Ligustrazine prevents coronary microcirculation dysfunction in rats via suppression of miR-34a-5p and promotion of Sirt1
Ligustrazine prevents coronary microcirculation dysfunction in rats via suppression of miR-34a-5p and promotion of Sirt1
复制标题
川芎嗪通过抑制 miR-34a-5p 和促进 Sirt1 预防大鼠冠状动脉微循环功能障碍
DOI：
10.1016/j.ejphar.2022.175150
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发表时间：
2022
影响因子：
5
通讯作者：
Ma XiaoJuan
中科院分区：
文献类型：
--
作者：
Gao Jie;Ren Junguo;Ma Xin;Zhang Ying;Song Lei;Liu Jianxun;Shi DaZhuo;Ma XiaoJuan
冠状动脉微栓塞导致冠状动脉微血管功能障碍（CMD）， miR-34a-5p可能在其中起关键作用。据报道，川芎嗪可以改善CMD。本研究旨在探讨miR-34a-5p/Sirt1通路在CMD中的作用，并探讨川芎嗪的潜在机制。方法左心室注射月桂酸钠诱导大鼠冠状动脉微栓塞。通过HE染色和血流动力学检查观察CME的形成和心功能。RT-qPCR和western blot检测miR-34a-5p、Sirt1及下游蛋白的表达。双荧光素酶报告基因（DLR）检测证实miR-34a-5p与Sirt1之间的联系。采用ELISA法检测各组血管内皮功能障碍、血小板活化及炎症指标。结果CME大鼠出现明显的CME和心功能障碍，miR-34a-5p上调，Sirt1下调。过表达miR-34a-5p加重，而沉默miR-34a-5p抑制CME的形成。DLR检测证实miR-34a-5p直接抑制Sirt1 mRNA表达。川芎嗪预处理抑制miR-34a-5p，促进Sirt1表达，减轻内皮功能障碍，抑制血小板活化和炎症，从而减少CME。过表达miR-34a-5p降低了川芎嗪的积极作用；而沉默miR-34a-5p后，川芎嗪不能进一步促进Sirt1表达和抑制CME的形成。结论mir -34a-5p通过抑制Sirt1表达参与CMD的发生。川芎嗪通过抑制miR-34a-5p和促进Sirt1，发挥内皮保护、抗血小板和抗炎作用，预防CMD。
IntroductionThe coronary microembolization contributes to coronary microvascular dysfunction (CMD), in which miR-34a-5p may play a critical role. Ligustrazine has been reported to improve CMD. The present study was designed to discuss the role of miR-34a-5p/Sirt1 pathway in CMD and explore the underlying mechanism of ligustrazine.MethodsCoronary microembolization (CME) was induced by left ventricle injection of sodium laurate in rats. CME formation and cardiac function were examined by HE staining and hemodynamic tests to evaluate CMD. The expressions of miR-34a-5p, Sirt1 and the downstream proteins were detected by RT-qPCR and western blot. Dual-luciferase reporter (DLR) assay was performed to confirm the connection between miR-34a-5p and Sirt1. The blood markers of endothelial dysfunction, platelet activation and inflammation were examined with ELISA.ResultsOvert CME and cardiac dysfunction as well as up-regulated miR-34a-5p and down-regulated Sirt1 were observed in CME rats. Overexpressing miR-34a-5p aggravated while silencing miR-34a-5p inhibited CME formation. DLR assay confirmed that miR-34a-5p directly inhibited Sirt1 mRNA expression. Ligustrazine pretreatment suppressed miR-34a-5p and promoted Sirt1 expression, which alleviated endothelial dysfunction, inhibited platelet activation and inflammation, and in turn reduced CME. Overexpressing miR-34a-5p diminished the positive effects of ligustrazine; while after silencing miR-34a-5p, ligustrazine failed to further promote Sirt1 expression and inhibit CME formation.ConclusionMiR-34a-5p contributes to CMD by inhibiting Sirt1 expression. Ligustrazine exerts endothelial-protective, anti-platelet and anti-inflammatory effects to prevent CMD via suppressing miR-34a-5p and promoting Sirt1.