-inflammatory effect by inhibiting the expression of inducible nitric oxide synthesis (iNOS) and cyclooxygenase-2 (COX-2) and by suppressing nitric oxide (NO) production in the central nervous system (CNS) as well as in the periphery. In this study, microarray analysis was performed to investigate the effect of CR on the gene expression and associated pathways of lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Microglia plays an important role in the processes of several inflammation-mediated neurodegenerative diseases. Activated microglia can produce various pro-inflammatory cytokines, chemokines, and toxic mediators, which may initiate or amplify the inflammatory responses in the CNS. In the present study, the negative control group was cultured in normal medium, the positive control group was activated with 1 mu g/ml of LPS, and the CR group was previously treated with 10 mu g/ml of CR before LPS stimulation. With the cutoff value of 1.5-fold change in the expression, 341 genes including pro-inflammatory cytokines, chemokines, and transcription factors were found to be up-regulated in LPS-stimulated BV-2 cells (Supplemental Table 2). CR reduced the LPS-induced up-regulation of such inflammatory genes as Ccl5, Cd80, Cxcl10, Grin2a, Ifi203, Ifit1, Il1 alpha, 116, Lilrb3, Nos2 (iNOS), Rab2b, Rsad2 and Vpreb1. This resulted in a full list of 38 and 37 annotated genes whose expression is up- and down-regulated by CR respectively (Supplemental Table 3). RT-PCR analysis showed that the expression of LPS-induced TNF-alpha, IL-1 beta, IL-6, and MOS mRNAs were attenuated in the presence of the CR extract. The results imply that CR has the anti-inflammatory effect of down-regulating the expression of various genes related to inflammatory responses in LPS-stimulated BV-2 cells, and that CR could be a candidate for the prevention or therapeutic treatment of inflammation-mediated neurodegenerative diseases. (C) 2009 Elsevier B.V. All rights reserved.