作者：王婷婷，张鹏珂，史敏，于倩怡，乔娇，薛玉刚

单位：

710032陕西省西安市，空军军医大学唐都医院心内科

单位（英文）：

Department of Cardiology, Tangdu Hospital, Air Force Medical University, Xi'an 710032, China

关键词：

动脉粥样硬化；巨噬细胞；脂类代谢；异鼠李素；自噬

关键词（英文）：

Atherosclerosis; Macrophages; Lipid metabolism; Isorhamnetin; Autophagy

中图分类号：

R 543.5

DOI：

10.12114/j.issn.1008-5971.2023.00.237

基金项目：

国家自然科学基金资助项目（82200404）

摘要：

目的　探讨异鼠李素预防巨噬细胞脂质代谢紊乱的可能机制。方法　本实验时间为2022年1—6月。取对数生长期的RAW264.7细胞，将其分为对照组（不干预）、动脉粥样硬化组〔采用50 μg/ml的氧化低密度脂蛋白（ox-LDL）干预24 h〕、动脉粥样硬化+异鼠李素组（采用20 μmol/L的异鼠李素孵育8 h，之后采用50 μg/ml的ox-LDL干预24 h）。采用油红O染色检测各组巨噬细胞中脂质含量，Western blot法检测各组巨噬细胞中自噬标志物〔微管相关蛋白1轻链3（LC3Ⅱ）、P62〕表达水平，透射电镜观察各组巨噬细胞中自噬小体数目。结果　油红O染色结果显示，对照组巨噬细胞中未发现脂质沉积；动脉粥样硬化组巨噬细胞中存在明显的脂质沉积现象；与动脉粥样硬化组比较，动脉粥样硬化+异鼠李素组巨噬细胞中脂质含量明显减少。动脉粥样硬化组、动脉粥样硬化+异鼠李素组巨噬细胞中LC3Ⅱ表达水平低于对照组，P62表达水平高于对照组（P＜0.05）；动脉粥样硬化+异鼠李素组巨噬细胞中LC3Ⅱ表达水平高于动脉粥样硬化组，P62表达水平低于动脉粥样硬化组（P＜0.05）。透射电镜观察结果显示，对照组巨噬细胞中可观察到自噬小体；与对照组相比，动脉粥样硬化组和动脉粥样硬化+异鼠李素组巨噬细胞中自噬小体数目明显减少；与动脉粥样硬化组相比，动脉粥样硬化+异鼠李素组巨噬细胞中自噬小体数目增多。结论　异鼠李素可能通过上调自噬水平来预防巨噬细胞脂质代谢紊乱，进而预防泡沫细胞形成。

英文摘要：

Objective To investigate the specific mechanism of isorhamnetin in preventing lipid metabolism disorderin macrophages. Methods This study was conducted from January to June 2022. RAW264.7 cells in logarithmic growthphase were divided into control group (no intervention) , atherosclerosis group [treated with 50 μg/ml oxidized low-densitylipoprotein (ox-LDL) for 24 h] , atherosclerosis+isorhamnetin group (incubated with 20 μmol/L isorhamnetin for 8 h, and thentreated with 50 μg/ml ox-LDL for 24 h) . The lipid content of macrophages in each group was detected by oil red O staining.The expression level of autophagy markers [microtubule-associated protein 1 light chain 3 (LC3) Ⅱ and P62] in macrophagesin each group was detected by Western blot. The number of autophagosomes in macrophages in each group was observed bytransmission electron microscopy. Results Oil red O staining showed that no lipid deposition was found in macrophages ofcontrol group; there was obvious lipid deposition in macrophages of atherosclerosis group; the lipid content of macrophages in theatherosclerosis+isorhamnin group was significantly reduced compared with that in the atherosclerosis group. The expression levelof LC3Ⅱ in macrophages of atherosclerosis group and atherosclerosis+isorhamnetin group was lower than that of control group,and the expression level of P62 was higher than that of control group (P < 0.05) . The expression level of LC3Ⅱ in macrophagesof atherosclerosis+isorhamnin group was higher than that of atherosclerosis group, and the expression level of P62 was lowerthan that of atherosclerosis group (P < 0.05) . The results of transmission electron microscopy showed that autophagosomescould be observed in macrophages of the control group; compared with the control group, the number of autophagosomes in theatherosclerosis group and the atherosclerosis+isorhamnetin group was significantly reduced; compared with atherosclerosis group,the number of autophagosomes in macrophages of atherosclerosis+isorhamnin group was increased. Conclusion Isorhamnetin can prevent the lipid metabolism disorder in macrophages by up-regulating autophagy level, and then prevent the formation offoam cells.

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