Department of Dermatology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Engineering Research Center of Traditional Chinese Medicine Intelligent Rehabilitation, Ministry of Education, Shanghai, China.
J Cell Mol Med. 2024 Feb;28(3):e18085. doi: 10.1111/jcmm.18085. Epub 2023 Dec 25.
Interleukin-6 (IL-6) is a cytokine generated by healthy constituents of the skin, but is also up-regulated by a wide range of skin lesions and inflammatory conditions to trigger cytopathy of skin cells. TRIM27 was identified to contribute to the functional effects of IL-6 on skin cells. However, the underlying mechanism was not clear. Lentivirus infection was used for gene overexpression or silencing. RT-PCR and Western blot were used to respectively assess mRNA and protein levels. Cell viability was assessed by CCK-8 assay. Extracellular flux analysis was used to assess the levels of oxygen consumption rate and extracellular acidification rate. Mouse back skin was treated with imiquimod to produce psoriasis-like inflammation in vivo. Histological assessment and immunohistochemistry staining were respectively applied to analyse lesioned mouse and human skin samples. IL-6-induced increased viability, glycolysis and inflammation in keratinocytes was inhibited both by a chemical methylation inhibitor and by METTL14 knockdown. Further investigation found that METTL14 induces m6A methylation of TRIM27, which is recognized by a m6A reader, IGF2BP2. Elevation of TRIM27 level and activation of IL-6/STAT3 signalling pathway were found in an in vivo psoriasis-like inflammation model, whereas inhibition m6A methylation strongly alleviated the inflammation. Finally, METTL14, TRIM27, STAT3, p-STAT3 and IL-6 expressions were all found to be increased in clinical skin samples of psoriatic patients. Our results unravelled METTL14/TRIM27/IGF2BP2 signalling axis in keratinocyte cytopathy, which plays a critical role in facilitating the activation of IL-6/STAT3 signalling pathway. Our findings should provide inspirations for the design of new therapeutics for skin inflammatory diseases including psoriasis.
白细胞介素 6 (IL-6) 是皮肤健康成分产生的细胞因子,但也被广泛的皮肤损伤和炎症状态上调,以触发皮肤细胞的细胞病变。TRIM27 被确定为 IL-6 对皮肤细胞的功能效应作出贡献。然而,其潜在的机制尚不清楚。采用慢病毒感染进行基因过表达或沉默。分别采用 RT-PCR 和 Western blot 检测 mRNA 和蛋白水平。通过 CCK-8 测定评估细胞活力。采用外通量分析评估耗氧率和细胞外酸化率的水平。用咪喹莫特处理小鼠背部皮肤,在体内产生银屑病样炎症。分别应用组织学评估和免疫组化染色分析病变小鼠和人皮肤样本。IL-6 诱导的角质形成细胞活力增加、糖酵解和炎症均被化学甲基化抑制剂和 METTL14 敲低所抑制。进一步的研究发现,METTL14 诱导 TRIM27 的 m6A 甲基化,被 m6A 阅读器 IGF2BP2 识别。在体内银屑病样炎症模型中发现 TRIM27 水平升高和 IL-6/STAT3 信号通路激活,而抑制 m6A 甲基化则强烈缓解炎症。最后,在银屑病患者的临床皮肤样本中发现 METTL14、TRIM27、STAT3、p-STAT3 和 IL-6 的表达均增加。我们的结果揭示了角质形成细胞病变中 METTL14/TRIM27/IGF2BP2 信号轴,在促进 IL-6/STAT3 信号通路的激活中起着关键作用。我们的研究结果应为设计包括银屑病在内的皮肤炎症性疾病的新疗法提供启示。