[1]齐世美,李强,姜琦,等.白杨素通过JAK-STATs 信号通路抑制内毒素诱导的巨噬细胞炎症反应[J].南方医科大学学报,2018,(03):243.
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白杨素通过JAK-STATs 信号通路抑制内毒素诱导的巨噬细胞炎症反应()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2018年03期
页码:
243
栏目:
出版日期:
2018-04-03

文章信息/Info

Title:
Chrysin inhibits lipopolysaccharide-induced inflammatory responses of macrophages via JAK-STATs signaling pathway
作者:
齐世美李强姜琦戚之琳章尧
关键词:
白杨素活性氧簇RAW264.7细胞JAK-STATs信号通路
Keywords:
chrysin reactive oxygen species RAW264.7 cells JAK-STATs
摘要:
目的探讨白杨素对LPS诱导的RAW264.7细胞炎症的调控机制。方法分别用不同浓度(0、5、10、20、40、60、80、100、150、 200 μg/mL)白杨素作用RAW264.7 细胞24 h 后,采用CCK-8 检测细胞活力值;分别用(10、30、60 μg/mL)白杨素预处理 RAW264.7细胞2 h后,用LPS刺激RAW264.7细胞18 h后,采用ELISA法检测炎症因子TNF-α、IL-6和MCP-1的释放水平;分 别用(10、30、60 μg/mL)白杨素预处理RAW264.7 细胞2 h 后,用LPS刺激RAW264.7 细胞4 h 后,运用Western blotting 法检测 JAK-1、JAK-2、STAT-1、STAT-3 的磷酸化水平;分别用(10、30、60 μg/mL)白杨素预处理RAW264.7 细胞2 h 后,用LPS 刺激 RAW264.7 细胞15 min 后,运用CM-H2DCFDA荧光探针检测RAW264.7 细胞内活性氧簇水平;运用ROS清除剂NAC处理 RAW264.7细胞,检测ROS对LPS诱导RAW264.7细胞JAK-STATs信号和炎症反应的影响;运用激光共聚焦显微镜观察转录因 子STAT-1和STAT-3核转位情况。结果在白杨素浓度低于60 μg/mL时,对RAW264.7细胞活力没有显著影响,因此我们选择 10、30、60 μg/mL白杨素作为抑制炎症作用的低、中、高剂量组;白杨素剂量依赖性地抑制LPS诱导的炎症蛋白iNOS的表达;白 杨素剂量依赖性的下调LPS诱导的RAW264.7细胞中促炎因子TNF-α、IL-6和MCP-1的释放(P<0.01);白杨素抑制RAW264.7 细胞中JAK-STATs信号活化并抑制STAT-1和STAT-3核转位;白杨素抑制RAW264.7细胞内ROS的产生;ROS作为上游信号介 导JAK-STATs信号通路的活化。结论白杨素能有效阻断ROS介导的JAK-STATs信号活化,从而抑制LPS诱导的RAW264.7 细胞炎症反应。
Abstract:
Objective To investigate the mechanism of chrysin in regulating lipopolysaccharide (LPS)-induced inflammation in RAW264.7 cells. Methods RAW264.7 cells were treated with different concentrations (0, 5, 10, 20, 40, 60, 80, 100, 150, and 200 μg/mL) of chrysin for 24 h, and the cell viability was measured using CCK-8. RAW264.7 cells were pre-treated with 10, 30, or 60 μg/mL chrysin for 2 h before stimulation with LPS for different times. The levels of TNF-α, IL-6 and MCP-1 were detected by ELISA, and Western blotting was used to detect the phosphorylation of JAK- 1, JAK-2, STAT-1 and STAT-3. The level of reactive oxygen species in RAW264.7 cells was detected by CM-H2DCFDA fluorescence probe. The effect of ROS on LPS-induced JAK-STATs signal and the inflammatory response of RAW264.7 cells was detected by ROS scavenger NAC. The transcription factors STAT-1 and STAT-3 nuclear translocation were observed by laser confocal microscopy. Results Chrysin below 60 μg/mL did not significantly affect the viability of RAW264.7 cells. At 10, 30, and 60 μg/mL, chrysin dose-dependently inhibited the expression of iNOS induced by LPS. Chrysin treatment also inhibited LPS-induced phosphorylation of JAK-STATs, nuclear translocation of STAT1 and STAT3, release of TNF-α, IL-6 and MCP-1, and the production of ROS in RAW264.7 cells; ROS acted as an upstream signal to mediate the activation of JAKSTATs signaling pathway. Conclusion Chrysin blocks the activity of JAK-STATs mediated by ROS to inhibit LPSinduced inflammatory response in RAW264.7 cells.

相似文献/References:

[1]韦小彤,彭文蕊,姜琦,等.白杨素通过调控MAPKs信号通路促进SMMC-7721细胞凋亡[J].南方医科大学学报,2018,(10):1187.[doi:10.12122/j.issn.1673-4254.2018.10.06]

更新日期/Last Update: 1900-01-01