Wenjie Luo, Yubin Chen, Cheng Fang, Hui Shi, Fanyan Luo
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引用次数: 0
Abstract
Objectives: Atherosclerosis is characterized by persistent inflammatory condition, leading to various cardiovascular complications. Foam cell formation, resulting from macrophage uptake of oxidized low-density lipoprotein (ox-LDL), contributes significantly to atherosclerosis progression. This study was designed to investigate the involvement of bispecific phosphatase-6 (DUSP6) and its potential regulatory mechanisms in foam cell formation and atherosclerosis.
Methods: We employed THP-1 cells to induce foam cell formation. The lipid droplet accumulation, cholesterol content, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 levels were evaluated using Oil Red O staining, cholesterol assay, ELISA, and qRT-PCR techniques. We investigated DUSP6 ubiquitination via immunoprecipitation and western blot (WB) analysis. A bioinformatics approach identified FBXL14 as a potential E3 ligase involved in DUSP6 ubiquitination, further confirmed by siRNA and overexpression experiments. The impact of FBXL14 on the NRF2 signaling pathway was assessed using WB analysis.
Results: DUSP6 interference suppressed foam cell formation and inflammatory factor secretion. Upon ox-LDL treatment, DUSP6 underwent deubiquitylation, with FBXL14 emerging as the candidate E3 ligase. FBXL14 overexpression induced DUSP6 ubiquitination, leading to the NRF2 signaling pathway activation. It counteracted with DUSP6 overexpression on foam cell formation and inflammation. In ApoE-/- mice, sh-DUSP6 adenovirus injection mitigated atherosclerotic lesion progression and improved the lipid profile, with increased the proteins expression of NQO1, HO-1, and NRF2 in aortic tissue.
Conclusion: DUSP6 and FBXL14 play vital roles in modulating foam cell formation and inflammatory responses in atherosclerosis. Targeting these molecules could offer therapeutic potential in attenuating atherosclerosis-related complications.
目的：动脉粥样硬化的特点是持续的炎症状态，导致各种心血管并发症。泡沫细胞的形成，由巨噬细胞摄取氧化低密度脂蛋白（ox-LDL），显著促进动脉粥样硬化的进展。本研究旨在探讨双特异性磷酸酶-6 （DUSP6）在泡沫细胞形成和动脉粥样硬化中的作用及其潜在的调节机制。方法：采用THP-1细胞诱导泡沫细胞形成。采用油红O染色、胆固醇测定、ELISA和qRT-PCR技术评估脂滴积累、胆固醇含量、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β和IL-6水平。我们通过免疫沉淀和western blot （WB）分析DUSP6的泛素化。生物信息学方法鉴定FBXL14是参与DUSP6泛素化的潜在E3连接酶，并通过siRNA和过表达实验进一步证实。利用WB分析评估FBXL14对NRF2信号通路的影响。结果：DUSP6干扰抑制泡沫细胞形成和炎症因子分泌。在ox-LDL处理后，DUSP6发生去泛素化，FBXL14成为候选E3连接酶。FBXL14过表达诱导DUSP6泛素化，导致NRF2信号通路激活。抑制了DUSP6过表达对泡沫细胞形成和炎症的影响。在ApoE-/-小鼠中，注射sh-DUSP6腺病毒可减轻动脉粥样硬化病变进展，改善脂质谱，并增加主动脉组织中NQO1、HO-1和NRF2的蛋白表达。结论：DUSP6和FBXL14在动脉粥样硬化中调控泡沫细胞形成和炎症反应中发挥重要作用。靶向这些分子可能在减轻动脉粥样硬化相关并发症方面提供治疗潜力。临床试验号：不适用。
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