氧化应激下角质形成细胞中YTHDC1升高促进IL-1β表达  

YTHDC1 Mediates IL-1βExpresion in Keratinocyte under Oxidative Stress

作　　者：张喆 王琪[1] 陈佳希[1] 易秀丽 李舒丽[1] ZHANG Zhe;WANG Qi;CHEN Jia-xi;YIXiu-li;LI Shu-li(Department of Dennatology,Xijing Hospital,Fourth Military University,Xi'an,Shaanxi,710032,Chnia)

机构地区：[1]第四军医大学西京皮肤医院,陕西西安710032

出　　处：《现代生物医学进展》2022年第10期1811-1816,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81930087)。

摘　　要：目的:探讨氧化应激下角质形成细胞内m6A甲基化修饰酶YTHDC1异常对促炎因子的调控机制。方法:通过Western blot和q RT-PCR实验检测氧化应激下角质形成细胞中YTHDC1蛋白和m RNA表达水平。si RNA转染至角质形成细胞以干涉YTHDC1表达,随后继续给予300μM过氧化氢处理,通过Western blot和qRT-PCR实验检测角质形成细胞中促炎因子IL-1β蛋白和m RNA表达,进一步通过ELISA检测细胞上清中IL-1β分泌,通过CCK8法检测细胞存活水平。结果:1)过氧化氢刺激后人角质形成细胞系HaCaT细胞中YTHDC1表达水平较未处理组明显升高;2)干涉YTHDC1可以显著降低HaCaT细胞中IL-1β表达和上清中分泌;3)干涉YTHDC1后IL-1βm RNA稳定性下降,并且细胞存活率下降。结论:氧化应激下角质形成细胞中m6A甲基化修饰酶YTHDC1表达水平升高,通过提高m RNA稳定性促进IL-1β表达,可能是外界环境应激引起各种免疫性皮肤病的重要机制。Objective:To explore the regulatory mechanism of abnormal m6A methylation modifying enzyme YTHDC1 on pro-inflammatory factors in keratinocytes under oxidative stress.Methods:Western blot and qRT-PCR was used to detect the protein and m RNA expression of YTHDC1 in keratinocytes after the stimulation with H2O2.si RNA was transfected into keratinocytes to interfere with the expression of YTHDC1,and then treated with 300μM H2O2,the expression of pro-inflammatory factor IL-1βprotein and m RNA in keratinocytes was detected by Western blot and qRT-PCR.The secretion of IL-1βin the supernatant was further detected by ELISA,and the cell survival level was detected by CCK8.Results:1)H2O2increased the expression of YTHDC1 in HaCaT;2)The knockdown of YTHDC1 could significantly reduce the expression and secretion of IL-1βin HaCaT;3)IL-1βm RNA stability and cell viability decreased after YTHDC1 interference.Conclusions:m6A methylation modification enzyme YTHDC1 is involved in the stability of IL-1βm RNA in keratinocytes under oxidative stress,which may be an important mechanism of autoimmune skin diseases induced by external stress.

关 键 词：表观遗传修饰 m6A甲基化修饰 氧化应激 角质形成细胞 自身免疫性皮肤病 

分 类 号：R-33[医药卫生] R751