Oncostatin M sensitizes keratinocytes to UVB-induced inflammation via GSDME-mediated pyroptosis
Oncostatin M sensitizes keratinocytes to UVB-induced inflammation via GSDME-mediated pyroptosis
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DOI：
10.1016/j.jdermsci.2021.09.004
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发表时间：
2021-11-27
影响因子：
4.6
通讯作者：
Yang, Bin
中科院分区：
文献类型：
--
作者：
Liu, Jun;Zhong, Yadan;Yang, Bin
背景：肿瘤抑制素M （Oncostatin M， OSM）是一种白细胞介素-6 （IL-6）家族促炎细胞因子，在炎症性皮肤病中起重要作用，但其作用机制尚不清楚。目的：探讨OSM诱导正常人表皮角质形成细胞（NHEKs）和永生化人角质形成细胞（HaCaT）热沉的机制。方法：用OSM处理NHEKs和HaCaT细胞。利用CRISPR/ Cas9系统敲除OSM受体（OSMR），用小干扰RNA敲除GSDME和OSMR -/-和GSDME -/-小鼠的原代角质形成细胞，研究OSMR和GSDME的作用。OSM处理后，用UVB照射NHEKs和HaCaT细胞。采用实时荧光定量聚合酶链反应（qRT-PCR）和RNA测序法分析mRNA， Western Blotting、Elisa和免疫荧光法检测蛋白水平。乳酸脱氢酶（LDH）释放检测细胞死亡情况。结果：我们发现OSM诱导NHEKs和HaCaT细胞热沉，但敲除OSMR可消除热沉。RNA测序结果显示，OSM治疗后参与NLRP3炎性小体激活的几个关键基因上调，其中NLRP3、GSDME和IL-1 β通过qRT-PCR和Western Blotting证实。GSDME基因敲低可减轻osm诱导的焦腐病。OSM预处理增强了uvb诱导的NHEKs和HaCaT细胞的焦亡和炎症，而这种启动功能在Osmr-/-和Gsdme-/-小鼠的角化细胞中丧失。在人体表皮的三维培养中也获得了类似的结果。结论：OSM作为一种启动细胞因子，可增强uvb诱导的角质形成细胞炎症，为炎性皮肤病的发病机制提供线索。(C) 2021日本皮肤病研究学会。Elsevier B.V.版权所有。
Background: Oncostatin M (OSM), an interleukin-6 (IL-6) family proinflammatory cytokine, plays a critical role in inflammatory skin diseases, but its mechanism of action is not well understood.Objective: To demonstrate the mechanism of OSM induced pyropotosis in normal human epidermal keratinocytes (NHEKs) and immortalized human keratinocytes (HaCaT cells).Methods: NHEKs and HaCaT cells were treated with OSM. Knockout of OSM receptor (OSMR) with CRISPR/ Cas9 system, knockdown of GSDME with small interfering RNA and primary keratinocytes from Osmr-/- and Gsdme-/- mice were used to study the effect of OSMR and GSDME. After treatment of OSM, NHEKs and HaCaT cells were irradiated with UVB. The mRNA was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and RNA sequencing, protein level was detected by Western Blotting, Elisa and immunofluorescence. Cell death was examined by lactate dehydrogenase (LDH) releasing.Results: Here we found that OSM induced pyropotosis in NHEKs and HaCaT cells, but knockout of OSMR abolished pyropotosis. RNA sequencing revealed an upregulation of several key genes involved in NLRP3 inflammasome activation following OSM treatment, among which NLRP3, GSDME, and IL-1 beta were confirmed by qRT-PCR and Western Blotting. Knockdown of GSDME alleviated OSM-induced pyropotosis. Pretreatment of OSM boosted UVB-induced pyroptosis and inflammation in NHEKs and HaCaT cells, and this priming function was lost in keratinocytes of Osmr-/- and Gsdme-/- mice. Similar results were obtained in a 3-dimensional culture of human epidermis.Conclusion: OSM functions as a priming cytokine to enhance UVB-induced inflammation in keratinocytes, providing insight into the pathogenesis of inflammatory skin diseases. (C) 2021 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.