Theranostics 2020; 10(17):7730-7746. doi:10.7150/thno.45862

Research Paper

Melatonin ameliorates necrotizing enterocolitis by preventing Th17/Treg imbalance through activation of the AMPK/SIRT1 pathway

Fei Ma1,2*, Hu Hao1*, Xiaoyan Gao3, Yao Cai1, Jialiang Zhou4, Puping Liang5, Junjian Lv6, Qiuming He6, Congcong Shi1, Dandan Hu6, Bowei Chen7, Lixin Zhu2✉, Xin Xiao1✉, Sitao Li1✉

1. Department of Pediatrics, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510655, China.
2. Guangdong Institute of Gastroenterology, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510655, China.
3. Department of Neonatology, the Foshan Women and Children hospital, Foshan, 528000, China.
4. Department of Neonatal Surgery, Guangdong Women and Children Hospital, Guangzhou, 511400, China.
5. MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University, Guangzhou, 510275, China.
6. Congenital Diaphragmatic Hernia Study and Collaborative Group of Fetal Care Center, Municipal Women and Children's Medical Center, Guangzhou, 510623, China.
7. Zhongshan School of medicine, Sun Yat-sen University, Guangzhou, 510080, China.
*These authors contributed equally to this work.

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Citation:
Ma F, Hao H, Gao X, Cai Y, Zhou J, Liang P, Lv J, He Q, Shi C, Hu D, Chen B, Zhu L, Xiao X, Li S. Melatonin ameliorates necrotizing enterocolitis by preventing Th17/Treg imbalance through activation of the AMPK/SIRT1 pathway. Theranostics 2020; 10(17):7730-7746. doi:10.7150/thno.45862. Available from http://www.thno.org/v10p7730.htm

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Abstract

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.

Keywords: melatonin, necrotizing enterocolitis, Th17/Treg imbalance, AMPK/SIRT1 pathway, intestine