The First Affiliated Hospital of Jinan University, Guangzhou, China.
The Second Affiliated Hospital of Bengbu Medical College, Bengbu, China.
Funct Integr Genomics. 2023 Mar 7;23(2):77. doi: 10.1007/s10142-023-01001-7.
Atherosclerosis (AS) is the main cause of cardiovascular diseases. However, the role of AQP9 in AS is not well understood. In the present study, we predicted that miR-330-3p might regulate AQP9 in AS through bioinformatics analysis, and we established AS model using ApoE mouse (C57BL/6) with high-fat diet (HFD). Hematoxylin and eosin (H&E) and Oil red O staining were used to determine atherosclerotic lesions. CCK8 and Ethyny1-2-deoxyuridine (EdU) assays were used to investigate human umbilical vein endothelial cells (HUVECs) proliferation after treatment with 100 μg/mL ox-LDL. Wound scratch healing and transwell assays were used to measure the cell invasion and migration ability. Flow cytometry assay was used to determine apoptosis and cell cycle. A dual-luciferase reporter assay was performed to investigate the binding of miR-330-3p and AQP9. We identified that the expression of miR-330-3p in AS mice model decreased while the expression level of AQP9 increased. miR-330-3p overexpression or down-regulation of AQP9 could reduce cell apoptosis, promote cell proliferation, and migration after ox-LDL treatment. Dual-luciferase reporter assay result presented that AQP9 was directly inhibited by miR-330-3p. These results suggest that miR-330-3p inhibits AS by regulating AQP9. miR-330-3p/AQP9 axis may be a new therapeutic target for AS.
动脉粥样硬化（AS）是心血管疾病的主要病因。然而，AQP9 在 AS 中的作用尚不清楚。在本研究中，我们通过生物信息学分析预测 miR-330-3p 可能通过调控 AQP9 参与 AS 的发生，通过高脂饮食（HFD）喂养 ApoE 基因敲除小鼠（C57BL/6）建立 AS 模型。采用苏木精和伊红（H&E）及油红 O 染色法检测动脉粥样硬化病变。用 CCK8 法和 Ethyny1-2-deoxyuridine（EdU）法检测经 100μg/ml ox-LDL 处理后人脐静脉内皮细胞（HUVECs）的增殖情况。采用划痕愈合实验和 Transwell 小室实验检测细胞侵袭和迁移能力。流式细胞术检测细胞凋亡和细胞周期。双荧光素酶报告实验检测 miR-330-3p 与 AQP9 的结合情况。结果表明，AS 小鼠模型中 miR-330-3p 的表达降低，AQP9 的表达升高。过表达 miR-330-3p 或下调 AQP9 可减少 ox-LDL 处理后细胞凋亡，促进细胞增殖和迁移。双荧光素酶报告实验结果表明 AQP9 受 miR-330-3p 直接抑制。这些结果提示 miR-330-3p 通过调控 AQP9 抑制 AS。miR-330-3p/AQP9 轴可能成为 AS 的一个新的治疗靶点。
