Lnc000908通过内皮细胞-间质化调节心脏纤维化的机制研究

Cardiac fibrosis is the pathological process of the progress of various cardiovascular disease. Over-activation of EndMT plays a vital role in cardiac fibrosis. LncRNA is an important regulator of cardiac fibrosis. Which lncRNAs regulate EndMT to participate in cardiac fibrosis and how to regulate is a scientific problem to be solved urgently. Our previous work found a entirely new lncRNA-lnc000908 increased obviously in the fibrotic heart, its downstream target gene of EP4 was down regulated. Knock down of lnc000908 relieved cardiac fibrosis in rats, up regulated EP4 level. Studies have reported that activation of EP4 inhibit EndMT. Hence, we make a hypothesis that lnc000908 may inhibit EP4 transcription, promote EndMT, and increase cardiac fibrosis. This project intends to observe the dynamic expression of lnc000908 and EP4, and the effects of over-expression and knockout of lnc000908 on cardiac fibrosis; observe the effects of koncking down EP4 and koncking out lnc000908 on EndMT respectively to investigate whether lnc000908-EP4 pathway regulates cardiac fibrosis by EndMT; observe the effects of lnc000908 intervention on EP4 transcription to further explore the molecular mechanism of lnc000908 on EP4. The successful implementation of this project will provide an experimental basis for the early intervention of cardiac fibrosis with the lnc000908-EndMT pathway as a target.

心脏纤维化是心血管疾病进展的病理过程，过度激活EndMT在心脏纤维化起重要作用。LncRNA是心脏纤维化的重要调控因子。哪些lncRNA调节EndMT参与心脏纤维化及如何调节，是亟待解决的科学问题。前期工作发现纤维化心脏中一条新的lnc000908表达明显上调，下游EP4表达下调；敲低lnc000908减轻纤维化，上调EP4。据报道激活EP4抑制EndMT，由此提出lnc000908可能抑制EP4转录，促进EndMT，加重心脏纤维化。项目拟通过观察lnc000908及EP4动态表达，过表达及敲除lnc000908对心脏纤维化的影响；观察分别干预lnc000908及EP4对EndMT的影响，探讨其可能通过EndMT调控心脏纤维化；观察敲除lnc000908对EP4转录的影响，探讨其调控EP4的分子机制。该项目的成功实施将为以lnc000908-EndMT通路为靶点早期干预心脏纤维化提供依据。

心脏纤维化是心血管疾病进展的病理过程，EndMT的过度激活在心脏纤维化起重要作用。LncRNA是心脏纤维化的重要调控因子，且种类繁多。寻找具体的LncRNA和探究其如何调节EndMT参与心脏纤维化，是亟待解决的科学问题。本项目探讨了lnc000908 调节其下游靶基因 EP4 的表达参与 EndMT 的调节及心脏纤维化发生发展，提出了 EndMT 调节和心脏纤维化发病新的分子机制，发现lnc000908-EP4 可能通过 EndMT 调节心脏纤维化。SD大鼠通过皮下注射异丙索(5 mg/kg/d)连续7天，然后再饲养14天构建心衰模型，TGF-β被用来诱导大鼠心脏微血管内皮细胞(CMECs)产生EndMT现象，通过病毒低表达或过表达lnc000908，我们检测了心脏功能、纤维化、EndMT标记、lnc000908和EP4的表达水平。我们发现心脏微血管内皮富集的lnc000908在心脏纤维化中上调，沉默lnc000908通过抑制EndMT减少心肌纤维化，此外，Lnc000908通过抑制EP4受体促进EndMT。总之，以上结果表明，lnc000908的促纤维化作用是通过下调EP4介导的，lnc000908抑制EP4表达促进EndMT参与心脏纤维化，这说明lnc000908可能是治疗心脏纤维化的潜在治疗靶点。该项目资助发表SCI文章6篇，协助申请课题2项，其中国家自然科学基金面上项目1项，浙江省自然科学基金项目1项，培养研究生26名，其中9名已取得硕士学位，17名在读。