TGFβ2 相关lncRNA--TGFβ2RLR在去门静脉化诱导肝再生中的作用研究

Liver has strong ability of regeneration which would be triggered mediated by the blood stream dynamics of portal vein supply. The applicant has focused on the field of liver regeneration in clinical and scientific research works for long time, and finished many cases of changing portal vein blood distribution to improve liver regeneration in medical treatment. We also have built the model of rat liver regeneration/atrophy by changing hepatic portal vein blood distribution and investigated the change of long chain non-coding RNA (lncRNA) of rat liver tissue in previous research. We found there was significant differences in expression of lncRNAs between hyperplasia and atrophy rat liver tissue. And the results indicated that lncRNAs may play an important role in the process of liver regeneration. The results of bioinformatics analysis and validation showed that lncRNA was strongly correlated with TGF β2, which was significantly up-regulated at the initiation stage of rat liver regeneration and had a positive correlation with the end of liver regeneration. The aim of this project is to discuss the mechanism of lncRNA-TGF β2RLR in the process of liver regeneration, and to investigate the effect and feasibility about using lncRNA-TGF β2RLR targeted on liver regeneration to treat the patients with liver disease.

肝脏具有强大的再生能力，门静脉血流对启动、维系肝再生发挥关键性的体液及血流动力学介导调控机制。申请人长期关注于肝再生临床及科研工作，利用改变肝脏门静脉血流分布、促进肝再生开展了大量的医疗实践工作。结合临床问题设计了去门静脉化大鼠肝再生/萎缩自身对照模型，进行大鼠肝组织长链非编码RNA芯片的制备、筛选及验证等前期研究工作发现，大鼠两侧增生/萎缩肝组织有大量的差异lncRNA表达，提示lncRNA在肝再生过程中发挥着重要作用。通过生物信息学分析及验证结果表明，lncRNA 与TGF β2具有强烈相关性，在大鼠肝再生启动阶段即显著上调，并与肝再生状态的维系、结束存在正相关性。本项目拟在前期工作基础上结合大鼠肝再生体内、体外模型，探讨、验证lncRNA--TGF β2RLR在肝再生过程中的作用机制和分子调控机制，并探讨在临床肝病治疗中应用靶向干预lncRNA以改善肝再生的可行性。

肝脏体液及血流动力学的变化尤其是肝脏门静脉血流供应的变化，在肝再生过程中发挥关键性的作用。为了揭示去门静脉化加速大鼠肝再生的机制，我们构建了大鼠门静脉结扎（PVL）模型，对PVL后0、1、7和14天的非结扎肝叶进行了全面的转录组分析，鉴定了大量差异表达的长链非编码RNA（lncRNA）及编码基因，通过差异基因进行加权基因共表达网络分析(WGCNA)构建了10个模块，鉴定了与肝再生相关的几个关键生物学过程，如补体激活、IL-6生成、Wnt信号通路、自噬等。16个mRNAs (Notch1、Grb2、IL-4、Cops4、Stxbp1、Khdrbs2、Hdac2、Gnb3、Gng10、Tlr2、Sod1、Gosr2、Rbbp5、Map3k3、Golga2、Rev3l)和10个lncRNAs (BC092620、AB190508、EF076772、BC088302、BC158675、BC100646、BC089934、L20987、BC091187、M23890)被确定为关键基因，其中BC092620在肝细胞增殖及肝再生中的功能正在研究中。.为了在不同肝再生动物模型中鉴定功能性lncRNA，我们构建了小鼠2/3部分肝切除模型。LncRNA-TGFβ2RLR与SNHG12在2/3 PH后的不同时间点均表达上调，在正常肝细胞系中，不同浓度的肝细胞生长因子处理后，SNHG12在不同时间点也均呈显著上调。功能上，尽管TGFβ2RLR对肝细胞增殖的促进作用并不明显，但SNHG12过表达在体内外均能促进肝细胞增殖，过表达SNHG12通过激活Wnt/β-catenin通路，促进了小鼠肝再生。本研究显示，通过人为干预门静脉血流中特定lncRNA的表达，可有效提高肝脏再生能力，能够增加未来残肝体积，提高大肝癌的可切除性。