Background Sepsis is a common severe complication in major burn victims and is characterized by a dysregulated systemic response to inflammation. YTH domain family 2 (YTHDF2), a well-studied N6-methyladenosine (m6A) reader that specifically recognizes and binds to m6A-modified transcripts to mediate their degradation, is connected to pathogenic and physiological processes in eukaryotes, but its effect on sepsis is still unknown. We aimed to discover the effects and mechanisms of YTHDF2 in sepsis. Methods Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot analyses were used to measure the expression of YTHDF2, the interleukin 6 receptor (IL-6R), high-mobility group box-1 (HMGB1), Janus kinase 2 (JAK2) and signal transducer and activator of transcription 1 (STAT1) under different in vitro conditions. Enzyme-linked immunosorbent assays were utilized to evaluate the expression of HMGB1, IL-6, IL-1β and tumor necrosis factor-α. To confirm that YTHDF2 specifically targets IL-6R mRNA, RNA immunoprecipitation and dual-luciferase reporter assays were performed. Finally, we utilized a mouse model of lipopolysaccharide (LPS)-induced sepsis to verify the effects of YTHDF2 in vivo. Results According to our findings, YTHDF2 was expressed at a low level in peripheral blood mononuclear cells from septic mice and patients as well as in LPS-induced RAW264.7 cells. Overexpression of YTHDF2 alleviated the inflammatory response by inhibiting HMGB1 release and JAK2/STAT1 signalling in LPS-stimulated cells. Mechanistically, YTHDF2 suppressed JAK2/STAT1 signalling by directly recognizing the m6A-modified site in IL-6R and decreasing the stability of IL-6R mRNA, thereby inhibiting HMGB1 release. In vivo experiments showed that YTHDF2 played a protective role in septic mice by suppressing the IL-6R/JAK2/STAT1/HMGB1 axis. Conclusions In summary, these findings demonstrate that YTHDF2 plays an essential role as an inhibitor of inflammation to reduce the release of HMGB1 by inhibiting the IL-6R/JAK2/STAT1 pathway, indicating that YTHDF2 is a novel target for therapeutic interventions in sepsis.

中文翻译：

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m6A阅读器YTHDF2通过抑制IL-6R/JAK2/STAT1通路介导的高迁移率group box-1释放来减轻炎症反应

背景脓毒症是严重烧伤患者常见的严重并发症，其特征是对炎症的全身反应失调。YTH 结构域家族 2 (YTHDF2) 是一种经过充分研究的 N6-甲基腺苷 (m6A) 阅读器，可特异性识别并结合 m6A 修饰的转录物以介导其降解，与真核生物的致病和生理过程有关，但其对脓毒症的影响是还是一个未知数。我们的目的是发现 YTHDF2 在脓毒症中的作用和机制。方法采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹分析检测YTHDF2、白细胞介素6受体（IL-6R）、高迁移率族盒-1（HMGB1）、Janus激酶2的表达(JAK2) 和信号转导子和转录激活子 1 (STAT1) 在不同的体外条件下。采用酶联免疫吸附测定法评估HMGB1、IL-6、IL-1β和肿瘤坏死因子-α的表达。为了确认 YTHDF2 特异性靶向 IL-6R mRNA，进行了 RNA 免疫沉淀和双荧光素酶报告基因测定。最后，我们利用脂多糖（LPS）诱导的脓毒症小鼠模型来验证YTHDF2在体内的作用。结果根据我们的研究结果，YTHDF2 在脓毒症小鼠和患者的外周血单核细胞以及 LPS 诱导的 RAW264.7 细胞中低水平表达。YTHDF2 的过表达通过抑制 LPS 刺激的细胞中 HMGB1 的释放和 JAK2/STAT1 信号传导来减轻炎症反应。从机制上讲，YTHDF2通过直接识别IL-6R中的m6A修饰位点并降低IL-6R mRNA的稳定性来抑制JAK2/STAT1信号传导，从而抑制HMGB1释放。体内实验表明，YTHDF2通过抑制IL-6R/JAK2/STAT1/HMGB1轴在脓毒症小鼠中发挥保护作用。结论 总之，这些研究结果表明，YTHDF2 作为炎症抑制剂发挥着重要作用，通过抑制 IL-6R/JAK2/STAT1 通路来减少 HMGB1 的释放，表明 YTHDF2 是脓毒症治疗干预的新靶点。