萝卜硫素通过调节巨噬细胞 M1/M2 极化减轻炎症环境中巨噬细胞诱导的 Caco-2 细胞损伤。

Sulforaphane Regulates Macrophage M1/M2 Polarization to Attenuate Macrophage-induced Caco-2 Cell Injury in an Inflammatory Environment.

机构信息

Department of Hematology, The Affiliated Changsha Central Hospital, Hengyang Medical School, University of South China, Changsha, Hunan, China.

Department of Medical Administration, The Affiliated Changsha Central Hospital, Hengyang Medical School, University of South China, Changsha, Hunan, China.

出版信息

Iran J Immunol. 2024 Mar 12;21(1):37-52. doi: 10.22034/iji.2024.98644.2580.

DOI:10.22034/iji.2024.98644.2580

PMID:38314669

Abstract

BACKGROUND

The imbalance between M1 and M2 macrophage activation is closely associated with the pathogenesis of inflammatory bowel diseases (IBDs). Sulforaphane (SFN) plays an important role in the treatment of inflammatory diseases.

OBJECTIVE

To investigate the effect of SFN on macrophage polarization and its underlying regulatory mechanism.

METHODS

Mouse bone marrow-derived macrophages (BMDMs) were treated with SFN and an Nrf2 inhibitor, Brusatol. M1 macrophages were induced by LPS and IFN-γ stimulation, whereas M2 macrophages were induced by stimulation with IL-4 and IL-13. LPS-stimulated BMDMs were co-cultured with Caco-2 cells. Flow cytometry, qRT-PCR, and Western blot were performed to assess macrophage polarization. Cell function was assessed using CCK8 assay, transepithelial electrical resistance (TEER) assay, and biochemical analysis.

RESULTS

Higher concentrations of SFN resulted in better intervention effects, with an optimal concentration of 10 μM. SFN decreased the levels of IL-12, IL-6, and TNF-α, as well as the percentages of CD16/32 in M1 BMDMs. At the same time, SFN increased the levels of YM1, Fizz1, and Arg1 as well as the percentages of CD206+ cells in M2 BMDMs. In addition, SFN enhanced the accumulation of Nrf2, NQO1, and HO-1 in M1 BMDMs, and the downregulation of Nrf2 reversed the regulatory effect of SFN on M1/M2 macrophages. LPS-stimulated BMDMs induced Caco-2 cell damage, which was partially alleviated by SFN.

CONCLUSION

Our findings indicate that SFN may act as an Nrf2 agonist to regulate macrophage polarization from M1 to M2. Furthermore, SFN may represent a potential protective ingredient against IBD.

摘要

背景

M1 和 M2 巨噬细胞激活失衡与炎症性肠病（IBD）的发病机制密切相关。萝卜硫素（SFN）在炎症性疾病的治疗中起着重要作用。

目的

探讨 SFN 对巨噬细胞极化的影响及其潜在的调节机制。

方法

用 SFN 和 Nrf2 抑制剂 Brusatol 处理小鼠骨髓来源的巨噬细胞（BMDMs）。用 LPS 和 IFN-γ 刺激诱导 M1 巨噬细胞，用 IL-4 和 IL-13 刺激诱导 M2 巨噬细胞。将 LPS 刺激的 BMDMs 与 Caco-2 细胞共培养。通过流式细胞术、qRT-PCR 和 Western blot 评估巨噬细胞极化。通过 CCK8 测定、跨上皮电阻（TEER）测定和生化分析评估细胞功能。

结果

较高浓度的 SFN 干预效果更好，最佳浓度为 10 μM。SFN 降低了 M1 BMDMs 中 IL-12、IL-6 和 TNF-α的水平，以及 CD16/32 的百分比。同时，SFN 增加了 M2 BMDMs 中 YM1、Fizz1 和 Arg1 的水平以及 CD206+细胞的百分比。此外，SFN 增强了 M1 BMDMs 中 Nrf2、NQO1 和 HO-1 的积累，而 Nrf2 的下调逆转了 SFN 对 M1/M2 巨噬细胞的调节作用。LPS 刺激的 BMDMs 诱导 Caco-2 细胞损伤，SFN 部分缓解了这种损伤。