上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (5): 446-.doi: 10.3969/j.issn.1674-8115.2019.05.002

• 论著·基础研究 • 上一篇    下一篇

金雀异黄素对脂多糖诱导小胶质细胞炎症反应的抑制作用

王红梅 ，付剑亮，张婷，陈静炯，赵玉武   

  1. 上海交通大学附属第六人民医院神经内科，上海 200233
  • 出版日期:2019-05-28 发布日期:2019-07-26
  • 通讯作者: 王红梅，电子信箱：dd-tt12345@163.com。赵玉武，电子信箱：zhaoyuwu2005@126.com。为共同通信作者。
  • 作者简介:王红梅（1975—），女，副主任医师，博士；电子信箱： dd-tt12345@163.com。
  • 基金资助:
    国家自然科学基金（81672243，31771185，81871103，81870952）

Inhibition of genistein against LPS-induced proinflammatory response in microglia

WANG Hong-mei, FU Jian-liang, ZHANG Ting, CHEN Jing-jiong, ZHAO Yu-wu   

  1. Department of Neurology, Shanghai Sixth Peoples Hospital, Shanghai Jiao Tong University, Shanghai 200233, China
  • Online:2019-05-28 Published:2019-07-26
  • Supported by:
    National Natural Science Foundation of China, 81672243, 31771185, 81871103, 81870952

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摘要： 目的 ·探讨金雀异黄素（ genistein，Gen）对脂多糖（ lipopolysaccharide，LPS）诱导小胶质细胞炎症反应的抑制作用。方法 ·取新生 1 d的 C57BL/6J小鼠分离原代小胶质细胞进行培养，并分为 LPS组、Gen+LPS组、对照组。 LPS组给予 LPS（1 μg/mL）处理 24 h，Gen+LPS组给予 Gen（10 μmol/L）预处理 0.5 h后加入 LPS（1 μg/mL）处理 24 h，对照组不做处理。采用 Western blotting分析 CD11b蛋白的表达，采用 MitoSOXTM红色试剂和 CM-H2DCFDA分别观察线粒体活性氧（ reactive oxygen species，ROS）和细胞内 ROS水平，通过免疫荧光染色、比色法和 ELISA法分别检测 NLRP3（NLR family pyrin domain containing 3）炎症小体、胱天蛋白酶 1（caspase-1）和白细胞介素 1β（interleukin-1β，IL-1β）的变化。结果 ·与对照组比较， LPS组小胶质细胞 CD11b蛋白水平，及线粒体、细胞内 ROS水平均显著升高（均 P<0.05），而 Gen+LPS组明显低于 LPS组（均 P<0.05）。此外， LPS组小胶质细胞 NLRP3炎症小体免疫荧光强度、 caspase-1活性和 IL-1β分泌显著高于对照组（均 P<0.05）；与 LPS组相比， Gen+LPS组均明显下降（均 P<0.05）。结论 · Gen可抑制 LPS诱导的小胶质细胞线粒体 ROS生成、NLRP3炎症小体活化和炎症因子 IL-1β分泌等炎症反应。

关键词: 金雀异黄素, 小胶质细胞, 脂多糖, 炎症, NLRP3炎症小体, 活性氧

Abstract:

Objective · To investigate the effect of genistein (Gen) on lipopolysaccharide (LPS)-induced proinflammatory response in microglia. Methods · Primary microglia were isolated C57BL/6J mice 1 d after birth, which were divided into 3 groups, i.e. control group, LPS group, and Gen+LPS group. Microglia in LPS group and Gen+LPS group were incubated with LPS (1 μg/mL) for 24 h, and the cells in Gen+LPS group were also pretreated with Gen (10 μmol/L) for 0.5 h. The of CD11b was measuredWestern blotting analysis. Besides, mitochondrial and intracellular reactive oxygen species (ROS) levels were monitoredMitoSOXTM Red and CM-H2DCFDA staining, respectively. NLRP3 (NLR family pyrin domain containing 3) inflammasom was detectedimmunofluorescence. Caspase-1 activity and interleukin-1β (IL-1β) levels were evaluatedcolorimetric assay kit and ELISA kit, respectively. Results · Compared with control group, LPS increased CD11b protein , and mitochondrial and intracellular ROS levels in microglia (P<0.05). And Gen obviously reversed LPS-induced mitochondrial and intracellular ROS accumulation as well as reduced CD11b (P<0.05). In addition, LPS enhanced fluorescence intensity of NLRP3 inflammasome, caspase-1 activity and IL-1β secretion in microglia when compared with control group (P<0.05), while Gen significantly attenuated these effects (P<0.05). Conclusion · Gen can inhibit LPS-induced proinflammatory response including mitochondrial ROS accumulation, activation of NLRP3 inflammasome, and IL-1β secretion in microglia.

Key words: genistein, microglia, lipopolysaccharide (LPS), inflammation, NLRP3 inflammasome, reactive oxygen species (ROS)

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