MDA-5 DNA甲基化在同型半胱氨酸致巨噬细胞炎症中的作用研究  

Role of MDA-5 DNA methylation in Hcy-induced macrophage inflammation

作　　者：陈勇 杨亚丽 张玲 田荣 袁茵 杨晓玲 CHEN Yong;YANG Ya-li;ZHANG Ling;TIAN Rong;YUAN Yin;YANG Xiao-ling(Department of Cardiology,Yinchuan Second People’s Hospital,Yinchuan,Ningxia 750011,China;不详)

机构地区：[1]银川市第二人民医院心内科,宁夏银川750011 [2]国家卫生健康委员会代谢性心血管疾病研究重点实验室,宁夏银川750004 [3]宁夏医科大学基础医学院,宁夏银川750004

出　　处：《现代预防医学》2021年第16期3024-3028,共5页Modern Preventive Medicine

基　　金：国家自然科学基金(81760095)。

摘　　要：目的探讨MDA-5在同型半胱氨酸(Hcy)致巨噬细胞炎症反应中的作用及机制。方法巨噬细胞用Hcy干预24 h,ELISA检测TNF-α和IL-6含量;荧光定量PCR和western-blot检测MDA-5、DNA甲基转移酶(DNMTs)的m RNA和蛋白表达;巢式降落式甲基化特异性PCR检测MDA-5启动子区甲基化水平;将MDA-5重组腺病毒(AdMDA-5)和干扰片段(si-MDA-5)转染巨噬细胞检测细胞内TNF-α和IL-6含量。结果 Hcy干预细胞后,细胞内TNF-α和IL-6含量分别增加了37.5%(t=2.243 0,P=0.035 4)和25%(t=2.464,P=0.031 4),MDA-5的mRNA和蛋白表达也分别增加了52.5%(t=3.522,P=0.012 5)和41.3%(t=3.183,P=0.019 0),而MDA-5启动子区甲基化水平降低了30.1%(t=3.377,P=0.027 9),DNMT3a表达降低了32.7%(t=3.258,P=0.031 1);过表达MDA-5后,细胞内TNF-α和IL-6含量比Hcy组分别增加了60.1%(t=2.625,P=0.039 3)和64.6%(t=8.867,P=0.000 1),而干扰MDA-5的表达,TNF-α和IL-6含量分别比Hcy组降低了13.6%(t=4.188,P=0.005 8)和23.6%(t=3.958,P=0.007 2)。结论MDA-5 DNA低甲基化介导了Hcy诱导的巨噬细胞炎症反应。Objective To investigate the role and mechanism of MDA-5 in the inflammatory response of macrophages induced by homocysteine.Methods Macrophages were treated with Hcy for 24 hours,TNF-and IL-6 were detected by ELISA,mRNA and protein expression of MDA-5 and DNA methyltransferases were detected by fluorescence quantitative PCR and Western Blot.The promoter methylation level of MDA-5 was detected by nested drop methylation-specific PCR.The contents of TNF-αand IL-6 were detected by transfection of MDA-5 recombinant adenovirus(Ad-MDA-5)and interference fragment(si-MDA-5)into macrophages.Results After Hcy treatment,the contents of TNF-and IL-6 were significantly increased by37.5%(t=2.2430,P=0.0354)and 25%(t=3.706,P=0.0035),and the mRNA and protein expression of MDA-5 were also significantly increased by 52.5%(t=3.522,P=0.0125)and 41.3%(t=3.183,P=0.0190).The level of methylation in promoter region of MDA-5 and the protein expression of DNMT3 a were decreased by 30.1%(t=3.377,P=0.0279)and 32.7%(t=3.258,P=0.0311).Compared with Hcy group,the contents of TNF-αand IL-6 were increased by 60.1%(t=2.625,P=0.0393)and 64.6%(t=8.867,P=0.0001).When MDA-5 was interfered,the contents of TNF-and IL-6 decreased by 13.6%(t=4.188,P=0.0058)and 23.6%(t=3.958,P=0.0072).Conclusion Low methylation of MDA-5 DNA mediates the inflammatory response of macrophages induced by Hcy.

关 键 词：MDA-5 DNA甲基化 同型半胱氨酸 炎症 

分 类 号：R363.21[医药卫生—病理学]