【罂粟摘要】抑制circHIPK3可通过海绵转染miR-29b-3p来抑制血管紧张素II诱导的心肌纤维化

抑制circHIPK3可通过海绵转染miR-29b-3p来抑制血管紧张素II诱导的心肌纤维化

翻译：安丽    编辑：佟睿    审校：曹莹

贵州医科大学高鸿教授课题组

背景

环状RNA (circRNAs)是心脏发育和疾病强有力的调节因子。然而，描述环状RNA介导的心脏成纤维细胞(CFs)生物学调控及其在心脏纤维化中的作用的详细研究仍然有限。

方法

使用PCR和Sanger测序检测circHIPK3在CFs中的表达。 采用Transwell迁移法和免疫荧光染色法检测circHIPK3在体外CFs中的作用。采用生物信息学分析、双荧光素酶活性分析、RNA免疫沉淀和荧光原位杂交实验研究circHIPK3介导的心肌纤维化的机制。采用超声心动图分析、Sirius Red染色和免疫荧光染色研究circHIPK3在血管紧张素II (Ang II)诱导的体内心肌纤维化中的作用。

结果

Ang II治疗后CFs和心脏组织中circHIPK3表达明显增加。circHIPK3沉默可抑制体外Ang II诱导的CFs增殖、迁移和a-SMA表达水平的上调。circHIPK3作为miR-29b-3p海绵，过表达circHIPK3有效逆转了miR-29b-3p诱导的CFs增殖和迁移抑制，改变了miR-29b-3p靶向基因(a- SMA、COL1A1、COL3A1)的体外表达水平。circHIPK3沉默和miR-29b-3p过表达联合对体内心脏纤维化抑制的作用强于circHIPK3沉默或miR-29b-3p过表达。

结论

我们的数据表明circHIPK3作为miR-29b-3p海绵调节CFs增殖、迁移和心脏纤维化的发展，揭示了一个潜在的预防Ang II诱导的心脏纤维化的新靶点。

Inhibition of circHIPK3 prevents angiotensin II-induced cardiac fibrosis by sponging miR-29b-3p

Abstract

Background: Circular RNAs (circRNAs) are emerging as powerful regulators of cardiac development and disease.Nevertheless, detailed studies describing circRNA-mediated regulation of cardiac fibroblasts (CFs) biology and their role in cardiac fibrosis remain limited.

Methods: PCR and Sanger sequencing were performed to identify the expression of circHIPK3 in CFs. Educorporation assays, Transwell migration assays, and immunofluorescence staining assays were conducted to detect the function of circHIPK3 in CFs in vitro. Bioinformatics analysis, dual luciferase activity assays,RNA immunoprecipitation, and fluorescent in situ hybridization experiments were conducted to investigate the mechanism of circHIPK3-mediated cardiac fibrosis. Echocardiographic analysis, Sirius Red staining and immunofluorescence staining were performed to investigate the function of circHIPK3 in angiotensin II(Ang II) induced cardiac fibrosis in vivo.

Results: circHIPK3 expression markedly increased in CFs and heart tissues after the treatment of Ang II. circHIPK3 silencing attenuates CFs proliferation, migration and the upregulation of a-SMA expression levels induced by Ang II in vitro. circHIPK3 acted as a miR-29b-3p sponge and overexpression of circHIPK3 effectively reverses miR-29b-3p-induced inhibition of CFs proliferation and migration and alters the expression levels of miR-29b-3p targeting genes (a-SMA, COL1A1, COL3A1) in vitro. Combination of circHIPK3 silencing and miR-29b-3p overexpression had a stronger effect on cardiac fibrosis suppression in vivo than did circHIPK3 silencing or miR-29b-3p overexpression alone.

Conclusions: Our data suggest that circHIPK3 serves as a miR-29b-3p sponge to regulate CF proliferation, migration and development of cardiac fibrosis, revealing a potential new target for the prevention of Ang II-induced cardiac fibrosis.