| 引用本文: |
何超平,陈煜,彭莎,石哲,李亚梅,廖端芳.乙酰水杨酸姜黄素酯通过TLR4/NF-κB通路拮抗LPS诱导的BV2小胶质细胞炎症损伤[J].湖南中医药大学学报,2022,42(7):1070-1075[点击复制] |
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| 乙酰水杨酸姜黄素酯通过TLR4/NF-κB通路拮抗LPS诱导的BV2小胶质细胞炎症损伤 |
| 何超平,陈煜,彭莎,石哲,李亚梅,廖端芳 |
| (湖南中医药大学药学院湘产大宗药材品质评价湖南省重点实验室, 湖南 长沙 410208) |
| 摘要: |
| 目的 研究乙酰水杨酸姜黄素酯（curcumin acetylsalicylate,CA）对脂多糖（lipopolysaccharide,LPS）诱导的BV2小胶质细胞炎症的改善作用及机制。方法 应用Discovery Studio分子模拟软件将CA与核因子-κB（nuclear factor-κB,NF-κB）、胰岛素诱导基因2（nsulin inducible gene 2,Insig2）、Wnt信号蛋白（Wnt）、尼曼匹克C1型L1（Niemann-Pick C1 like 1,NPC1-L1）、Toll样受体2（Toll-like receptors 2,TLR2）和Toll样受体4（Toll-like receptors 4，TLR4）蛋白进行分子对接，筛选出结合紧密的蛋白。将BV2小胶质细胞分为对照组、模型组、CA干预组。对照组不做处理；模型组加入10 mg/L的LPS作用24 h；CA干预组加入20、40、60 μmol/L的CA作用2 h后，加入10 mg/L LPS作用24 h。CCK-8法检测CA对BV2小胶质细胞增殖的影响；ELISA法检测细胞TNF-α、IL-1β；Griess法检测细胞分泌NO的水平；Western blot法检测iNOS、NF-κB p65、p-NF-κB p65、IκB-α、p-IκB-α、TLR4和MYD88蛋白的表达。结果 与对照组比较，在0~80 μmol/L浓度范围内，CA干预组细胞存活率无显著性差异（P＞0.05）。与对照组比较，模型组细胞TNF-α、IL-1β、NO水平明显升高（P<0.05）；与模型组比较，CA干预组细胞TNF-α、IL-1β、NO水平明显下降（P<0.05）。与对照组比较，模型组的p-NF-κB水平升高（P<0.05）；与模型组比较，CA干预组降低p-NF-κB水平（P<0.05）。与对照组比较，模型组的TLR4、MYD88的蛋白表达水平明显升高（P<0.05）；与模型组比较，CA干预组的TLR4、MYD88的蛋白表达水平明显降低（P<0.05）。结论 CA能有效抑制LPS诱导的BV2小胶质细胞炎症反应，其作用可能与其抑制NF-κB/TLR4信号通路有关。 |
| 关键词: 乙酰水杨酸姜黄素酯 BV2小胶质细胞 神经炎症 核因子-κB Toll样受体4 |
| DOI：10.3969/j.issn.1674-070X.2022.07.003 |
| 投稿时间：2022-01-12 |
| 基金项目:湖南省科技创新计划项目（2021RC4064）；湖南省重点研发计划项目（2022SK2011）；湖南省教育厅重点项目（20A379）。 |
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| Curcumin acetylsalicylate antagonizes LPS-induced inflammatory injury of BV2 microglia through TLR4/NF-κB pathway |
| HE Chaoping,CHEN Yu,PENG Sha,SHI Zhe,LI Yamei,LIAO Duanfang |
| (School of Pharmaceutical Science, Hunan University of Chinese Medicine, Key Laboratory for Quality Evaluation of Bulk Herbs of Hunan Province, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To research the effect and mechanism of curcumin acetylsalicylate (CA) on lipopolysaccharide (LPS)-induced BV2 microglial inflammation. Methods Discovery Studio molecular modeling package was used to docking CA with nuclear factor-κB (NF-κB), Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) proteins to screen out the tightly bound proteins. BV2 microglia cells were divided into control group, model group and CA administration group. The control group was not treated; the model group was added with 10 mg/L LPS for 24 h; the CA administration group was treated with 10 mg/L CA for 24 h, and then added 20, 40, 60 μmol/L LPS for 24 h. The effect of CA on the proliferation of BV2 cells was detected by CCK-8. ELISA method was used to detect the cellular TNF-α and IL-1β; the level of NO secreted by cells was detected by Griess method; the expression levels of iNOS, NF-κB p65, p-NF-κB p65, IκB-α, p-IκB-α, TLR4 and MYD88 protein were detected by Western blot. Results Compared with the control group, in the concentration range of 0-80 μmol/L, the cell viability of the CA administration group had no significant difference (P>0.05); compared with the control group, the levels of TNF-α, IL-1β and NO in the model group were significantly increased (P<0.05); compared with the model group, the levels of TNF-α, IL-1β and NO in the CA administration group were significantly decreased (P<0.05). Compared with the control group, the level of p-NF-κB in the model group was increased (P<0.05); compared with the model group, the level of p-NF-κB in the CA administration group was reduced (P<0.05). Compared with the control group, the protein expression levels of TLR4 and MYD88 in the model group were significantly increased (P<0.05); compared with the model group, the protein expression levels of TLR4 and MYD88 in the CA administration group were significantly decreased (P<0.05). Conclusion CA can effectively inhibit the inflammatory response of BV2 microglial cells induced by LPS, and its effect may be related to its inhibition of NF-κB/TLR4 signaling pathway. |
| Key words: curcumin acetylsalicylate BV2 microglia neuroin flammation nuclear factor-κB Toll-like receptor 4 |
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