Melatonin ameliorates necrotizing enterocolitis by preventing Th17/Treg imbalance through activation of the AMPK/SIRT1 pathway
Melatonin ameliorates necrotizing enterocolitis by preventing Th17/Treg imbalance through activation of the AMPK/SIRT1 pathway
复制标题
褪黑素通过激活 AMPK/SIRT1 通路防止 Th17/Treg 失衡，从而改善坏死性小肠结肠炎
DOI：
10.7150/thno.45862
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Li, Sitao
中科院分区：
文献类型：
--
作者：
Ma, Fei;Hao, Hu;Li, Sitao
坏死性小肠结肠炎（NEC）是一种严重的胃肠道疾病，影响早产儿。越来越多的证据支持褪黑激素对NEC的治疗作用，尽管其潜在机制仍不清楚。研究方法：在10日龄C57 BL/6幼仔中通过缺氧和管饲含有肠道细菌的配方物诱导NEC，然后，小鼠接受褪黑激素、褪黑激素+重组IL-17、褪黑激素+抗CD 25单克隆抗体、褪黑激素+ Ex-527或褪黑激素+化合物C治疗。将对照小鼠与其母鼠一起哺乳，并将溶剂处理的NEC幼仔用作处理对照。从小鼠收集回肠组织，并通过组织病理学、免疫印迹和流式细胞术进行分析。将FITC标记的葡聚糖给予所有存活的幼仔，以通过荧光测定法评价肠道屏障功能。我们采用分子生物学和细胞培养的方法研究了脐血CD 4 + T细胞的相关机制。结果如下：我们证明了褪黑激素治疗以依赖于改善的肠道Th 17/Treg平衡的方式改善NEC小鼠模型中的疾病。我们还表明，褪黑激素阻断了致病性Th 17细胞的分化，并在体外增强了保护性Treg细胞的生成。我们进一步证明了褪黑激素通过激活肠道中的AMPK影响Th 17/Treg平衡，进而促进SIRT 1的激活和稳定。结论：这些结果表明，褪黑激素诱导的AMPK/SIRT 1信号转导激活调节Th 17和Treg细胞之间的平衡，并且通过AMPK/SIRT 1途径靶向Th 17/Treg平衡的治疗策略可能有利于NEC的治疗。
Necrotizing enterocolitis (NEC) is a severe gastrointestinal disease affecting premature infants. Mounting evidence supports the therapeutic effect of melatonin on NEC, although the underlying mechanisms remain unclear. Methods: NEC was induced in 10-day-old C57BL/6 pups via hypoxia and gavage feeding of formula containing enteric bacteria, and then, mice received melatonin, melatonin + recombinant IL-17, melatonin + anti-CD25 monoclonal antibody, melatonin + Ex-527, or melatonin + Compound C treatment. Control mice were left with their dams to breastfeed and vehicle-treated NEC pups were used as controls for treatment. Ileal tissues were collected from mice and analyzed by histopathology, immunoblotting, and flow cytometry. FITC-labeled dextran was administered to all surviving pups to evaluate gut barrier function by fluorometry. We used molecular biology and cell culture approaches to study the related mechanisms in CD4+ T cells from umbilical cord blood. Results: We demonstrated that melatonin treatment ameliorates disease in an NEC mouse model in a manner dependent on improved intestinal Th17/Treg balance. We also showed that melatonin blocks the differentiation of pathogenic Th17 cells and augments the generation of protective Treg cells in vitro. We further demonstrated that the Th17/Treg balance is influenced by melatonin through activation of AMPK in the intestine, in turn promoting SIRT1 activation and stabilization. Conclusions: These results demonstrate that melatonin-induced activation of AMPK/SIRT1 signaling regulates the balance between Th17 and Treg cells and that therapeutic strategies targeting the Th17/Treg balance via the AMPK/SIRT1 pathway might be beneficial for the treatment of NEC.