Liraglutide improves atherosclerosis by regulating long non-coding RNA RMRP/miR-1 28-1-5P/Gadd45g axis
Liraglutide improves atherosclerosis by regulating long non-coding RNA RMRP/miR-1 28-1-5P/Gadd45g axis
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DOI：
10.26355/eurrev_202003_20545
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发表时间：
2020-01-01
影响因子：
3.3
通讯作者：
Shi, F-W
中科院分区：
文献类型：
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作者：
An, J-H;Chen, Z-Y;Shi, F-W
目的：近年来的研究表明，长链非编码RNA参与了动脉粥样硬化的形成，而动脉粥样硬化是冠心病的病理基础。材料与方法：采用qPCR方法检测IL-6、IL-8、RMRP和miR-128-1 - 5 P在冠状动脉粥样硬化和人血管平滑肌细胞中的表达。荧光素酶报告基因分析证实了RMRP与miR-128-1- 5 P和miR-128-1- 5 P与Gadd 45 g对HEK 293 T的直接靶向作用。结果：RMRP和Gadd 45 g蛋白在冠状动脉粥样硬化和人血管平滑肌细胞中表达上调，而miR-128-1- 5 P表达下调。RMRP下调显著抑制ox-LDL处理后人血管平滑肌细胞IL-6、IL-8及凋亡相关蛋白的表达。此外，生物信息学分析和荧光素酶报告实验证实RMRP是miR-128-1- 5 P的直接靶点。miR-128-1- 5 P抑制剂能明显抑制ox-LDL诱导的IL-6、IL-8和凋亡相关蛋白RMRP-si的作用，提示RMRP对miR-128-1- 5 P在冠状动脉粥样硬化中起负向和直接调控作用。更重要的是，RMRP沉默增加了人血管平滑肌细胞中Gadd 45 g蛋白的水平。当miR-128上调时，发现了相同的结果。同时，Gadd 45 g-si极大地逆转了ox-LDL和Luciferase处理人血管平滑肌细胞后IL-6、IL-8和凋亡相关蛋白诱导的miR-128-1- 5 P抑制剂的作用结果。实验结果表明，Gadd 45 g是miR-128-1- 5 P的直接靶点，提示Gadd 45 g在冠状动脉粥样硬化中负向直接调控miR-128-1- 5 P。利拉鲁肽还能明显抑制冠状动脉粥样硬化中IL-6、IL-8及凋亡相关蛋白的表达。结论：利拉鲁肽可通过调节RMRP/miR-128 - 1 - 5 P/Gadd 45 g信号通路抑制冠状动脉粥样硬化中炎性细胞因子和凋亡相关蛋白的表达，为冠状动脉粥样硬化的治疗提供了新的潜在策略。
OBJECTIVE: Recent studies indicated that long non-coding RNA is involved in the formation of atherosclerosis, which is the pathological basis of coronary heart disease. Here, we reported the function and regulatory mechanism of RMRP in coronary atherosclerosis.MATERIALS AND METHODS: qPCR was used to investigate the expression of IL-6, IL-8, RMRP, and miR-128-1 -5P in coronary atherosclerosis and human vascular smooth muscle cells. Luciferase reporter assay confirmed the direct target effect of RMRP with miR-128-1-5P and miR-128-1-5P with Gadd45g on HEK293T. Western blot was used to detect protein expression in coronary atherosclerosis and human vascular smooth muscle cells.RESULTS: RMRP expression and Gadd45g protein level were up-regulated in coronary atherosclerosis and human vascular smooth muscle cells, while miR-128-1-5P was down-regulated. RMRP downregulation remarkably inhibited the expression of IL-6, IL-8, and apoptosis related protein in human vascular smooth muscle cells after ox-LDL treatment. In addition, bioinformatics analysis and Luciferase report experiments confirmed that RMRP was the direct target of miR-128-1-5P. Moreover, miR-128-1-5P inhibitor reserved evidently the effect of IL-6, IL-8, and apoptosis related protein induced RMRP-si after treatment of human vascular smooth muscle cells with ox-LDL, implying RMRP negatively and directly regulated miR-128-1-5P in coronary atherosclerosis. More importantly, RMRP silencing increased Gadd45g protein level in human vascular smooth muscle cells. The same results were found when miR-128 was upregulated. Meanwhile, Gadd45g-si extremely reversed the result of IL-6, IL-8, and apoptosis related protein induced miR-128-1-5P inhibitor after treatment of human vascular smooth muscle cells with ox-LDL and Luciferase report experiments showed that Gadd45g was a direct target of miR-128-1-5P, implying Gadd45g negatively and directly regulated miR-128-1-5P in coronary atherosclerosis. Furthermore, liraglutide restrained evidently the expression of IL-6, IL-8, and apoptosis related protein in coronary atherosclerosis. After all, these results showed that liraglutide could regulate RMRP/miR-128-1-5P/Gadd45g signal pathway to improve coronary atherosclerosis.CONCLUSIONS: Liraglutide could curb the expression of inflammatory cytokines and apoptosis related protein in coronary atherosclerosis by regulating RMRP/miR-128-1-5P/Gadd45g signaling pathway, providing a new potential strategy for the treatment of coronary atherosclerosis.