血管平滑肌细胞炎症的激活被认为是血管疾病的重要早期驱动因素。我们之前已经确定let-7 miRNA家族是动脉粥样硬化体外和体内模型中炎症的重要调节因子。在这里，我们研究了一种他汀类药物/let-7d-5p miRNA 联合治疗方法，以靶向人主动脉 SMC (HAoSMC) 激活和炎症。使用原代 HAoSMC 进行体外研究，以研究 let-7d-5p miRNA 过表达和抑制的影响。 HAoSMC 使用炎症细胞因子肿瘤坏死因子-α (TNF-α) 和阿托伐他汀或洛伐他汀联合治疗。 HAoSMC 的 Bulk RNA-seq 转录组学揭示了由 let-7d-5p miRNA 过表达和他汀类药物调节的下游调控网络。对 HAoSMC 进行蛋白质组分析细胞因子阵列、蛋白质印迹和定量 PCR 分析以验证关键发现。Let-7d-5p 过表达显着减弱 TNF-α 诱导的 IL-6、ICAM1、VCAM1、CCL2、CD68、MYOCD 基因表达的上调在 HAoSMC 中（p<0.05）。他汀类药物（阿托伐他汀、洛伐他汀）显着减弱 HAoSMC 中的炎症基因表达并上调 Let-7d 水平 (p<0.05)。 HAoSMC 中 Let-7d-5p 过表达/他汀类药物双重治疗的批量 RNA 测序分析表明，let-7d-5p 激活和他汀类药物在关键炎症通路（IL-6、IL-1β、TNF-α、IFN-γ）上汇聚）。 Let-7d-5p 过表达导致 ox-LDL 受体 OLR1 表达减少，这与 HAoSMC 中 ox-LDL 摄取降低相关。平滑肌细胞表型转换的计算机分析表明，HAoSMC 中 let-7d-5p 的过度表达可维持收缩表型。将 Let-7 网络与他汀类药物一起靶向可以调节 HAoSMC 激活并减弱关键炎症通路信号。版权所有 © 2024 作者。由 Elsevier B.V. 出版。保留所有权利。

Activation of vascular smooth muscle cell inflammation is recognised as an important early driver of vascular disease. We have previously identified the let-7 miRNA family as important regulators of inflammation in in vitro and in vivo models of atherosclerosis. Here we investigated a dual statin/let-7d-5p miRNA combination therapy approach to target human aortic SMC (HAoSMC) activation and inflammation.In vitro studies using primary HAoSMCs were performed to investigate the effects of let-7d-5p miRNA overexpression and inhibition. HAoSMCs were treated with combinations of the inflammatory cytokine tumor necrosis factor-α (TNF-α), and atorvastatin or lovastatin. HAoSMC Bulk RNA-seq transcriptomics of HAoSMCs revealed downstream regulatory networks modulated by let-7d-5p miRNA overexpression and statins. Proteome profiler cytokine array, Western blotting and quantitative PCR analyses were performed on HAoSMCs to validate key findings.Let-7d-5p overexpression significantly attenuated TNF-α-induced upregulation of IL-6, ICAM1, VCAM1, CCL2, CD68, MYOCD gene expression in HAoSMCs (p<0.05). Statins (atorvastatin, lovastatin) significantly attenuated inflammatory gene expression and upregulated Let-7d levels in HAoSMCs (p<0.05). Bulk RNA-seq analysis of a dual Let-7d-5p overexpression/statin therapy in HAoSMCs revealed that let-7d-5p activation and statins converge on key inflammatory pathways (IL-6, IL-1β, TNF-α, IFN-γ). Let-7d-5p overexpression led to reduced expression of the ox-LDL receptor OLR1, and this was associated with lower ox-LDL uptake in HAoSMCs. In silico analysis of smooth muscle cell phenotypic switching shows that overexpression of let-7d-5p in HAoSMCs maintains a contractile phenotype.Targeting the Let-7 network alongside statins can modulate HAoSMC activation and attenuate key inflammatory pathway signals.Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.