MicroRNA-124-3p inhibits collagen synthesis in atherosclerotic plaques by targeting prolyl 4-hydroxylase subunit alpha-1 (P4HA1) in vascular smooth muscle cells
MicroRNA-124-3p inhibits collagen synthesis in atherosclerotic plaques by targeting prolyl 4-hydroxylase subunit alpha-1 (P4HA1) in vascular smooth muscle cells
复制标题
MicroRNA-124-3p 通过靶向血管平滑肌细胞中的脯氨酰 4-羟化酶亚基 α-1 (P4HA1) 来抑制动脉粥样硬化斑块中的胶原蛋白合成。
DOI：
10.1016/j.atherosclerosis.2018.08.034
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发表时间：
2018-10-01
期刊：
影响因子：
5.3
通讯作者：
Zhang, Mei
中科院分区：
文献类型：
--
作者：
Chen, Wei'jia;Yu, Fangpu;Zhang, Mei
背景和目的：血管平滑肌细胞（VSMCs）中的胶原合成在动脉粥样硬化中非常重要，因为它影响斑块的稳定性。在这项研究中，我们的目的是评估是否miR-124- 3 p参与VSMCs的胶原合成过程中，它可能发挥的作用在atherosclerosis development.Methods：我们调制miR-124- 3 p的表达在主动脉根部斑块高脂饮食喂养ApoE（-/-）小鼠慢病毒注射。为了确定斑块大小和斑块稳定性相关细胞或分子的含量，进行染色，包括苏木精和伊红、油红O、天狼星红和免疫组织化学染色。采用荧光原位杂交（FISH）技术对动脉粥样硬化斑块中miR-124- 3 p进行定位。采用Western blotting和RT-qPCR方法检测P4 HA 1、I型胶原和III型胶原蛋白及mRNA的表达水平。结果：斑块中胶原和VSMC含量与miR-124- 3 p水平呈负相关。通过荧光原位杂交（FISH），我们发现miR-124- 3 p主要在VSMCs中表达。我们还发现miR-124- 3 p降低了动脉粥样硬化斑块和动脉粥样硬化斑块中I型和III型胶原蛋白的蛋白水平。我们在体外调节miR-124- 3 p的水平，发现它可以抑制HASMCs的胶原表达。这可能与P4 HA 1的下调有关。结论：miR-124 - 3 p通过直接靶向P4 HA 1抑制VSMC胶原合成，从而降低动脉粥样硬化斑块的稳定性。
Background and aims: Collagen synthesis in vascular smooth muscle cells (VSMCs) is very important in atherosclerosis, as it affects plaque stability. In this study, we aim to assess whether miR-124-3p is involved in the collagen synthesis process in VSMCs and the role it might play in atherosclerotic development.Methods: We modulated the miR-124-3p expression in the aortic root plaques of high-fat-diet fed ApoE(-/-) mice by lentivirus injection. To determine plaque size and the content of plaque-stability-related cells or molecules, stainings, including hematoxylin and eosin, Oil red O, Sirius Red and immunohistochemical staining, were performed. Fluorescence in situ hybridization (FISH) was used to locate miR-124-3p in atherosclerotic plaques. Western blotting and RT-qPCR were carried out to determine the level of P4HA1 as well as type I and type III collagen protein and mRNA expression.Results: Results showed that collagen and VSMC content of plaques was inversely correlated with miR-124-3p levels. By FISH, we identified that miR-124-3p was primarily expressed by VSMCs. We also found that protein levels of type I and type III collagen in aortas and atherosclerotic plaques were decreased by miR-124-3p. We modulated miR-124-3p level in vitro and found it could inhibit collagen expression in HASMCs. This might be caused by the downregulation of P4HA1. P4HA1 was predicted as miR-124-3p's direct target, which was verified with a dual luciferase reporter assay and RIP test.Conclusions: The results presented here provide evidence that miR-124-3p inhibits VSMC collagen synthesis by directly targeting P4HA1, which might decrease atherosclerotic plaque stability.