Suppressing circ_0008494 inhibits HSCs activation by regulating the miR-185-3p/Col1a1 axis.
Suppressing circ_0008494 inhibits HSCs activation by regulating the miR-185-3p/Col1a1 axis.
复制标题
抑制 circ_0008494 通过调节 miR-185-3p/Col1a1 轴抑制 HSC 激活
DOI：
10.3389/fphar.2022.1050093
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发表时间：
2022
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
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作者：
背景：肝纤维化的特征是肝星状细胞(HSCs)活化和细胞外基质成分广泛沉积，尤其是胶原蛋白。然而，有效的抗肝纤维化治疗仍然缺乏。最近，环状RNA(CircRNAs)被发现是一种新型的HF调节因子。方法：用测序方法筛选CircRNAs图谱，用荧光原位杂交法确定CIRC_0008494在人心衰组织中的定位。结果预测采用生物信息学分析，双荧光素酶报告结合AgO-RIP和生物素偶联miRNA捕获实验确定miR185-3p/I型胶原α1链(Col1a1)为CIRC_0008494的靶点。构建了稳定的CIRC_0008494干扰人HSCs细胞系，并用于研究CIRC_0008494/miR-185-3p/Col1a1轴的调控机制。结果：CIRC_0008494在人肝纤维化组织中大量且显著过表达，定位于肝星状细胞胞浆。双重荧光素酶报告、AgO-RIP和生物素偶联的miRNA捕获实验共同证实了CIRC_0008494是miR-185-3p的海绵。细胞功能实验和挽救实验表明，抑制CIRC_0008494可通过抑制miR-185-3p抑制HSC的激活、增殖、迁移和促进其凋亡。特别是，HF指示剂Col1a1被证实为miR-185-3p的直接靶点，而CIRC_0008494的抑制通过释放miR-185-3p来抑制Col1a1的表达。结论：CIRC185p/Col1a1轴基因敲除可抑制HSCs活化。CIRC_0008494有望成为治疗心力衰竭的靶点。
Background: Hepatic fibrosis (HF) is characterized by activation of hepatic stellate cells (HSCs) and extensive deposition of extracellular matrix components, especially collagens. However, effective antifibrotic therapies are still lacking. Recently, circular RNAs (circRNAs) have been identified as novel regulators of HF. Methods: circRNAs profile was screened by RNA sequencing and the location of circ_0008494 was confirmed by fluorescence in situ hybridization assay in human HF tissues. Bioinformatics analysis was used for result prediction and dual-luciferase reporter, together with AGO-RIP and biotin-coupled miRNA capture assays, were used to determine miR-185-3p/collagen type I alpha 1 chain (Col1a1) as the target of circ_0008494. A stable circ_0008494-interfering human HSCs cell line was constructed and used to determine the regulatory mechanism of circ_0008494/miR-185-3p/Col1a1 axis. Results: circ_0008494 was abundantly and significantly over-expressed in human HF tissues and located at the cytoplasm of HSCs. Together, dual-luciferase reporter, AGO-RIP and biotin-coupled miRNA capture assays confirmed that circ_0008494 acted as a sponge of miR-185-3p. Cell functional experiments and rescue assays demonstrated suppressing circ_0008494 could inhibit activation, proliferation, migration of HSCs and promote their apoptosis through miR-185-3p. In particular, the HF indicator, Col1a1, was validated as the direct target of miR-185-3p and the suppression of circ_0008494 inhibited the expression of Col1a1 by releasing miR-185-3p. Conclusion: Knocking down circ_0008494 inhibited HSCs activation through the miR-185-3p/Col1a1 axis. circ_0008494 could be a promising treatment target for HF.