Yang Jie, Xu Hong, Chen Kejie, Zheng Danni, Liu Shuang, Zhou Xia, Lin Yapeng, Cheng Hang, Luo Qin, Yang Min, Yan Xiaoyan, Hao Junli
Department of Neurology, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, China.
Department of Neurology, The First Affiliated Hospital of Chengdu Medical College, Chengdu, China.
Front Physiol. 2022 Jun 9;13:893102. doi: 10.3389/fphys.2022.893102. eCollection 2022.
The interaction between platelets and vascular endothelial cells plays a pivotal role in the pathophysiology of acute ischemic stroke (AIS), especially in atherosclerosis formation. However, the underlying mechanism is not entirely clear. The aim of this study was to elucidate the role of platelets-derived miRNA in the development of atherosclerosis and AIS. We evaluated the miRNA expression profiles of serum microvesicles (MV) in five AIS patients and five healthy controls using RNA-seq, and then measured the levels of selected platelets derived miRNAs by qRT-PCR. miR-200a-3p expression in the serum MV and platelets had increased to 1.41 ( < 0.05) and 3.29 times ( < 0.001), respectively, in AIS patients compared with healthy controls, and was modified by severity of AIS. We transferred Cy5-miR-200a-3p into platelets, collected and identified platelets-derived MV (PMVs). Then, the gene expression of p38 MAPK/c-Jun pathway was analyzed using both miR-200a-3p gain- and loss-of-function experiments and PMVs coincubation with HUVEC. The results showed that activated platelets remotely modulated endothelins 1 (ET-1) and vascular endothelial growth factor A (VEGFA) levels in HUVEC through the release of miR-200a-3p-containing PMVs targeting MAPK14. The results of ROC analyses showed that combination of platelet miR-200a-3p, serum ET-1 and VEGFA levels had an AUC of 0.817, a sensitivity of 70%, and a specificity of 89%. Our results presented new evidence that activated platelets could remotely modulate ET-1 and VEGFA expression in HUVEC via releasing miR-200a-3p-enriched PMVs, which provides a potential miRNA-based predictive biomarker and therapeutic strategy for atherosclerosis and AIS.
血小板与血管内皮细胞之间的相互作用在急性缺血性卒中（AIS）的病理生理学中起着关键作用，尤其是在动脉粥样硬化形成过程中。然而，其潜在机制尚不完全清楚。本研究的目的是阐明血小板衍生的微小RNA在动脉粥样硬化和AIS发展中的作用。我们使用RNA测序评估了5例AIS患者和5例健康对照血清微泡（MV）中的微小RNA表达谱，然后通过qRT-PCR测量了所选血小板衍生微小RNA的水平。与健康对照相比，AIS患者血清MV和血小板中miR-200a-3p的表达分别增加到1.41倍（P<0.05）和3.29倍（P<0.001），并且受AIS严重程度的影响。我们将Cy5-miR-200a-3p转入血小板，收集并鉴定血小板衍生的MV（PMV）。然后，使用miR-200a-3p功能获得和功能丧失实验以及PMV与HUVEC共孵育来分析p38 MAPK/c-Jun途径的基因表达。结果表明，活化的血小板通过释放靶向MAPK14的含miR-200a-3p的PMV远程调节HUVEC中内皮素1（ET-1）和血管内皮生长因子A（VEGFA）的水平。ROC分析结果显示，血小板miR-200a-3p、血清ET-1和VEGFA水平联合检测的曲线下面积（AUC）为0.817，灵敏度为70%，特异性为89%。我们的结果提供了新的证据，表明活化的血小板可通过释放富含miR-200a-3p的PMV远程调节HUVEC中ET-1和VEGFA的表达，这为动脉粥样硬化和AIS提供了一种基于微小RNA的潜在预测生物标志物和治疗策略。
