Musashi-2 通过与 Fbxo6 结合诱导 Rnaset2 泛素化，从而在动脉粥样硬化中的血管平滑肌细胞表型转换中诱导趋化因子信号通路。
Musashi-2 binds with Fbxo6 to induce Rnaset2 ubiquitination and chemokine signaling pathway during vascular smooth muscle cell phenotypic switch in atherosclerosis.
机构信息
Department of General Surgery, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan 250014, Shandong, China; Department of General Surgery, The First Hospital Affiliated with Shandong First Medical University, Jinan 250014, Shandong, China.
Department of General Surgery, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan 250014, Shandong, China; Department of General Surgery, The First Hospital Affiliated with Shandong First Medical University, Jinan 250014, Shandong, China.
出版信息
Cell Signal. 2023 Nov;111:110869. doi: 10.1016/j.cellsig.2023.110869. Epub 2023 Aug 24.
OBJECTIVE
The objective of this study is to determine how Musashi-2 (MSI2) affects vascular smooth muscle cell (VSMC) phenotypic switch and contributes to atherosclerosis (AS).
METHODS
Primary mouse VSMCs were transfected with MSI2 specific siRNA and treated with platelet-derived growth factor-BB (PDGF-BB). The proliferation, cell-cycle, and migration of VSMCs were determined by CCK-8, flow cytometry, wound healing, and transwell assays. Western blot and qRT-PCR were conducted to analyze the protein and mRNA expression. Moreover, the correlation between MSI2, Fbxo6, Rnaset2, and chemokine signaling was predicted and verified using RNAct database, KEGG, wiki, RNA-binding protein immunoprecipitation and co-immunoprecipitation. Moreover, H&E and Oil Red O staining were employed for assessing necrotic core and lipid accumulation in AS mouse aorta tissues. The numbers of B lymphocytes and monocytes, and the levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDLC), and low-density lipoprotein cholesterol (LDL-C) in AS mice blood were investigated using flow cytometry and corresponding commercial kits, respectively.
RESULTS
MSI2 was up-regulated in the PDGF-BB-treated VSMCs. Knockdown of MSI2 inhibited VSMC proliferation, cell-cycle, and migration. Moreover, MSI2 regulated VSMC phenotypic switch through binding with Fbxo6 to induce Rnaset2 ubiquitination. MSI2 knockdown inhibited chemokine signaling via regulating Fbxo6/Rnaset2 axis. In AS mice, knockdown of MSI2 inhibited the formation of necrotic core and atherosclerotic plaque, and inhibited chemokine signaling via regulating Fbxo6/Rnaset2 axis.
CONCLUSION
Our findings demonstrated that MSI2 could bind with Fbxo6 to induce Rnaset2 ubiquitination and the activation of chemokine signaling pathway during VSMC phenotypic switch in AS.
目的
本研究旨在探讨 Musashi-2（MSI2）如何影响血管平滑肌细胞（VSMC）表型转换并促进动脉粥样硬化（AS）。
方法
用 MSI2 特异性 siRNA 转染原代小鼠 VSMC，并用血小板衍生生长因子-BB（PDGF-BB）处理。通过 CCK-8、流式细胞术、划痕愈合和 Transwell 测定法测定 VSMC 的增殖、细胞周期和迁移。Western blot 和 qRT-PCR 用于分析蛋白质和 mRNA 表达。此外，使用 RNAct 数据库、KEGG、wiki、RNA 结合蛋白免疫沉淀和共免疫沉淀预测和验证 MSI2、Fbxo6、Rnaset2 和趋化因子信号之间的相关性。此外，H&E 和油红 O 染色用于评估 AS 小鼠主动脉组织中的坏死核心和脂质积累。使用流式细胞术和相应的商业试剂盒分别检测 AS 小鼠血液中的 B 淋巴细胞和单核细胞数量以及甘油三酯（TG）、总胆固醇（TC）、高密度脂蛋白胆固醇（HDLC）和低密度脂蛋白胆固醇（LDL-C）水平。
结果
MSI2 在 PDGF-BB 处理的 VSMC 中上调。MSI2 敲低抑制 VSMC 增殖、细胞周期和迁移。此外，MSI2 通过与 Fbxo6 结合诱导 Rnaset2 泛素化来调节 VSMC 表型转换。MSI2 敲低通过调节 Fbxo6/Rnaset2 轴抑制趋化因子信号。在 AS 小鼠中，MSI2 敲低抑制坏死核心和动脉粥样硬化斑块的形成，并通过调节 Fbxo6/Rnaset2 轴抑制趋化因子信号。
结论
我们的研究结果表明，MSI2 可以与 Fbxo6 结合，在 AS 中 VSMC 表型转换过程中诱导 Rnaset2 泛素化和趋化因子信号通路的激活。
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