miR-210通过调节巨噬细胞对免疫性肝损伤的影响及机制研究

Immune-mediated liver injury is a seriously social public problem in our country and even in the world. Regulating immune response is an important to control immunological liver injury and indispensable to retard the progress of liver disease. Recent studies have shown that macrophages are essential in development and progression of immunological liver injury. Our previous results found that macrophages accelerate immune-mediated liver injury. And miR-210 expression was significantly up-regulated in the mouse serum and liver tissue during immunological liver injury. In addition, miR-210 inhibits macrophage to secrete inflammatory cytokines. Above results suggest that miR-210 may play a protective role in immune liver injury by negatively regulating macrophage function. Therefore, to prove the hypothesis in this project, we will investigate the effect and mechanism of miR-210 to immunological liver injury with immunofluorescence, flow cytometry, dual luciferase reporter assay, TUNEL and ELISA via cell and animal models. This study will further elucidate the mechanism of immunological liver injury and provide a theoretical basis to estimate the status and process of immunological liver injury for miR-210.

免疫性肝损伤已成为严重威胁我国甚至全球人民健康的社会公共问题，调节免疫反应对于控制肝损伤尤为关键，是减缓其进程的重要环节。近年研究显示，巨噬细胞在免疫性肝损伤发生发展过程中至关重要，申请者前期研究发现：巨噬细胞促进免疫性肝损伤；免疫性肝损伤时，小鼠血清、肝组织和肝单个核细胞中miR-210表达都显著上调；miR-210抑制巨噬细胞炎性细胞因子分泌。前期结果推测，miR-210可能通过抑制巨噬细胞功能对免疫性肝损伤起到一定的保护作用。为论证该假说，本项目拟以细胞模型和动物模型为研究对象，在过表达和功能缺失两个层面，采用免疫荧光、流式细胞术、双荧光报告基因、TUNEL以及ELISA等技术，研究和探讨miR-210通过调节巨噬细胞对免疫性肝损伤的影响和机制。本研究将进一步阐明免疫性肝损伤的发生机制，并为miR-210作为评估免疫性肝损伤进程提供依据。

乙型肝炎病毒感染是严重威胁人类健康的重大疾病之一。HBV感染本身对肝细胞所产生的病理效应是微不足道的，它所引起的相关肝脏疾病的严重程度和结果主要是由机体的固有免疫反应和适应性免疫反应对抗HBV病毒相关抗原而引起的免疫性损伤所致，被认为是一种免疫性肝损伤疾病。本研究小组前期研究报道，巨噬细胞在HBV诱导的免疫性肝脏损伤的发病机制中发挥着重要作用。但是，HBV感染时，巨噬细胞的活化机制还未完全阐明？通过该项目的实施，本研究小组不但证实了HBV感染对巨噬细胞的激活作用，而且探讨了HBeAg对巨噬细胞miRNAs表达的影响和机制，并分析了miR-155和-210与HBV感染患者多种临床指标的相关性，得出如下结论：1）HBV感染促进了巨噬细胞活化。2）HBeAg促进了巨噬细胞miR-155的表达。3）NF-κB和PI3K信号通路在HBeAg诱导的巨噬细胞miR-155表达中发挥着重要的作用。4）HBeAg诱导的miR-155通过靶向于BCL-6、SHIP-1和SOCS-1促进其诱导的炎症反应。5）急性乙肝患者单个核细胞中miR-155的表达显著增强，且与血清中的ALT、AST和HBeAg含量密切相关。6）与正常对照组相比，HBV感染患者外周血单核细胞和血清中miR‑210的表达显著下降，并且与患者血清ALT和AST呈负相关。7）体外细胞培养和抗原刺激实验证实，在不同类型的巨噬细胞中，miR-210表达不受HBV相关抗原影响。8）CHB患者的血清显著下调了健康对照者外周血单核细胞miR-210的表达。该项目研究结果为miR-155和-210作为诊断、治疗和监测乙肝患者的肝脏免疫状态提供了重要依据，具有潜在的实际应用价值。