Department of Cardiology, The People's Hospital of China Three Gorges University/The First People's Hospital of Yichang, Yichang, China.
Department of Cardiology, Shanghai Songjiang District Center Hospital, Shanghai, China.
J Cell Physiol. 2019 Apr;234(4):5319-5326. doi: 10.1002/jcp.27340. Epub 2018 Sep 27.
Atherosclerosis has been recognized as a chronic inflammation process induced by lipid of the vessel wall. Oxidized low-density lipoprotein (ox-LDL) can drive atherosclerosis progression involving macrophages. Recently, long noncoding RNAs (lncRNAs) have been reported to play critical roles in atherosclerosis development. In our current study, we focused on the biological roles of lncRNA NEAT1 in atherosclerosis progress. Here, we found that ox-LDL was able to trigger human macrophages THP-1 cells, a human monocytic cell line, apoptosis in a dose-dependent and time-dependent course. In addition, we observed that NEAT1 was significantly increased in THP-1 cells incubated with ox-LDL and meanwhile miR-342-3p was greatly decreased. Then, NEAT1 was silenced by transfection of small interfering RNA (siRNA) of NEAT1 into THP-1 cells. As exhibited, CD36, oil-red staining levels, total cholesterol (TC), total cholesterol (TG) levels and THP-1 cell apoptosis were obviously repressed by knockdown of NEAT1. Furthermore, inhibition of NEAT1 contributed to the repression of inflammation in vitro. Interleukin 6 (IL-6), IL-1β, cyclooxygenase-2 (COX-2) and tumour necrosis factor-alpha (TNF-α) protein levels were remarkably depressed by NEAT1 siRNA in THP-1 cells. By using bioinformatics analysis, miR-342-3p was predicted as a downstream target of NEAT1 and the correlation between them was confirmed in our study. Moreover, overexpression of miR-342-3p could also greatly suppress inflammation response and lipid uptake in THP-1 cells. Knockdown of NEAT1 and miR-342-3p mimics inhibited lipid uptake in THP-1 cells. In conclusion, we implied that blockade of NEAT1 repressed inflammation response through modulating miR-342-3p in human macrophages THP-1 cells and NEAT1 may offer a promising strategy to treat atherosclerotic cardiovascular diseases.
动脉粥样硬化已被认为是一种由血管壁脂质引起的慢性炎症过程。氧化型低密度脂蛋白（ox-LDL）可驱动涉及巨噬细胞的动脉粥样硬化进展。最近，长链非编码 RNA（lncRNA）已被报道在动脉粥样硬化发展中发挥关键作用。在我们目前的研究中，我们专注于 lncRNA NEAT1 在动脉粥样硬化进展中的生物学作用。在这里，我们发现 ox-LDL 能够以剂量和时间依赖的方式触发人单核细胞 THP-1 细胞，一种人单核细胞系，细胞凋亡。此外，我们观察到 ox-LDL 孵育的 THP-1 细胞中 NEAT1 显著增加，同时 miR-342-3p 大大减少。然后，通过转染 NEAT1 的小干扰 RNA（siRNA）将 NEAT1 沉默转染到 THP-1 细胞中。结果表明，通过敲低 NEAT1，CD36、油红染色水平、总胆固醇（TC）、总胆固醇（TG）水平和 THP-1 细胞凋亡明显受到抑制。此外，抑制 NEAT1 有助于体外抑制炎症。白细胞介素 6（IL-6）、白细胞介素 1β（IL-1β）、环氧化酶-2（COX-2）和肿瘤坏死因子-α（TNF-α）蛋白水平在 THP-1 细胞中通过 NEAT1 siRNA 显著降低。通过生物信息学分析，miR-342-3p 被预测为 NEAT1 的下游靶标，并且在本研究中证实了它们之间的相关性。此外，过表达 miR-342-3p 也可以大大抑制 THP-1 细胞中的炎症反应和脂质摄取。THP-1 细胞中敲低 NEAT1 和 miR-342-3p 模拟物抑制脂质摄取。总之，我们暗示阻断 NEAT1 通过调节人巨噬细胞 THP-1 细胞中的 miR-342-3p 来抑制炎症反应，并且 NEAT1 可能为治疗动脉粥样硬化性心血管疾病提供有前途的策略。
