Shiang Jeng-Chuan, Jan Ren-Long, Tsai Ming-Kai, Hsieh Chong-Chao, Kuo Hsuan-Fu, Kuo Chang-Hung, Yang San-Nan, Huang Ming-Yii, Chen Li-Chen, Hung Chih-Hsing
Graduate Institute of Medicine, Kaohsiung Medical University Hospital; Department of Pediatrics, Faculty of Medicine; Department of Pediatrics, Faculty of Pediatrics, College of Medicine, Kaohsiung Medical University; Department of Pediatrics, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung, Taiwan.
Indian J Med Res. 2014 Jul;140(1):109-15.
BACKGROUND & OBJECTIVES: Selective cyclooxygenase-2 (COX-2) inhibitor is a form of thnon steroidal anti-inflammatory drug (NSAID) and is commonly used in autoimmune and rheumatic diseases to control inflammation and alleviate pain. Tumour necrosis factor-alpha (TNF-α) production and an imbalance of T helper 1 (Th1)/Th2 contribute to the pathogenesis of autoimmune and also anti-tumour activity. Dipyrone is a NSAID used to treat pain worldwide. The celecoxib analogue, 2,5-dimethylcelecoxib (DMC), lacks COX-2 inhibitory activity but exhibits anti-tumour properties. However, the effects and the mechanisms of dipyrone and 2,5-dimethylcelecoxib on tumour necrosis factor (TNF)-α and Th1- and Th2-related chemokines in monocytes remain poorly defined. This study was carried out to investigate the effects of dipyrone and 2,5-dimethylcelecoxib on the expression of Th1 (IP-10) and Th2 (I-309 and MDC) and TNF-α in human monocytes and the associated intracellular mechanism.
THP-1 cells and peripheral blood mononuclear cells (PBMCs) were pre-treated with dipyrone (10(-9)-10(-4) M) and 2,5-dimethylcelecoxib (10(-9)-10(-5) M) 2 h before lipopolysaccharide (LPS) stimulation. Cell supernatant was collected 24 h after LPS stimulation. TNF-α, I-309, MDC and IP-10 concentrations of cell supernatants were determined using ELISA. Intracellular signaling was evaluated by w0 estern blot.
Dipyrone and 2,5-dimethylcelecoxib downregulated LPS-induced Th2-related chemokine I-309 and macrophage derived chemokine (MDC) production. Only high dose of 2,5-dimethylcelecoxib (10(-5) M), but not dipyrone downregulated LPS-induced IP-10. Only very high dose of 2,5-dimethylcelecoxib had effect on LPS-induced TNF-α expression in PBMCs. Dipyrone and 2,5-dimethylcelecoxib suppressed LPS-induced p65 and JNK MAPK (C-Jun N-terminal kinase mitogen activated protein kinase). expression.
INTERPRETATION & CONCLUSIONS: Dipyrone and 2,5-dimethylcelecoxib downregulated LPS-induced Th2-related chemokine I-309 and MDC in THP-1 cells. The suppressive effect on Th2-related chemokine I-309 and MDC may involve the downregulation of LPS-induced JNK and p65 expression.
选择性环氧化酶-2（COX-2）抑制剂是一种非甾体抗炎药（NSAID），常用于自身免疫性疾病和风湿性疾病以控制炎症和缓解疼痛。肿瘤坏死因子-α（TNF-α）的产生以及辅助性T细胞1（Th1）/辅助性T细胞2（Th2）失衡参与自身免疫性疾病的发病机制，同时也具有抗肿瘤活性。安乃近是一种在全球范围内用于治疗疼痛的NSAID。塞来昔布类似物2,5-二甲基塞来昔布（DMC）缺乏COX-2抑制活性，但具有抗肿瘤特性。然而，安乃近和2,5-二甲基塞来昔布对单核细胞中肿瘤坏死因子（TNF）-α以及Th1和Th2相关趋化因子的影响及机制仍不清楚。本研究旨在探讨安乃近和2,5-二甲基塞来昔布对人单核细胞中Th1（IP-10）、Th2（I-309和MDC）以及TNF-α表达的影响及其相关的细胞内机制。
在脂多糖（LPS）刺激前2小时，用安乃近（10⁻⁹ - 10⁻⁴ M）和2,5-二甲基塞来昔布（10⁻⁹ - 10⁻⁵ M）预处理THP-1细胞和外周血单个核细胞（PBMC）。LPS刺激24小时后收集细胞上清液。使用酶联免疫吸附测定（ELISA）法测定细胞上清液中TNF-α、I-309、MDC和IP-10的浓度。通过蛋白质免疫印迹法评估细胞内信号传导。
安乃近和2,5-二甲基塞来昔布下调LPS诱导的Th2相关趋化因子I-309和巨噬细胞衍生趋化因子（MDC）的产生。仅高剂量的2,5-二甲基塞来昔布（10⁻⁵ M）可下调LPS诱导的IP-10，而安乃近无此作用。仅极高剂量的2,5-二甲基塞来昔布对PBMC中LPS诱导的TNF-α表达有影响。安乃近和2,5-二甲基塞来昔布抑制LPS诱导的p65和JNK丝裂原活化蛋白激酶（C-Jun N端激酶丝裂原活化蛋白激酶）的表达。
安乃近和2,5-二甲基塞来昔布下调LPS诱导的THP-1细胞中Th2相关趋化因子I-309和MDC的表达。对Th2相关趋化因子I-309和MDC的抑制作用可能涉及下调LPS诱导的JNK和p65的表达。
