Typically inhibiting USP14 promotes autophagy in M1-like macrophages and alleviates CLP-induced sepsis.
Typically inhibiting USP14 promotes autophagy in M1-like macrophages and alleviates CLP-induced sepsis.
复制标题
通常抑制 USP14 会促进 M1 样巨噬细胞的自噬并减轻 CLP 诱导的脓毒症。
DOI：
10.1038/s41419-020-02898-9
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发表时间：
2020
影响因子：
9
通讯作者：
Wu Xudong
中科院分区：
文献类型：
--
作者：
Xu Fang;Ma Yuxiang;Huang Wei;Gao Jian;Guo Mengmeng;Li Jianxin;Kong Lingdong;Liang Guang;Du Ronghui;Xu Qiang;Wu Xudong
巨噬细胞是炎症性疾病的重要执行者，具有多种功能和多种表型。自噬已被证实与炎症的发生发展有着密切的联系，但其确切的调控机制尚不清楚，自噬调控因子在抗炎中的应用还有待进一步证实。本研究首次证实了新色胺S5（以下简称S5）可显著抑制M1样巨噬细胞的极化，并可降低促炎细胞因子的水平，下调NF-κB和STAT 1信号。然后，体内实验证明S5特别是基于M1样巨噬细胞的调节来改善盲肠结扎穿孔（CLP）诱导的脓毒症。机制研究表明，S5处理显着上调M1样巨噬细胞的细胞自噬。此外，通过多种方法发现，S5通过氢键直接与泛素特异性蛋白酶14（USP 14）在Ser 404、Phe 405和Cys 414处结合，从而抑制其去泛素化活性，并阻断USP 14-TRAF 6（TNF受体相关因子6）相互作用，随后促进Beclin 1的泛素化，中断Beclin 1-Bcl 2相互作用，自噬体在巨噬细胞中的积累，最终导致M1样巨噬细胞极化的阻断。动物实验也证实S5对CLP小鼠的保护作用依赖于巨噬细胞自噬的激活。S5作为一种新型的USP 14抑制剂，其抑制效率和安全性均优于已知的USP 14抑制剂IU 1。因此，本研究表明，通常抑制USP 14可促进M1样巨噬细胞中的自噬，并减轻CLP诱导的脓毒症。此外，我们提供了一种新的候选化合物S5，用于敏化自噬以干扰巨噬细胞炎症。
Macrophages, with diverse functions and variable phenotypes, are considered as an important executor of inflammatory diseases. And it has been proved that autophagy is deeply connected with the development of inflammation, while the exact regulatory mechanism still remains unclear, and the application of autophagy regulators in anti-inflammation needs to be further confirmed. Here, we firstly verified that neochromine S5 (hereinafter referred to as S5) significantly inhibited M1-like macrophage polarization with decrease of the proinflammatory cytokines and downregulation of NF-κB and STAT1 signals. Then, in vivo experiments demonstrated S5 improved cecal ligation and puncture (CLP)-induced sepsis specially based on the regulation of M1-like macrophages. Mechanistic studies indicated that S5 treatment dramatically upregulated cellular autophagy in M1-like macrophage. Furthermore, by multiple methods, S5 was revealed to directly bind with ubiquitin-specific proteases 14 (USP14) at Ser404, Phe405, and Cys414 by hydrogen bond to inhibit its deubiquitinating activity, and block USP14–TRAF6 (TNF receptor associated factor 6) interaction, subsequently promoting ubiquitination of Beclin1, interrupting Beclin1–Bcl2 interaction, and accumulating the autophagosome in macrophages, which finally resulted in the blockade of M1-like macrophage polarization. Animal experiments also confirmed the protection of S5 in CLP mice was dependent on activation of macrophage autophagy. What’s more, as a novel USP14 inhibitor, S5 exhibited higher efficiency and safety than IU1, the known USP14 inhibitor. Therefore, this study has demonstrated that typically inhibiting USP14 promotes autophagy in M1-like macrophages and alleviates CLP-induced sepsis. Moreover, we provide a new candidate compound, S5, for sensitizing autophagy to interfere with the macrophage inflammation.