Wang Heng, Tian Qinqin, Zhang Ruijing, Du Qiujing, Hu Jie, Gao Tingting, Gao Siqi, Fan Keyi, Cheng Xing, Yan Sheng, Zheng Guoping, Dong Honglin
Department of Vascular Surgery, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, China.
Department of Nephrology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, China.
Lipids Health Dis. 2024 Mar 11;23(1):76. doi: 10.1186/s12944-024-02049-5.
Atherosclerosis (AS) is a persistent inflammatory condition triggered and exacerbated by several factors including lipid accumulation, endothelial dysfunction and macrophages infiltration. Nobiletin (NOB) has been reported to alleviate atherosclerosis; however, the underlying mechanism remains incompletely understood.
This study involved comprehensive bioinformatic analysis, including multidatabase target prediction; GO and KEGG enrichment analyses for function and pathway exploration; DeepSite and AutoDock for drug binding site prediction; and CIBERSORT for immune cell involvement. In addition, target intervention was verified via cell scratch assays, oil red O staining, ELISA, flow cytometry, qRT‒PCR and Western blotting. In addition, by establishing a mouse model of AS, it was demonstrated that NOB attenuated lipid accumulation and the extent of atherosclerotic lesions.
(1) Altogether, 141 potentially targetable genes were identified through which NOB could intervene in atherosclerosis. (2) Lipid and atherosclerosis, fluid shear stress and atherosclerosis may be the dominant pathways and potential mechanisms. (3) ALB, AKT1, CASP3 and 7 other genes were identified as the top 10 target genes. (4) Six genes, including PPARG, MMP9, SRC and 3 other genes, were related to the M0 fraction. (5) CD36 and PPARG were upregulated in atherosclerosis samples compared to the normal control. (6) By inhibiting lipid uptake in RAW264.7 cells, NOB prevents the formation of foam cell. (7) In RAW264.7 cells, the inhibitory effect of oxidized low-density lipoprotein on foam cells formation and lipid accumulation was closely associated with the PPARG signaling pathway. (8) In vivo validation showed that NOB significantly attenuated intra-arterial lipid accumulation and macrophage infiltration and reduced CD36 expression.
Nobiletin alleviates atherosclerosis by inhibiting lipid uptake via the PPARG/CD36 pathway.
动脉粥样硬化（AS）是一种持续性炎症状态，由脂质蓄积、内皮功能障碍和巨噬细胞浸润等多种因素引发并加重。据报道，川陈皮素（NOB）可缓解动脉粥样硬化；然而，其潜在机制仍未完全明确。
本研究涉及全面的生物信息学分析，包括多数据库靶点预测；用于功能和通路探索的基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析；用于药物结合位点预测的DeepSite和AutoDock；以及用于免疫细胞参与分析的CIBERSORT。此外，通过细胞划痕试验、油红O染色、酶联免疫吸附测定（ELISA）、流式细胞术、定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法对靶点干预进行了验证。另外，通过建立AS小鼠模型，证明NOB可减轻脂质蓄积和动脉粥样硬化病变程度。
（1）共鉴定出141个可能的可靶向基因，NOB可通过这些基因干预动脉粥样硬化。（2）脂质与动脉粥样硬化、流体切应力与动脉粥样硬化可能是主要通路及潜在机制。（3）白蛋白（ALB）、蛋白激酶B1（AKT1）、半胱天冬酶3（CASP3）等7个基因被确定为前10个靶基因。（4）过氧化物酶体增殖物激活受体γ（PPARG）、基质金属蛋白酶9（MMP9）、原癌基因酪氨酸蛋白激酶（SRC）等6个基因与M0细胞亚群相关。（5）与正常对照相比，动脉粥样硬化样本中CD36和PPARG表达上调。（6）通过抑制RAW264.7细胞中的脂质摄取，NOB可阻止泡沫细胞形成。（7）在RAW264.7细胞中，氧化型低密度脂蛋白对泡沫细胞形成和脂质蓄积的抑制作用与PPARG信号通路密切相关。（8）体内验证表明，NOB可显著减轻动脉内脂质蓄积和巨噬细胞浸润，并降低CD36表达。
川陈皮素通过PPARG/CD36途径抑制脂质摄取来减轻动脉粥样硬化。
