摘要
为探讨miR-128-3p对THP-1源性泡沫细胞形成的影响,采用佛波醇12-肉豆蔻酸酯13-醋酸盐(phorbol 12-myristate 13-acetate,PMA)将THP-1细胞诱导分化成巨噬细胞并分为对照组、氧化低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)组、miR-NC组、miR-128-3p mimic组、miR-128-3p mimic+核苷酸结合寡聚化结构域蛋白1(nucleotide-binding oligomerization domain-containing protein 1,NOD1)激动剂(iE-DAP)组。给予相应的转染处理后,用ox-LDL(100μg/mL)诱导泡沫细胞形成。检测细胞中miR-128-3p、NOD1 mRNA表达和脂质积累;检测细胞中胆固醇含量和细胞培养上清液中IL-6、TNF-α水平;检测细胞中NOD1、受体相互作用蛋白2(receptor-interacting protein 2,RIP2)、磷酸化RIP2(phosphorylated RIP2,p-RIP2)、NF-κB p65(p65)、磷酸化p65(phosphorylated p65,p-p65)蛋白表达;验证miR-128-3p与NOD1的靶向关系。结果显示,与对照组比较,ox-LDL组miR-128-3p水平降低(P<0.05),脂质含量,胆固醇含量,NOD1表达,p-RIP2/RIP2、p-p65/p65比值以及IL-6、TNF-α水平升高(P<0.05);与ox-LDL组比较,miR-128-3p mimic组miR-128-3p表达升高(P<0.05),NOD1表达,RIP2、p65的磷酸化,脂质、胆固醇积累以及TNF-α、IL-6水平降低(P<0.05);与miR-128-3p mimic组比较,miR-128-3p mimic+iE-DAP组NOD1表达,RIP2、p65的磷酸化,脂质、胆固醇积累以及TNF-α、IL-6水平升高(P<0.05);与转染miR-NC比较,转染miR-128-3p mimic后,含有NOD1-WT的荧光素酶报告基因的活性降低(P<0.05)。该研究提示,过表达miR-128-3p可能通过抑制NOD1/RIP2信号通路抑制THP-1源性泡沫细胞形成。
The purpose of this study is to investigate the influence of miR-128-3p on the formation of THP-1-derived foam cells.To this end,THP-1 cells were induced to differentiate into macrophages by phorbol 12-myristate 13-acetate(PMA)and divided into control group,oxidized low-density lipoprotein(ox-LDL)group,miR-NC group,miR-128-3p mimic group,and miR-128-3p mimic+nucleotide-binding oligomerization domain-containing protein 1(NOD1)agonist(iE-DAP)group.Foam cell formation was induced by ox-LDL(100μg/mL)after the corresponding transfection treatment.The expressions of miR-128-3p,NOD1 mRNA,and lipid accumulation in cells were examined and the content of cholesterol in cells and the levels of TNF-αand IL-6 in cell culture supernatants were detected.Protein expressions of NOD1,receptor-interacting protein 2(RIP2),phosphorylated RIP2(p-RIP2),NF-κB p65(p65),and phosphorylated p65(p-p65)in cells were measured.The targeting between miR-128-3p and NOD1 was validated.The results showed that compared to those of the control group,miR-128-3p level in the ox-LDL group was decreased(P<0.05),while the lipid content,cholesterol content,NOD1 mRNA and protein levels,p-RIP2/RIP2 and p-p65/p65 ratios,and the TNF-αand IL-6 levels in the cells were all increased(P<0.05).Compared to those of the ox-LDL group,the expression of miR-128-3p in the miR-128-3p mimic group was increased(P<0.05),while the expression of NOD1,the phosphorylation of RIP2 and p65,the accumulation of lipid and cholesterol,and the levels of TNF-αand IL-6 were all decreased(P<0.05).Compared to those of the miR-128-3p mimic group,the expression of NOD1,the phosphorylation of RIP2 and p65,the accumulation of lipid and cholesterol,and the levels of TNF-αand IL-6 were all increased in the miR-128-3p mimic+iE-DAP group(P<0.05).Compared to cells transfected with miR-NC,cells transfected with miR-128-3p mimic showed decreased activity of luciferase reporter gene containing NOD1-WT(P<0.05).This study suggests that overexpression of miR-128-3p may inhibit the formation of THP-
作者
聂俊丽
谈宝珍
侯亮
韩静
NIE Jun-li;TAN Bao-zhen;HOU Liang;HAN Jing(Internal Medicine-Cardiovascular Department,General Hospital of the Yangtze River Shipping,Wuhan 430000,China)
出处
《现代免疫学》
CAS
2024年第3期227-233,共7页
Current Immunology
基金
武汉市卫生健康委科研计划资助项目(WX20Q10)。
