UBC9介导SUMO化修饰在同型半胱氨酸诱导巨噬细胞焦亡中的作用研究  

Role of UBC9-mediated SUMO modification in homocysteine-induced pyroptosis of macrophages

作　　者：马凌桔 迟宏扬 吴昕雪 马福军 田彦成 赵彩齐 何天宇 彭红建 姜怡邓 杨力[1] 黄晖[1] 马胜超 MA Lingju;CHI Hongyang;WU Xinxue;MA Fujun;TIAN Yancheng;ZHAO Caiqi;HE Tianyu;PENG Hongjian;JIANG Yideng;YANG Li;HUANG Hui;MA Shengchao(Department of Geriatrics and Special Needs Medicine,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission,Yinchuan 750004;School of Laboratory Medicine,Ningxia Medical University,Yinchuan 750004;Basic School of Medicine,Ningxia Medical University,Yinchuan 750004)

机构地区：[1]宁夏医科大学总医院老年与特需医学科,银川750004 [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,银川750004 [3]宁夏医科大学检验学院,银川750004 [4]宁夏医科大学基础医学院,银川750004

出　　处：《中国比较医学杂志》2024年第6期11-17,共7页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金(81900273,82270492);宁夏青年科技拔尖人才培养项目;宁夏自然科学基金优秀青年项目(2023AAC05035)

摘　　要：目的探讨泛素结合酶9(ubiquitin conjugating enzyme 9,UBC9)介导SUMO化修饰在同型半胱氨酸诱导巨噬细胞焦亡中的作用。方法首先采用细胞计数试剂(cell counting kit-8,CCK-8)法和Western blot检测不同浓度(0μmol/L、50μmol/L、100μmol/L、150μmol/L和200μmol/L)同型半胱氨酸对小鼠巨噬细胞(RAW264.7)的活力及焦亡的影响;采用Western blot检测不同组细胞中UBC9、SUMO化修饰蛋白SUMO-1、炎症因子IL-1β的表达水平;qRT-PCR检测干扰前后UBC9的mRNA表达,同时检测干扰UBC9后,UBC9、焦亡相关蛋白、IL-1β及SUMO-1相关表达。结果当采用100μmol/L的Hcy刺激后,巨噬细胞的活力影响最小且焦亡蛋白NLRP3、Caspase-1表达最明显(P<0.001);与Control组相比,Hcy组IL-1β的表达水平升高(P<0.01),SUMO-1表达升高(P<0.01);与Control组相比,Hcy组中UBC9蛋白水平及mRNA水平表达均增高(P<0.05);转染si-UBC9后,与si-NC+Hcy组相比,si-UBC9+Hcy组NLRP3、Caspase-1、IL-1β、UBC9、SUMO-1表达降低(P<0.01)。结论同型半胱氨酸诱导巨噬细胞焦亡的发生,其机制与上调泛素结合酶9发生SUMO化修饰有关。ObjectiveTo study the role of ubiquitin-conjugating enzyme 9(UBC9)in thepyroptosisof homocysteine-induced macrophages mediated by small ubiquitin-like modifier(SUMO)modification.Methods First,the effects of homocysteine at different concentrations(0μmol/L,50μmol/L,100μumol/L,150μmol/L and 200μmol/L)on the viability and pyrodeath of mouse macrophages(RAW264.7)were detected by CCK-8 and Western blot.Western blot was used to detect the expression levels of UBC9,SUMO-1,and the inflammatory cytokine IL-1βin different groups of cells.qRT-PCR was used to detect the mRNA expression of UBC9 before and after RNA interference and the expression of UBC9,pyrogen-related protein,and SUMO-1 after RNA interference.Results After stimulation with 100μmol/L homocysteine,the effect of macrophage activity was minimal,and NLRP3 and Caspase-1 were the proteins with the most obvious increase in expression(P<0.05).Compared with the Control group,the Hcy group's expression of IL-1βand SUMO-1 was increased(P<0.01).Compared with the Control group,the Hcy group's UBC9 protein and mRNA levels were increased(P<0.05).The expression of NLRP3,Caspase-1,IL-1β,UBC9,and SUMO-1 was decreased in the si-UBC9+Hcy group compared with the si-NC+Hey group(P<0.01).Conclusions Homocysteine induces pyroptosis in macrophages,and its mechanism of action is related to the up-regulation of UBC9 to induce SUMO modification.

关 键 词：UBC9 SUMO化 巨噬细胞 同型半胱氨酸 细胞焦亡 

分 类 号：R-33[医药卫生]