Qin Hao, Hu Long-Long, Wang Wen-Jun, Yu Zuo-Zhong, Chen Yang, Zhao Yuan-Bin, Liao Yan-Hui, Zhang Wei-Lin, Yang Ren-Qiang
Department of Cardiovascular Medicine, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, People's Republic of China.
Department of Respiratory Diseases, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, People's Republic of China.
Exp Gerontol. 2023 Dec;184:112330. doi: 10.1016/j.exger.2023.112330. Epub 2023 Nov 16.
Hyperhomocysteinemia (HHcy) is an independent risk factor of atherosclerosis (AS). Some reports have shown that homocysteine (Hcy) could accelerate the development of AS by promoting endothelial cell senescence. miRNAs were widely involved in the pathophysiology of HHcy. However, few studies have focused on the changes of miRNA-mRNA networks in the artery of HHcy patients. For this reason, RNA-sequencing was adopted to investigate the expression of miRNA and mRNA in HHcy model mouse arteries. We found that the expression of 216 mRNAs and 48 miRNAs were significantly changed. Using TargetScan and miRDB web tools, 29 miRNA-mRNA pairs were predicted. Notably, miR-20b-5p and FJX1 shared the highest predicted score in TargetScan, and further study indicated that the miR-20b-5p inhibitor significantly upregulated the FJX1 expression in HHcy human umbilical vein endothelial cells (HUVECs) model. PPI analysis revealed an important sub-network which was centered on CDK1. Gene ontology (GO) enrichment analysis showed that HHcy had a significant effect on cell cycle. Further experiments found that Hcy management increased reactive oxygen species (ROS) generation, the activity of senescence associated β-galactosidase (SA-β-gal) and the protein expression of p16 and p21 in HUVECs, which were rescued by miR-20b-5p inhibitor. In general, our research indicated the important role of miR-20b-5p in HHcy-related endothelial cell senescence.
高同型半胱氨酸血症(HHcy)是动脉粥样硬化(AS)的独立危险因素。一些报告表明,同型半胱氨酸(Hcy)可通过促进内皮细胞衰老加速AS的发展。微小RNA(miRNA)广泛参与HHcy的病理生理学过程。然而,很少有研究关注HHcy患者动脉中miRNA-mRNA网络的变化。因此,采用RNA测序技术研究HHcy模型小鼠动脉中miRNA和mRNA的表达。我们发现216种mRNA和48种miRNA的表达发生了显著变化。使用TargetScan和miRDB网络工具,预测了29对miRNA-mRNA。值得注意的是,miR-20b-5p和FJX1在TargetScan中的预测得分最高,进一步研究表明,miR-20b-5p抑制剂显著上调了HHcy人脐静脉内皮细胞(HUVECs)模型中FJX1的表达。蛋白质-蛋白质相互作用(PPI)分析揭示了一个以细胞周期蛋白依赖性激酶1(CDK1)为中心的重要子网。基因本体(GO)富集分析表明,HHcy对细胞周期有显著影响。进一步实验发现,Hcy处理增加了HUVECs中活性氧(ROS)的生成、衰老相关β-半乳糖苷酶(SA-β-gal)的活性以及p16和p21蛋白的表达,而miR-20b-5p抑制剂可使其恢复。总体而言,我们的研究表明miR-20b-5p在HHcy相关的内皮细胞衰老中起重要作用。