姜辣素通过抑制 NF-κB 信号通路对脂多糖刺激的 RAW 264.7 细胞的抗炎作用。
Anti-Inflammatory Effects of Gingerol on Lipopolysaccharide-Stimulated RAW 264.7 Cells by Inhibiting NF-κB Signaling Pathway.
机构信息
Faculty of Food Science and Technology, Agricultural University of Hebei, Baoding, 071001, People's Republic of China.
Hebei Research Center of Primary Products Processing Technology, Baoding, 071001, People's Republic of China.
出版信息
Inflammation. 2018 Jun;41(3):835-845. doi: 10.1007/s10753-018-0737-3.
Gingerol was the main functional substance of Zingiberaceous plant which has been known as traditional medicine for thousands of years. The purpose of this experiment was to explore anti-inflammatory effects of gingerol and study the possible mechanism in lipopolysaccharide (LPS)-stimulated RAW246.7 cells. The cells were treated with 10 μg/mL LPS and 300, 200, 100, and 50 μg/mL gingerol for 24 h. The cytotoxicity of gingerol was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetra-zoliumbromide (MTT) method. Nitric oxide (NO) production was observed using Griess assays. Prostaglandin E (PGE) and pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 have been analyzed by ELISA. Real-time PCR was used to detect the mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), IL-6, and IL-1β in LPS-induced RAW246.7 cells. Nuclear transcription factor kappa-B (NF-κB) signaling pathway-related proteins have been assessed by western blot assays. The determination of MTT showed that cell viability was not significantly affected by up to 300 μg/mL gingerol. Compared with LPS group, 50, 100, 200, and 300 μg/mL gingerol can inhibit the production of NO and the inhibitory rate was 10.4, 29.1, 58.9, and 62.4%, respectively. The results indicated gingerol existed anti-inflammatory effect. In addition, gingerol also observably inhibited LPS-induced TNF-α, IL-1β, IL-6, and PGE (p < 0.01) expression and secretion in a dose-dependent manner. At the genetic level, after the intervention of gingerol, mRNA transcriptions of iNOS, COX-2, IL-6, and IL-1β were all decreased. The protein expressions of iNOS, NF-κB, p-p65, and p-IκB were significantly increased in LPS-induced cells, while these changes were reversed by the treatment with gingerol. This study suggested that gingerol exerts its anti-inflammatory activities in LPS-induced macrophages which can inhibit the production of inflammatory cytokines by targeting the NF-κB signaling pathway.
姜辣素是姜科植物的主要功能物质，作为传统药物已有数千年的历史。本实验旨在探讨姜辣素的抗炎作用，并研究其在脂多糖（LPS）刺激的 RAW246.7 细胞中的可能机制。用 10μg/mL LPS 和 300、200、100 和 50μg/mL 姜辣素处理细胞 24 小时。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物（MTT）法测定姜辣素的细胞毒性。用 Griess 法观察一氧化氮（NO）的产生。通过 ELISA 分析前列腺素 E (PGE)和促炎细胞因子肿瘤坏死因子-α (TNF-α)、白细胞介素 (IL)-1β 和 IL-6。实时 PCR 用于检测 LPS 诱导的 RAW246.7 细胞中诱导型一氧化氮合酶 (iNOS)、环氧化酶-2 (COX-2)、IL-6 和 IL-1β 的 mRNA 表达。用 Western blot 法测定核转录因子 κB（NF-κB）信号通路相关蛋白。MTT 的测定表明，高达 300μg/mL 的姜辣素对细胞活力没有显著影响。与 LPS 组相比，50、100、200 和 300μg/mL 姜辣素可抑制 NO 的产生，抑制率分别为 10.4%、29.1%、58.9%和 62.4%。结果表明姜辣素有抗炎作用。此外，姜辣素还能显著抑制 LPS 诱导的 TNF-α、IL-1β、IL-6 和 PGE 的表达和分泌（p<0.01），呈剂量依赖性。在遗传水平上，姜辣素干预后，iNOS、COX-2、IL-6 和 IL-1β 的 mRNA 转录均降低。LPS 诱导的细胞中 iNOS、NF-κB、p-p65 和 p-IκB 的蛋白表达均显著增加，而用姜辣素处理可逆转这些变化。本研究表明，姜辣素通过靶向 NF-κB 信号通路抑制炎症细胞因子的产生，发挥 LPS 诱导的巨噬细胞的抗炎活性。
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