天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (11): 1857-1863.doi: 10.16333/j.1001-6880.2023.11.003

• 研究论文 • 上一篇    下一篇

白术内酯Ⅱ逆转M2型巨噬细胞极化调控PDL1表达抑制A549细胞迁移

蒋宗蓥,赵云辉,张云亭,郑   一,王建光,刘羽茜,王艳杰*   

  1. 辽宁中医药大学中西医结合学院,沈阳 110847
  • 出版日期:2023-11-28 发布日期:2023-11-25
  • 基金资助:
    国家自然科学基金青年基金(81202789);辽宁省自然科学基金指导计划(2019-ZD-0950);辽宁省教育厅一般项目(LJKM20221311)

AtractylenolideⅡ inhibits A549 cell migration by reversing M2 type macrophage polarization and regulating PDL1 expression

JIANG Zong-ying,ZHAO Yun-hui,ZHANG Yun-ting,ZHENG Yi,WANG Jian-guang,LIU Yu-xi,WANG Yan-jie*   

  1. College of Integrated Traditional Chinese and Western Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China
  • Online:2023-11-28 Published:2023-11-25

摘要:

探究白术内酯Ⅱ(atractylenolideⅡ,AT-Ⅱ)对M2型巨噬细胞极化与A549细胞迁移能力的影响,并分析其潜在机制。利用PMA将THP-1细胞诱导成M0巨噬细胞,然后加入IL-4和IL-13继续诱导为M2型巨噬细胞。利用Transwell小室将M2型巨噬细胞与A549细胞共培养,分别给予0、2.5、5 μmol/L的AT-Ⅱ进行作用。采用MTT实验测定共培养条件下AT-Ⅱ对A549细胞活力的影响;采用Real-time PCR检测M2型巨噬细胞Arg-1的 mRNA表达水平;采用ELISA检测共培养体系中TNF-α、IL-1β的含量;采用WB检测A549细胞TLR4、p-p65、NF-κB(p65)、PDL1的蛋白表达水平;采用划痕实验检测A549细胞的迁移能力。结果显示,共培养条件下,与对照组相比,实验组(2.5、5 μmol/L)A549细胞活力降低(2.5 μmol/L组P>0.05、5 μmol/L组P<0.01);M2型巨噬细胞特异性基因Arg-1的mRNA表达水平显著降低(P<0.01);M1型巨噬细胞相关炎性因子TNF-α、IL-1β含量增多但无显著差异(P>0.05);A549细胞TLR4/NF-κB信号通路相关蛋白TLR4、p-p65、NF-κB(p65)蛋白表达水平降低(P<0.01);A549细胞PDL1蛋白表达水平显著降低(P<0.01),A549细胞迁移能力下降。以上结果表明,AT-Ⅱ可以逆转M2型巨噬细胞极化,抑制A549细胞TLR4/NF-κB信号通路降低PDL1的表达水平,从而抑制肺癌细胞的迁移。

关键词: 白术内酯Ⅱ, A549细胞, M2型巨噬细胞, PDL1蛋白

Abstract:

To explore the effect of atractylenolide Ⅱ(AT-Ⅱ) on the polarization of M2 macrophages and the migration of A549 cells,and to analyze its potential mechanism.THP-1 cells were induced into M0 macrophages using PMA,and then IL-4 and IL-13 were added to continue inducing M0 macrophages into M2 macrophages.M2 macrophages were co-cultured with A549 cells in Transwell chambers and treated with 0,2.5,5 μmol/L AT-Ⅱ.MTT assay was used to determine the effect of AT-Ⅱ on the viability of A549 cells under co-culture condition;Real-time PCR was used to detect the mRNA expression level of Arg-1;ELISA was used to detect TNF-α,IL-1β content in co-culture system;WB was used to detect the protein expression levels of TLR4,p-p65,NF-κB (p65),PDL1;Scratch experiment was used to detect the migration ability of A549 cells.The results showed that under co-culture condition,compared with the control group,the activity of A549 cells in the experimental group decreased,2.5 μmol/L group (P>0.05),5 μmol/L group (P<0.01);The mRNA expression level of M2 macrophage specific gene Arg-1 was significantly reduced (P<0.01);The content of M1 macrophage associated inflammatory factor TNF-α,IL-1β increased,but did not constitute a significant difference (P>0.05);The expression levels of TLR4,p-p65 and NF-κB (p65) proteins related to the TLR4/NF-κB signaling pathway decreased in A549 cells (P<0.01);The expression of PDL1 protein in A549 cells significantly decreased (P<0.01),and the ability of A549 cells migration decreased.These results indicate that AT-Ⅱ can reverse the polarization of M2 macrophages,inhibit NF-κB signaling pathway in A549 cells and reduce the expression level of PDL1,thus inhibiting the migration of lung cancer cells.

Key words: atractylenolide Ⅱ, A549 cells, M2 macrophages, PDL1 protein

中图分类号:  R34