血管芯片中早期动脉粥样硬化的 RNA 测序分析及其在比较可燃卷烟和加热烟草制品中的应用

IF 2.9 Q2 TOXICOLOGY
Kazuhiro Ohashi , Ayaka Hayashida , Atsuko Nozawa, Shigeaki Ito
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引用次数: 0

摘要

我们之前的研究表明,当血管内皮细胞(VEC)通过M0巨噬细胞暴露于香烟烟雾(CS)提取物时,复制早期动脉粥样硬化的结果很有希望。动脉粥样硬化是一种涉及血管内皮细胞和免疫细胞交流的复杂疾病,我们使用了器官芯片系统作为动物实验的替代方法来模拟动脉粥样硬化。通过将巨噬细胞纳入血管芯片系统,我们旨在模拟 CS 等可吸入物质对血管内皮细胞的间接影响。在本研究中,我们通过对直接或通过巨噬细胞间接暴露于 CS 的血管内皮细胞进行转录组分析,进一步检验了我们的体外系统是否适合模拟早期动脉粥样硬化。我们还加入了 M1 巨噬细胞,以复制预先存在的炎症状态。与间接暴露方法相比,我们在直接暴露方法中发现了更多的差异表达基因(DEGs)。然而,通路分析表明,CS 直接暴露于 VECs 主要引起细胞死亡相关通路的改变,而 "动脉粥样硬化信号 "通路预计会受到负调控。通过 M0 巨噬细胞的间接暴露也同样表明,已确定的 DEGs 与细胞死亡有关,而 "动脉粥样硬化信号 "通路预计会被激活。与此相反,通过 M1 巨噬细胞将 CS 间接暴露于血管内皮细胞时,未观察到与细胞死亡相关的通路改变,但通路扰动与促炎症阳性对照相似。此外,"动脉粥样硬化信号 "通路预计会在通过 M1 巨噬细胞间接暴露于 CS 的 VEC 中被激活。这些结果表明,M0 或 M1 巨噬细胞对血管内皮细胞致动脉粥样硬化转录组变化有贡献,尽管它们对细胞死亡相关通路的影响不同。我们还使用间接暴露方法比较了 CS 和加热烟草制品(HTP)气溶胶的影响。值得注意的是,在暴露于 HTP 气溶胶的血管内皮细胞中,与动脉粥样硬化相关的基因表达变化不如 CS 那么明显。我们的研究强调了血管芯片系统通过巨噬细胞间接接触 CS 提取物复制动脉粥样硬化的实用性,并表明 HTP 有降低风险的潜力。这项研究有助于推动毒理学和疾病模型研究中动物试验的替代方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

RNA sequencing analysis of early-stage atherosclerosis in vascular-on-a-chip and its application for comparing combustible cigarettes with heated tobacco products

RNA sequencing analysis of early-stage atherosclerosis in vascular-on-a-chip and its application for comparing combustible cigarettes with heated tobacco products

Our previous study showed promising results in replicating early-stage atherosclerosis when vascular endothelial cells (VECs) were exposed to cigarette smoke (CS) extract via M0 macrophages. We used an organ-on-a-chip system as an alternative to animal testing to model atherosclerosis, which is a complex disease involving endothelial and immune cell communications. By incorporating macrophages into the vascular-on-a-chip system, we aimed to mimic the indirect effects of inhalable substances, such as CS, on VECs. In the current study, we further examined the suitability of our in vitro system for mimicking early-stage atherosclerosis by transcriptomic analyses of VECs exposed to CS directly or indirectly via macrophages. We also incorporated M1 macrophages to replicate a preexisting inflammatory state. We found a greater number of differentially expressed genes (DEGs) in direct exposure methods than indirect exposure methods. However, a pathway analysis showed that the direct exposure of CS to VECs primarily caused cell death-related pathway alterations, and the “Atherosclerosis Signaling” pathway was predicted to be negatively regulated. Indirect exposure via M0 macrophages similarly showed that the identified DEGs were related to cell death, while the “Atherosclerosis Signaling” pathway was predicted to be activated. In contrast, cell death-related pathway alterations were not observed by indirect exposure of CS to VECs via M1 macrophages, but the pathway perturbations were similar to a pro-inflammatory positive control. In addition, the “Atherosclerosis Signaling” pathway was predicted to be activated in VECs that were indirectly exposed to CS via M1 macrophages. These results suggest that M0 or M1 macrophages contribute to atherogenic transcriptomic changes in VECs, although they affect cell death-related pathways differently. We also used indirect exposure methods to compare the effects of CS and heated tobacco product (HTP) aerosol. Notably, gene expression changes related to atherosclerosis were less pronounced in HTP aerosol-exposed VECs than CS. Our study highlights the utility of the vascular-on-a-chip system with indirect exposure of CS extract via macrophages for replicating atherogenesis and suggests a reduced risk potential of the HTP. This research contributes to advancing alternatives to animal testing for toxicological and disease modeling studies.

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来源期刊
Current Research in Toxicology
Current Research in Toxicology Environmental Science-Health, Toxicology and Mutagenesis
CiteScore
4.70
自引率
3.00%
发文量
33
审稿时长
82 days
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