circRNATUBD1作为miR-146a-5p海绵参与调控TLR4信号通路影响放射性肝病发生的分子机制研究

miR-146a-5p负向调控肝星状细胞(HSC)TLR4通路，影响放射性肝病(RILD)发生发展，但其上游调控机制仍不清楚。前期实验显示，与miR-146a-5p有结合位点的环状RNA TUBD1(circTUBD1，hsa_circ_0044897)在接受照射的HSC中表达明显升高，且其表达变化影响TLR4通路分子的表达，故我们推测circTUBD1可能通过竞争性结合miR-146a-5p参与调控TLR4信号通路，从而影响RILD的发生发展。本项目拟采用体内外实验与临床验证相结合，应用RAP-qRT-PCR、AGO2免疫共沉淀、基因过表达或干扰等实验技术，探讨circTUBD1作为miR-146a-5p海绵，参与竞争性内源RNA调控模式，影响肝星状细胞功能、TLR4信号通路、肝微环境和肝细胞放射敏感性的机制，为RILD发病机制的研究开拓思路、提供防治策略。

MiR-146a-5p negatively regulates TLR4 pathway in hepatic stellate cell (HSC) to influence the development of radiation-induced liver disease (RILD), but the upstream regulation mechanism remains unclear. Preliminary study reveals that circRNA TUBD1(circTUBD1，hsa_circ_0044897) with miR-146a-5p binding site is significantly up-regulated and correlated with the changes of TLR4 pathway molecules in irradiated HSC, thus we speculate that circTUBD1 may be involved in the regulation of TLR4 signaling pathway to participate in the development of RILD through competitive binding of miR-146a-5p. This project is intended to use the in-vitro and in-vivo animal experiments in combination with the clinical validation to investigate the mechanism that circTUBD1 serving as the miR-146a-5p sponge involved in competing endogenous RNA regulation pattern to affect the function of HSC, TLR4 signaling pathway, hepatic micro-environment and radiation sensitivity of hepatocyte, by use of RNA antisense purification-quantitative real-time PCR, immuneprecipitation of AGO2, gene overexpression or interference. This project may offer a new perspective to understand the mechanism as well as providing the preventative and therapeutic strategies for RILD.