摘要
目的探讨吲哚胺2,3-双加氧酶(IDO)基因缺失对PO诱导的高尿酸血症动脉粥样硬化小鼠模型疾病进展的影响及其分子机制。方法在体实验使用遗传背景为C57BL/6J的ApoE^(-/-)小鼠(ApoE^(-/-)组)或ApoE^(-/-)IDO^(-/-)(ApoE^(-/-)IDO^(-/-)组)小鼠,给予其高脂富含胆固醇饮食联合腹腔注射PO诱导构建高尿酸血症-动脉粥样硬化小鼠模型,对照组为野生型(WT组)小鼠给予正常啮齿类饲料;ELISA法测定小鼠血浆总胆固醇、甘油三酯、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、血清尿酸、犬尿氨酸(Kyn)、色氨酸(Trp)并Kyn/Trp比率;油红O染色分析小鼠动脉粥样硬化斑块面积。体外实验分离野生型或IDO^(-/-)小鼠原代巨噬细胞,给予LPS处理,通过Western blot法测定巨噬细胞自噬相关蛋白Parkin、p62、LC3-II/I和NLRP3炎症小体相关蛋白NLRP3、Caspase-1和Pro-IL-1β的表达水平。结果与ApoE^(-/-)组相比,ApoE^(-/-)IDO^(-/-)组小鼠血浆总胆固醇、甘油三酯和LDL-C明显升高;血清尿酸、HDL-C和Kyn/Trp(%)明显降低(P<0.05);油红O染色显示,与ApoE^(-/-)组小鼠相比,ApoE^(-/-)IDO^(-/-)组小鼠的血管动脉粥样硬化斑块面积明显增加(P<0.05)。体外研究显示,与WT+LPS组原代巨噬细胞相比,IDO^(-/-)+LPS组细胞自噬相关蛋白Parkin和LC3-II/LC3-I以及NLRP3炎症小体相关蛋白NLRP3、Caspase-1和Pro-IL-1β的表达水平明显升高,p62的表达水平明显降低(P<0.05)。结论IDO表达缺失将增强巨噬细胞内自噬和NLRP3炎症小体的水平加重高尿酸血症-动脉粥样硬化小鼠的疾病进展。
Objective To investigate the effect of indoleamine 2,3-dioxygenase(IDO)gene deletion on disease progression of PO-induced hyperuricemia-atherosclerosis mouse model and its molecular mechanisms.Methods In vivo ex-periments used ApoE^(-/-)mice(ApoE^(-/-)group)or ApoE^(-/-)IDO^(-/-)(ApoE^(-/-)IDO^(-/-)group)mice with a genetic background of C57BL/6J,given a high-fat,cholesterol-rich diet combined with intraperitoneal injection of PO,induced to construct a hy-peruricemia-atherosclerosis mouse model.Control group was wild-type(WT group)mice given normal rodent chow diet.ELI-SA methods was used to determine the total cholesterol,triglyceride,high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),serum uric acid,kynurenine(Kyn),tryptophan(Trp)and Kyn/Trp ratio.Oil red O staining analysis of atherosclerotic plaque area in mice.Wild-type or IDO^(-/-)mouse primary macrophages were isolated in vi-tro and treated with LPS,and the autophagy-related proteins Parkin,p62,LC3-II/I and NLRP3 inflammasomes related pro-reins NLRP3,Caspase-1 and Pro-IL-1βwere detected by western blot.Results Compared with the ApoE^(-/-)group,the plasma total cholesterol,triglyceride and LDL-C of the ApoE^(-/-)IDO^(-/-)group mice were significantly increased;serum uric ac-id,HDL-C and Kyn/Trp(%)were significantly increased.decreased(P<0.05);Oil red O staining showed that compared with the ApoE^(-/-)group mice,the vascular atherosclerotic plaque area of the ApoE^(-/-)IDO^(-/-)group mice was significantly in-creased(P<0.05).In vitro studies showed that compared with the primary macrophages in the WT+LPS group,the autopha-gy-related proteins Parkin and LC3-II/LC3-I and the NLRP3 inflammasomes-related protein NLRP3,Caspase-1 and Pro-IL-1βin the IDO^(-/-)+LPS group were significantly increased,and the expression level of p62 was significantly decreased(P<0.05).Conclusions Loss of IDO expression enhances autophagy and NLRP3 inflammasomes’level in macrophages and ag-gravates disease progression in hyperuricemic-atherosclerotic mice.
作者
刘超
岳阳阳
LIU Chao;YUE Yang-yang(Department of vascular surgery,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an,Shaanxi Province 710061,China)
出处
《解剖学研究》
CAS
2022年第6期551-557,565,共8页
Anatomy Research
