基于NF-κB通路探究重楼皂苷Ⅶ的抗炎作用机制
Anti-Inflammatory Mechanism of Paris Saponin Ⅶ via NF-κB Signaling Pathway
晋兰 1吴琼 1梁金美琪 1林迪斯 2郗雪艳 1郭阳1
作者信息
- 1. 湖北医药学院基础医学院,湖北 十堰 442000
- 2. 湖北医药学院附属国药东风总医院病理科,湖北 十堰 442000
- 折叠
摘要
目的:探讨重楼皂苷Ⅶ(PSⅦ)的抗炎作用及机制.方法:利用MACS法从C57BL/6 小鼠脾脏中分离初始T淋巴细胞.PSⅦ处理后,通过流式细胞术、RT-qPCR、Western blot检测PSⅦ对IL-6 表达水平的影响;利用流式细胞术、Cell TraceTM Violet试剂盒检测PSⅦ对CD4+T细胞活化及增殖分化的影响;利用Western blot检测PSⅦ对IκBα、STAT3 及其磷酸化蛋白表达的影响;RNA-seq检测PSⅦ处理后的差异基因.结果:PSⅦ显著抑制IL-6 的表达.PSⅦ对CD4+T细胞的活化及增殖无明显影响,但对其分化有一定影响.PSⅦ抑制p-IκBα、p-STAT3 的蛋白表达水平,但对IκBα、STAT3 无影响.RNA-seq分析的差异基因有 70 个(27 个上调、43 个下调),从中筛选出 6个并利用RT-qPCR得到验证.结论:PSⅦ可通过STAT3 和NF-κB信号通路抑制IL-6 的表达,从而缓解炎症.
Abstract
Objective To decipher the anti-inflammatory mechanism of Paris saponin Ⅶ(PSⅦ).Methods Naive T lym-phocytes were isolated from the spleen of C57BL/6 mice by magnetic-activated cell sorting.After Paris saponin Ⅶ treat-ment,flow cytometry,RT-qPCR and Western blotting were employed to examine the effects of PSⅦon the expression lev-el of interleukin-6(IL-6).The effects of PSⅦon the activation,proliferation and differentiation of CD4+T cells were de-tected by flow cytometry and Cell TraceTM Violet kit.Western blot was used to determine the expression and phosphoprotein of nuclear factor-kappa beta inhibitor alpha(IκBα)and signal transducer and activator of transcription 3(STAT3)after PSⅦ treatment.The differentially expressed genes(DEGs)after PSⅦ treatment were detected by RNA-seq.Results PSⅦ significantly inhibited the expression of IL-6.PSⅦ had no obviously effect on the activation and proliferation,but can influenced the differentiation of CD4+T cells.PSⅦcan inhibited the phosphoprotein of IκBα and STAT3,but had no effect on the expression of the two proteins.A total of 70 DEGs were detected by RNA-seq,including 27 up-regulated genes and 43 down-regulated genes,from which 6 genes were selected and verified by RT-qPCR.Conclusion PSⅦ can relieve in-flammation by inhibiting IL-6 expression via STAT3 and NF-κB signaling pathways.
关键词
重楼皂苷Ⅶ/NF-κB信号通路/白细胞介素-6/炎性反应Key words
Paris saponin Ⅶ/NF-κB signaling pathway/IL-6/Inflammation引用本文复制引用
基金项目
武当特色中药研究湖北省重点实验室开放课题(WDCM2023020)
湖北省重点研发计划(2022BCE040)
国药东风总医院青年人才项目(2022Q21)
出版年
2024
国家科技期刊平台
NSTL
万方数据