CircRNA97介导miR-146b-KLF4在肝纤维化逆转中的作用机制研究

MicroRNA（miRNA) plays an important role in the development of hepatic fibrosis by modulating the activation of hepatic stellate cells (HSCs). Circular RNA (circRNA) involves in regulating RNA network by miRNA as a spongelike role, intensive study of the underlying mechanism has become one of research direction to explore effective reversal strategies for hepatic fibrosis.In the previous study, we have conducted the role of miRNA-146b-KLF4 in the activation of HSCs. Using circRNA predictive software and high-through sequencing technology, we found that circRNA97 could be conjunct with miR-146b, and might affect the function of miR-146b specifically. We here infer that circRNA97 could regulate the HSCs activation by miR-146b-KLF4 signaling cascade pathway. To verify this hypothesis and further explore the mechanism of circRNA97 in hepatic fibrosis recover,from the level of the molecules, cells, tissues and animals,molecular and cell biological technology will be employed.This subject will reveal a novel mechanism in HSCs activation by circRNA, and provide new sight to explore the reversal of hepatic fibrosis.

小分子RNA（miRNA）调控肝星状细胞（HSCs）的活化在肝纤维化发展中具有重要作用。环状RNA（circRNA）通过miRNA参与调控RNA-RNA网络发挥海绵样作用，深入探索两者之间的调控机制成为逆转肝纤维化的研究方向之一。我们在前期工作中已经发现miR-146b-KLF4途径在HSCs活化中的作用，并运用circRNA靶点miRNA预测软件和高通量测序筛选获得靶向调控miR-146b的circRNA97。据此我们提出假说circRNA97通过miR-146b-KLF4信号通路调控HSCs的活化。本项目将应用分子和细胞生物学技术，从分子、细胞、组织和动物整体水平验证circRNA97调控miR-146b-KLF4的信号通路，探讨circRNA97在肝纤维化逆转过程中的作用机制。本研究将从circRNA这个新视点揭示HSCs活化的机制奠定基础，为肝纤维化逆转提新的思路。

小分子RNA（miRNA）调控肝星状细胞（HSCs）的活化在肝纤维化发展中具有重要作用。环状RNA（circRNA）通过调控miRNA参与肝星状细胞（HSCs）的活化过程从而影响肝纤维化的发生机制是本项目的研究重点。本研究发现circRNA97过表达能显著增加肝纤维化标志物的表达，miR-146b-5p在肝纤维化大鼠和活化的HSC-T6中均显著升高，过表达HIPK1能有效逆转miR-146b-5p对肝纤维化的促进作用，说明miR-146b-5p可作为肝纤维化的潜在标志物和治疗靶标。另外，circRNA97通过竞争性结合miR-128-3p提高Bcl-2/Bax的表达，增加a-SMA、TNF-a，TGF-β等的表达，促进HSC-T6的活化从而诱导肝纤维化，而过度的TGF-β活性水平可通过COL1A1形成TGF-β1水平的反馈回路，准确控制TGF-β可溶性受体表达抑制肝纤维化且不会触发自身免疫反应。本项目证实了circRNA97可介导miRNA调控HSCs的活化机制参与肝纤维化的发生，为进一步逆转肝纤维化的发生和新型靶向药物研发提供了新的思路和方法。