Park Hae Jeong, Kim Hak Jae, Ra Jehyun, Hong Seung-Jae, Baik Hyung Hwan, Park Hun-Kuk, Yim Sung Vin, Nah Seong-Su, Cho Jeong Je, Chung Joo-Ho
Kohwang Medical Research Institute, School of Medicine, Kyung Hee University, Seoul, Korea.
J Pineal Res. 2007 Sep;43(2):121-9. doi: 10.1111/j.1600-079X.2007.00452.x.
Melatonin possesses a number of important biologic activities including oncostatic, anti-oxidant, and immunostimulatory actions. This study was designed to assess the effects of melatonin on inflammation-related gene expression in lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs), using CombiMatrix 2K Human Inflammation chip. After pretreatment with melatonin (100 microm) for 4 hr, cells were incubated with LPS (1 microg/mL) for 24 hr. We compared gene expression profiles between LPS-treated, melatonin-treated, LSP/melatonin-treated, and control groups. LPS induced the upregulation of 95 genes, compared with controls. Melatonin pretreatment in LPS-stimulated PBMCs suppressed the expression of 23 genes more than twofold. Interestingly, melatonin showed a suppressive effect on the expression of CC chemokine subfamily genes, including CCL2/MCP1, CCL3/MIP1 alpha, CCL4/MIP1 beta, CCL5/RANTES, CCL8/MCP2, CCL20/MDC, and CCL22/MIP3 alpha, in LPS-stimulated PBMCs. This result was confirmed by reverse transcriptase polymerase chain reaction. Among the CC chemokine subfamily genes, particularly, the expression of CCL2 and CCL5 was markedly downregulated by melatonin in LPS-stimulated PBMCs. The secretion levels of CCL2 and CCL5 were measured using enzyme-linked immunosorbent assay. Stimulation of PBMCs by LPS induced the secretion of CCL2 (2334.3 +/- 161.4 pg/mL, mean +/- S.E.M.), whereas melatonin pretreatment (153.0 +/- 3.8 pg/mL) inhibited the LPS-induced secretion of CCL2. Melatonin pretreatment (2696.2 +/- 385.3 pg/mL) also inhibited the LPS-induced secretion of CCL5 (4679.6 +/- 107.5 pg/mL). Taken together, these results suggest that melatonin may have a suppressive effect on LPS-induced expression of CC chemokine genes, especially CCL2 and CCL5, which may explain its beneficial effects in the treatment of various inflammatory conditions.
褪黑素具有多种重要的生物学活性，包括抑癌、抗氧化和免疫刺激作用。本研究旨在使用CombiMatrix 2K人类炎症芯片评估褪黑素对脂多糖（LPS）刺激的人外周血单个核细胞（PBMC）中炎症相关基因表达的影响。用褪黑素（100微摩尔）预处理4小时后，将细胞与LPS（1微克/毫升）孵育24小时。我们比较了LPS处理组、褪黑素处理组、LSP/褪黑素处理组和对照组之间的基因表达谱。与对照组相比，LPS诱导了95个基因的上调。在LPS刺激的PBMC中，褪黑素预处理抑制了23个基因的表达，抑制程度超过两倍。有趣的是，褪黑素对LPS刺激的PBMC中CC趋化因子亚家族基因的表达具有抑制作用，这些基因包括CCL2/MCP1、CCL3/MIP1α、CCL4/MIP1β、CCL5/RANTES、CCL8/MCP2、CCL20/MDC和CCL22/MIP3α。这一结果通过逆转录聚合酶链反应得到证实。在CC趋化因子亚家族基因中，特别是CCL2和CCL5的表达在LPS刺激的PBMC中被褪黑素显著下调。使用酶联免疫吸附测定法测量CCL2和CCL5的分泌水平。LPS刺激PBMC诱导了CCL2的分泌（2334.3±161.4皮克/毫升，平均值±标准误），而褪黑素预处理（153.0±3.8皮克/毫升）抑制了LPS诱导的CCL2分泌。褪黑素预处理（2696.2±385.3皮克/毫升）也抑制了LPS诱导的CCL5分泌（4679.6±107.5皮克/毫升）。综上所述，这些结果表明褪黑素可能对LPS诱导的CC趋化因子基因表达具有抑制作用，尤其是CCL2和CCL5，这可能解释了其在治疗各种炎症性疾病中的有益作用。
