Rebalancing liver-infiltrating CCR3⁺ and CD206⁺ monocytes improves diet-induced NAFLD

Melatonin has been reported to improve nonalcoholic fatty liver disease (NAFLD), and exploring the underlying mechanisms will be beneficial for better treatment of NAFLD. Choline-deficient high-fat diet (CDHFD)- and methionine/choline-deficient diet (MCD)-fed mice with melatonin intervention exhibit significantly decreased liver steatosis, lobular inflammation, and focal liver necrosis. Single-cell RNA sequencing reveals that melatonin selectively inhibits pro-inflammatory CCR3⁺ monocyte-derived macrophages (MoMFs) and upregulates anti-inflammatory CD206⁺ MoMFs in NAFLD mice. Liver-infiltrating CCR3⁺CD14⁺ MoMFs are also significantly increased in patients with NAFLD. Mechanistically, melatonin receptor-independent BTG2-ATF4 signaling plays a role in the regulation of CCR3⁺ MoMF endoplasmic reticulum stress, survival, and inflammation. In contrast, melatonin upregulates CD206⁺ MoMF survival and polarization via MT1/2 receptors. Melatonin stimulation also regulates human CCR3⁺ MoMF and CD206⁺ MoMF survival and inflammation in vitro. Furthermore, CCR3 depletion antibody monotherapy inhibits liver inflammation and improves NAFLD in mice. Thus, therapies targeting CCR3⁺ MoMFs may have potential benefits in NAFLD treatment.