3,3'-二吲哚甲烷，一种天然芳基烃受体激动剂，通过增强“糖酵解-乳酸-STAT3”和 TIP60 信号介导的 Treg 分化来缓解溃疡性结肠炎。
3, 3'-diindolylmethane, a natural aryl hydrocarbon receptor agonist, alleviates ulcerative colitis by enhancing "glycolysis-lactate-STAT3″ and TIP60 signals-mediated Treg differentiation.
机构信息
Department of Pharmacology of Chinese Materia Medica, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, China.
Department of Pharmacology of Chinese Materia Medica, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, China.
出版信息
Mol Immunol. 2023 Nov;163:147-162. doi: 10.1016/j.molimm.2023.09.009. Epub 2023 Oct 2.
BACKGROUND
Aryl hydrocarbon receptor (AhR) plays an important role in the occurrence and development of ulcerative colitis (UC). In this study, the effect and mechanism of 3, 3'-diindolylmethane (DIM), the classical AhR agonist, on UC was investigated from the angle of recovering the balance of Th17/Treg.
METHODS
The in vivo colitis model was established in mice by using dextran sulfate sodium, and CD4 T cells were used to simulate the in vitro differentiation of Treg and Th17 cells. The proportions and related factors of Th17 and Treg cells were measured using flow cytometry, Q-PCR and western blotting. The glycolysis was evaluated by examining the glucose uptake, glucose consumption and lactate production using kits or immunofluorescence. The activation of AhR was detected by western blotting and the XRE-luciferase reporter gene. The co-immunoprecipitation, transfection or other methods were selected to investigate and identify the signaling molecular pathway.
RESULTS
DIM significantly attenuated symptoms of colitis mice by rebuilding the balance of Th17/Treg in anoxic colons. In hypoxia, a more potent promotion of Treg differentiation was showed by DIM relative to normoxia, and siFoxp3 prevented DIM-suppressed Th17 differentiation. DIM repressed the excessive glycolysis in hypoxia evidenced by down-regulated glucose uptake, lactate production, Glut1 and HK2 levels. Interestingly, IL-10, the function-related factor of Treg cells, showed the feedback effect of DIM-suppressed glycolysis. Besides, 2-deoxy-D-glucose, HK2 plasmid and IL-10 antibody prevented increase of DIM on the expression of Foxp3 at the transcriptional level and subsequent Treg differentiation through the lactate-STAT3 pathway, and reasons for the direct improvement of DIM on Foxp3 protein was attributed to promoting the formation of HIF-1α/TIP60 complexes as well as subsequent acetylation and protein stability. Finally, AhR dependence and mechanisms for DIM-improved Treg differentiation in vitro and in vivo were well confirmed by using plasmids or inhibitors.
CONCLUSIONS
DIM enhances activation of AhR and subsequent "glycolysis-lactate-STAT3″ and TIP60 signals-mediated Treg differentiation.
背景
芳香烃受体（AhR）在溃疡性结肠炎（UC）的发生和发展中起着重要作用。在这项研究中，从恢复 Th17/Treg 平衡的角度研究了经典 AhR 激动剂 3,3'-二吲哚甲烷（DIM）对 UC 的作用和机制。
方法
采用葡聚糖硫酸钠建立小鼠结肠炎模型，用 CD4 T 细胞模拟 Treg 和 Th17 细胞的体外分化。采用流式细胞术、Q-PCR 和 Western blot 检测 Th17 和 Treg 细胞的比例及相关因子。用试剂盒或免疫荧光法检测葡萄糖摄取、葡萄糖消耗和乳酸产生来评估糖酵解。用 Western blot 和 XRE-荧光素酶报告基因检测 AhR 的激活。选择共免疫沉淀、转染或其他方法来研究和鉴定信号分子通路。
结果
DIM 通过重建缺氧结肠中的 Th17/Treg 平衡，显著减轻结肠炎小鼠的症状。在缺氧条件下，与常氧相比，DIM 更能促进 Treg 分化，而 siFoxp3 则阻止了 DIM 抑制 Th17 分化。DIM 通过下调葡萄糖摄取、乳酸生成、Glut1 和 HK2 水平来抑制缺氧时的过度糖酵解。有趣的是，IL-10，Treg 细胞的功能相关因子，显示了 DIM 抑制糖酵解的反馈效应。此外，2-脱氧-D-葡萄糖、HK2 质粒和 IL-10 抗体通过乳酸-STAT3 通路阻止了 DIM 对转录水平 Foxp3 表达的增加以及随后的 Treg 分化，并认为 DIM 对 Foxp3 蛋白的直接改善归因于促进 HIF-1α/TIP60 复合物的形成以及随后的乙酰化和蛋白质稳定性。最后，通过使用质粒或抑制剂，在体外和体内很好地证实了 DIM 改善 Treg 分化对 AhR 的依赖性和机制。
结论
DIM 通过增强 AhR 的激活以及随后的“糖酵解-乳酸-STAT3”和 TIP60 信号转导介导的 Treg 分化来增强 Treg 分化。
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