Ma Yu-Kun, Han Xin-Yi, Zan Shu-Huai, Liu Hui-Ting, Zhou Xue-Yan, Zhao Dan-Xue, Xing Rui, Zhao Peng
School of Basic Medicine, Qingdao University, Qingdao, 266000, China.
The First School of Clinical Medicine, Qingdao University, Qingdao, 266000, China.
Mol Cell Biochem. 2025 Apr;480(4):2487-2500. doi: 10.1007/s11010-024-05125-7. Epub 2024 Oct 7.
Cardiac remodeling is an end-stage manifestation of multiple cardiovascular diseases, and microRNAs are involved in a variety of posttranscriptional regulatory processes. miR-363-5p targeting Thrombospondin3 (THBS3) has been shown to play an important regulatory role in vascular endothelial cells, but the roles of these two in cardiac remodeling are unknown. Firstly, we established an in vivo model of cardiac remodeling by transverse aortic narrow (TAC), and then we stimulated a human cardiomyocyte cell line (AC16) and a human cardiac fibroblast cell line (HCF) using 1 μmol/L angiotensin II (Ang II) to establish an in vitro model of cardiac hypertrophy and an in vitro model of myocardial fibrosis, respectively. In all three of the above models, we found a significant decreasing trend of miR-363-5p, suggesting that it plays a key regulatory role in the occurrence and development of cardiac remodeling. Subsequently, overexpression of miR-363-5p significantly attenuated myocardial hypertrophy and myocardial fibrosis in vitro as evidenced by reduced the area of AC16, the cell viability of HCFs, the relative expression of the protein of fetal genes (ANP, BNP, β-MHC) and fibrosis marker (collagen I, collagen III, α-SMA), whereas inhibition of miR-363-5p expression showed the opposite trend. In addition, we also confirmed the targeted binding relationship between miR-363-5p and THBS3 by dual luciferase reporter gene assay, and the expression of THBS3 was directly inhibited by miR-363-5p. Moreover, overexpression of miR-363-5p with THBS3 simultaneously partially eliminated the delaying effect of miR-363-5p on myocardial hypertrophy and myocardial fibrosis in vitro. In conclusion, Overexpression of miR-363-5p attenuated the prohypertrophic and profibrotic effects of Ang II on AC16 and HCF by a mechanism related to the inhibition of THBS3 expression.
心脏重塑是多种心血管疾病的终末期表现,而微小RNA参与多种转录后调控过程。已证实靶向血小板反应蛋白3(THBS3)的miR-363-5p在血管内皮细胞中发挥重要调控作用,但二者在心脏重塑中的作用尚不清楚。首先,我们通过横向主动脉缩窄(TAC)建立了心脏重塑的体内模型,然后用1μmol/L血管紧张素II(Ang II)刺激人心肌细胞系(AC16)和人心脏成纤维细胞系(HCF),分别建立心脏肥大的体外模型和心肌纤维化的体外模型。在上述三种模型中,我们均发现miR-363-5p有显著下降趋势,提示其在心脏重塑的发生发展中起关键调控作用。随后,miR-363-5p的过表达显著减轻了体外心肌肥大和心肌纤维化,表现为AC16面积减小、HCF细胞活力降低、胎儿基因(ANP、BNP、β-MHC)蛋白和纤维化标志物(I型胶原、III型胶原、α-SMA)的相对表达降低,而抑制miR-363-5p表达则呈现相反趋势。此外,我们还通过双荧光素酶报告基因实验证实了miR-363-5p与THBS3之间的靶向结合关系,且miR-363-5p可直接抑制THBS3的表达。此外,miR-363-5p与THBS3同时过表达部分消除了miR-363-5p对体外心肌肥大和心肌纤维化的延缓作用。总之,miR-363-5p的过表达通过抑制THBS3表达的相关机制减弱了Ang II对AC16和HCF的促肥大和促纤维化作用。