Table_3_Long Noncoding RNA-H19 Contributes to Atherosclerosis and Induces Ischemic Stroke via the Upregulation of Acid Phosphatase 5.xlsx

Objective: Atherosclerosis is closely associated with ischemic stroke, and long noncoding RNA-H19 (lncRNA-H19) might be a potential target for treating atherosclerosis. The present study aimed to investigate the function of lncRNA-H19 in atherosclerosis and to explore a novel therapeutic strategy for ischemic stroke.
Methods: Differentially expressed genes (DEGs) in atherosclerosis were screened by searching public database. In combination with the lncRNA-H19-knockout database, potential lncRNA-H19-mediated gene was retrieved and their relationship was identified. In order to assess the detailed regulatory mechanism of lncRNA-H19, we used a lentivirus packaging system to upregulate Acp5 (Acid phosphatase 5) expression in vascular smooth muscle cells (VSMC) and human umbilical vein endothelial cells (HUVECs). The expression of ACP5 was determined by Western Blot, and evaluations of cell proliferation and apoptosis were detected. An ischemic stroke mouse model was established. Atherosclerosis was measured by using plaque area size. The effects H19 on the expression of ACP5 were explored by the overexpression or silence of H19.
Results: H19 and ACP5 were associated with Acute Stroke Treatment (TOAST) subtypes of atherosclerotic patients. The target prediction program and dual-luciferase reporter confirmed ACP5 as a direct target of H19. Lentivirus-mediated H19-forced expression upregulated ACP5 protein levels, promoted cell proliferation and suppressed the apoptosis. The plaque area size was larger in ischemic models than controls. The overexpression or silence of H19 increased or reduced the plaque size. The overexpression or silence of H19 resulted in the expression or inhibition of ACP5.
Conclusion: IncRNA-H19 promoting ACP5 protein expression contributed to atherosclerosis and increased the risk of ischemic stroke.

研究目的：动脉粥样硬化（Atherosclerosis）与缺血性脑卒中（ischemic stroke）密切相关，长链非编码RNA-H19（long noncoding RNA-H19, lncRNA-H19）或可成为治疗动脉粥样硬化的潜在靶点。本研究旨在探讨长链非编码RNA-H19在动脉粥样硬化中的作用，并探索缺血性脑卒中的新型治疗策略。
研究方法：通过检索公共数据库筛选动脉粥样硬化相关差异表达基因（Differentially expressed genes, DEGs）。结合长链非编码RNA-H19敲除数据库，检索得到长链非编码RNA-H19介导的潜在靶基因，并明确其相互作用关系。为解析长链非编码RNA-H19的具体调控机制，本研究采用慢病毒包装系统（lentivirus packaging system），在血管平滑肌细胞（vascular smooth muscle cells, VSMC）与人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVECs）中上调酸性磷酸酶5（Acid phosphatase 5, Acp5）的表达。通过Western Blot检测ACP5的表达水平，并开展细胞增殖与凋亡相关评估。构建缺血性脑卒中小鼠模型，通过测定斑块面积评估动脉粥样硬化程度。通过过表达或沉默H19，探究H19对ACP5表达的调控作用。
研究结果：H19与ACP5与动脉粥样硬化患者的急性卒中治疗（Acute Stroke Treatment, TOAST）分型相关。靶基因预测程序（target prediction program）与双荧光素酶报告基因实验（dual-luciferase reporter）证实ACP5为H19的直接靶标。慢病毒介导的H19过表达可上调ACP5蛋白水平，促进细胞增殖并抑制细胞凋亡。缺血性脑卒中模型小鼠的斑块面积较对照组更大。过表达或沉默H19可分别增大或缩小斑块面积。H19的过表达或沉默分别可上调或抑制ACP5的表达。
研究结论：长链非编码RNA-H19通过促进ACP5蛋白表达，参与动脉粥样硬化进程并增加缺血性脑卒中的发病风险。