Quercetin Suppresses the Progression of Atherosclerosis by Regulating MST1-Mediated Autophagy in ox-LDL-Induced RAW264.7 Macrophage Foam Cells
Quercetin Suppresses the Progression of Atherosclerosis by Regulating MST1-Mediated Autophagy in ox-LDL-Induced RAW264.7 Macrophage Foam Cells
复制标题
槲皮素通过调节 ox-LDL 诱导的 RAW264.7 巨噬细胞泡沫细胞中 MST1 介导的自噬来抑制动脉粥样硬化的进展
DOI：
10.3390/ijms20236093
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发表时间：
2019-12-01
影响因子：
5.6
通讯作者：
Shen, Dingzhu
中科院分区：
文献类型：
--
作者：
Cao, Hui;Jia, Qingling;Shen, Dingzhu
目的：通过上调RAW264.7巨噬细胞MST1介导的自噬作用，探讨其抑制动脉粥样硬化的作用机制。方法：用氧化型低密度脂蛋白(ox-LDL)培养RAW264.7巨噬细胞，建立体外泡沫细胞模型。这些细胞被单独或与自噬抑制剂3-甲基腺嘌呤和自噬激动剂雷帕霉素联合处理。CCK-8试剂盒检测细胞活性。油红O染色检测脂质聚集，SA-β-GAL(衰老相关β-半乳糖苷酶)染色检测衰老，ROS试剂盒检测活性氧。用透射电子显微镜观察自噬小体和线粒体，免疫荧光和免疫印迹法检测MST1、LC3-II/I、Beclin1、Bcl2、P21和P16的表达。结果：氧化低密度脂蛋白可诱导RAW264.7巨噬细胞源性泡沫细胞形成，降低细胞存活率，加重细胞脂质堆积，诱导细胞出现衰老表型。伴随而来的是自噬小体形成减少，P53、P21和P16表达增加，Lc3-II/I和Beclin1表达降低。经槲皮素干预后，细胞存活率提高，脂质堆积减少，衰老表型减少。此外，Lc3-II/I和Beclin1的表达增加，这与栎素促进自噬的能力是一致的。氧化低密度脂蛋白还可增加MST1的表达，这种增加可被槲皮素阻断，这提供了一种潜在的机制，即槲皮素可能通过其保护泡沫细胞免受与年龄相关的有害影响。结论：槲皮素能抑制氧化低密度脂蛋白诱导的泡沫细胞形成，延缓衰老。其机制可能与调节MST1介导的RAW264.7细胞自噬有关。
Objective: To investigate the process by which quercetin suppresses atherosclerosis by upregulating MST1-mediated autophagy in RAW264.7 macrophages. Methods: An in vitro foam cell model was established by culturing RAW264.7 macrophages with oxidized low-density lipoprotein (ox-LDL). The cells were treated with quercetin alone or in combination with the autophagy inhibitor, 3-methyladenine, and autophagy agonist, rapamycin. Cell viability was detected with a CCK-8 kit. Lipid accumulation was detected by oil red O staining, senescence was detected by SA-β-gal (senescence-associated β-galactosidase) staining, reactive oxygen species were detected by ROS assay kit. Autophagosomes and mitochondria were detected by transmission electron microscope (TEM), and expression of MST1, LC3-II/I, Beclin1, Bcl-2, P21, and P16 were detected by immunofluorescence and Western blot. Results: Ox-LDL induced RAW264.7 macrophage-derived foam cell formation, reduced survival, aggravated cell lipid accumulation, and induced a senescence phenotype. This was accompanied by decreased formation of autophagosome; increased expression of P53, P21, and P16; and decreased expression of LC3-II/I and Beclin1. After intervention with quercetin, the cell survival rate was increased, and lipid accumulation and senescence phenotype were reduced. Furthermore, the expression of LC3-II/I and Beclin1 were increased, which was consistent with the ability of quercetin to promote autophagy. Ox-LDL also increased the expression of MST1, and this increase was blocked by quercetin, which provided a potential mechanism by which quercetin may protect foam cells against age-related detrimental effects. Conclusion: Quercetin can inhibit the formation of foam cells induced by ox-LDL and delay senescence. The mechanism may be related to the regulation of MST1-mediated autophagy of RAW264.7 cells.