冠心病患者血清外泌体微小核糖核酸-141-3p对内皮细胞损伤的影响研究  

Analysis of miR-141-3p in serum exosome from patients with coronary atherosclerosis disease and effects on vascular endothelial cells

作　　者：张帆[1] 张聪聪[1] 程乃萱 杜杰[1] 张海波[1] ZHANG Fan;ZHANG Congcong;CHENG Naixuan;DU Jie;ZHANG Haibo(Department of Cardiac Surgery,Beijing Anzhen Hospital,Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Diseases,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所心脏外科,100029

出　　处：《心肺血管病杂志》2021年第3期223-228,共6页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家自然科学基金项目(81871758)。

摘　　要：目的:检测冠状动脉粥样硬化性心脏病患者血清外泌体中微小核糖核酸-141-3p (miR-141-3p)的表达变化,并分析miR-141-3p对人血管内皮细胞损伤的影响。方法:收集冠状动脉粥样硬化性心脏病患者血清(n=8)和健康人对照(n=8),用外泌体提取试剂盒提取血清外泌体RNA,使用qRT-PCR的方法检测血清外泌体中miR-141-3p的表达变化;将人脐静脉内皮细胞(HUVECs)转染NC mimic或miR-141-3p mimic后使用ox-LDL诱导损伤,MTT法检测细胞活性,检测Annexin-V/PI染色结果评价细胞凋亡情况;生物信息学分析预测并验证miR-141-3p作用的靶基因。结果:与健康人的对照血清外泌体相比,冠状动脉粥样硬化患者的血清外泌体中miR-141-3p的表达显著增加(P<0.01)。在HUVECs中过表达miR-141-3p可抑制ox-LDL诱导的细胞凋亡,维持细胞活性(P<0.05);靶基因预测结果显示与细胞凋亡和增殖相关的基因PTEN可能是miR-141-3p的靶基因,进一步验证结果显示miR-141-3p过表达导致PTEN表达的下调(P<0.05)。结论:冠心病患者血清外泌体中miR-141-3p的表达上调,miR-141-3p可通过下调靶基因PTEN发挥抑制血管内皮细胞凋亡的保护性作用。miR-141-3p可能成为抗动脉粥样硬化治疗的新靶点。Objective: To examine the expression level of miR-141-3 p in serum exosome from patients with coronary atherosclerosis and confirm the effects of miR-141-3 p on vascular endothelial cells injury. Methods: Collected serum and extracted serum exosome from coronary atherosclerosis patients(n=8) and health controls(n=8), qRT-PCR was used to detect the expression levels of miR-141-3 p in serums;HUVECs were then transfected with NC mimic or miR-141-3 p mimic, and were treated with ox-LDL to induce cell apoptosis. Cell viability was detected by MTT, and apoptosis was evaluated by flow cytometry detection of Annexin-V/PI staining. The potential target genes of miR-141-3 p were predicted by TangetScan software, and the expression of target gene PTEN was detected by qRT-PCR. Results: Compared to serum exosome from healthy controls, the expression levels of miR-141-3 p were up-regulated significantly in the serum exosome from coronary atherosclerosis patients(P<0.01). After overexpressing miR-141-3 p in HUVECs by transfected with miR-141-3 p mimics, the apoptosis ratio of HUVECs was decreased and cell viability was maintained under ox-LDL stimulation conditions(P<0.05). Target gene prediction result showed that miR-141-3 p could combined with the 3’-UTR region of the PTEN gene, which was involved in apoptosis and proliferation. Further result showed that the expression of PTEN was significantly inhibited by the miR-141-3 p overexpression. Conclusions: The expression of miR-141-3 p in serum exosome from patients with coronary atherosclerosis disease was up-regulated, and miR-141-3 p could play a protective role in vascular endothelial cell by inhibiting the apoptosis associated gene PTEN. miR-141-3 p may become a new therapeutic target for atherosclerosis.

关 键 词：外泌体 微小核糖核酸-141-3p 人血管内皮细胞 细胞凋亡 动脉粥样硬化 

分 类 号：R54[医药卫生—心血管疾病]