Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS) & Comparative Medicine Centre, Peking Union Medical College(PUMC), Beijing Collaborative Innovation Center for Cardiovascular Disorders, Beijing Engineering Research Center for Experimental Animal Models of Human Critical Diseases, Beijing 100021, China.
School of Basic Medical Sciences, Ningxia Medical University, 750004 Yinchuan, China.
Life Sci. 2021 Jan 15;265:118816. doi: 10.1016/j.lfs.2020.118816. Epub 2020 Dec 2.
Endothelial to mesenchymal transition (EndMT) is closely related to atherosclerosis. Herein, we aim to determine whether miR-122 is involved in EndMT and the underlying mechanism in atherosclerosis.
qRT-PCR was performed to detect miR-122 expression in ApoE mice and cellular EndMT model induced by HO. MiR-122 expression in vivo was modulated by lenti-virus injection and by genetic manipulation. Hematoxylin and eosin (HE) and Oil-red O staining were used to observe the plaque size and lipid accumulation in the aortic roots. F4/80 staining, elastin staining, and masson staining were used to observe the components of atherosclerotic lesions. MiR-122 expression in endothelial cells was modulated by transfection of miR-122 mimic and inhibitor. Western blotting and co-localization of endothelial markers (VE-cadherin, CD31) and mesenchymal markers (Vimentin, α-SMA) were carried out to determine EndMT.
MiR-122 was upregulated in the aortic intima and serum of ApoE mice induced by HFD and in cellular EndMT model. Inhibition of miR-122 repressed the atherosclerotic plaque progression and vulnerable plaque formation in ApoE mice. In vitro, endothelial cells acquired a spindle-shaped morphology accompanying decrease of the endothelial markers (VE-cadherin, CD31) and increase of the mesenchymal markers (Vimentin, α-SMA) in the presence of HO, which was inhibited by miR-122 inhibitor. Furthermore, NPAS3 functions as a target of miR-122, and NPAS3 silencing abolished the anti-EndMT effect of miR-122 inhibitor.
Inhibition of miR-122 prevents atherosclerosis and regulates NPAS3-mediated EndMT, suggesting that miR-122 may be a novel target in the treatment of EndMT-associated diseases including atherosclerosis.
内皮到间充质转化（EndMT）与动脉粥样硬化密切相关。在此，我们旨在确定 miR-122 是否参与动脉粥样硬化中的 EndMT 及其潜在机制。
通过 qRT-PCR 检测 ApoE 小鼠和 HO 诱导的细胞内 EndMT 模型中 miR-122 的表达。通过慢病毒注射和基因操作调节体内 miR-122 的表达。苏木精和伊红（HE）及油红 O 染色观察主动脉根部斑块大小和脂质积聚。F4/80 染色、弹力蛋白染色和 Masson 染色观察动脉粥样硬化病变的成分。通过转染 miR-122 模拟物和抑制剂调节内皮细胞中 miR-122 的表达。通过 Western blot 和内皮标志物（VE-cadherin、CD31）和间充质标志物（Vimentin、α-SMA）的共定位来确定 EndMT。
在 HFD 诱导的 ApoE 小鼠主动脉内膜和血清中以及细胞内 EndMT 模型中，miR-122 上调。抑制 miR-122 可抑制 ApoE 小鼠动脉粥样硬化斑块的进展和易损斑块的形成。在体外，HO 存在下，内皮细胞获得梭形形态，伴随内皮标志物（VE-cadherin、CD31）减少和间充质标志物（Vimentin、α-SMA）增加，miR-122 抑制剂可抑制这种变化。此外，NPAS3 是 miR-122 的靶标，NPAS3 沉默消除了 miR-122 抑制剂的抗 EndMT 作用。
抑制 miR-122 可预防动脉粥样硬化，并调节 NPAS3 介导的 EndMT，提示 miR-122 可能成为包括动脉粥样硬化在内的与 EndMT 相关疾病的治疗新靶点。
