长链非编码RNA通过调控巨噬细胞分化参与颅内动脉瘤破裂的机制研究

The imbalances of M1/M2 macrophages polarity differentiation was the mainly reason for the rupture of intracranial aneurysm (IA) and miRNAs played an important role in the process of macrophages polarity differentiation. Existed studies have found that long non-coding RNA (lncRNAs) could competitive with endogenous mRNAs as (ceRNA) that effectively regulated the activity of miRNAs. But whether this mechanism underlying the process of macrophage polarity differentiation process, which resulted in inducing the rupture of intracranial aneurysms, still uncertain. Our previous study has confirmed the distinct lncRNAs and miRNAs among the process of the rupture of human intracranial aneurysm and macrophage polarization differentiation by gene chip technology. Proposed on this basis, the further research would focus on: 1) intracranial aneurysm model of rats would constructed after knockout or insert lncRNAs and the influence of lncRNAs on aneurysm rupture would analysed; 2) the influence of lncRNAs on macrophage polarization would analysed; 3) confirming the regulation relationship between lncRNAs and micRNAs, our study would focus on whether the long chain noncoding RNA played a role of aneurysm rupture dependent on miRNA regulation and macrophage polarization as ceRNA mechanism. This topic proposed long chain non-coding RNA in intracranial aneurysm rupture mechanism research, offering a new theoretical basis for the diagnosis and treatment of the disease.

巨噬细胞M1型/M2型极性分化失衡是导致颅内动脉瘤(IA)破裂的重要原因，而miRNAs在巨噬细胞极性分化过程中发挥重要作用。现有研究发现，长链非编码RNA（lncRNAs）可作为竞争性內源RNA(ceRNA)，对miRNAs活性进行有效调控；但此机制是否存在于调控巨噬细胞极性分化过程，进而影响颅内动脉瘤破裂，还有待证实。本课题组前期已通过基因芯片技术筛选出人脑动脉瘤组织和巨噬细胞极性分化过程中差异性表达的lncRNAs与miRNAs，拟在此基础上深入研究：1）将大鼠进行lncRNAs敲除或敲入后，构建IA模型中，检测lncRNAs对动脉瘤破裂的影响；2）检测lncRNAs对巨噬细胞极性分化的影响；3）探讨lncRNAs与miRNAs的靶向调控关系，明确lncRNAs作为ceRNA对动脉瘤破裂的影响，依赖于对巨噬细胞极性分化的调控，并探索其机制。为IA的诊断和治疗提供新的理论基础。

巨噬细胞M1型/M2型极性分化失衡是导致颅内动脉瘤(IA)破裂的重要原因，而miRNAs在巨噬细胞M1/M2极性分化过程中发挥重要作用。现有研究发现，长链非编码RNA（lncRNAs）可作为竞争性內源RNA(ceRNA)，对miRNAs活性进行有效调控；但此机制是否存在于调控巨噬细胞极性分化过程，进而影响颅内动脉瘤破裂，还有待证实。本课题组前期已通过基因芯片技术筛选出人脑动脉瘤组织和巨噬细胞极性分化过程中差异性表达的lncRNAs与miRNAs，将大鼠进行lncRNAs敲除或敲入后，构建IA模型中，检测lncRNAs对动脉瘤破裂以及对巨噬细胞M1/M2极性分化的影响；探讨lncRNAs与miRNAs的靶向调控关系，明确lncRNAs作为ceRNA对动脉瘤破裂的影响，并验证这一过程是否依赖巨噬细胞极性分化。结果显示：（1）lncRNA LOC643401与miR-146a在颅内动脉瘤（IA）患者动脉瘤壁组织，以及巨噬细胞M1/M2极性分化过程中存在差异性表达。（2）在巨噬细胞细胞水平中，通过荧光素酶报告、RNA pull down等发现，lncRNAs LOC643401可作为ceRNA，竞争性结合miR-146a，降低后者活性，解除其对下游靶基因INHBA的抑制作用。（3）对巨噬细胞极性分化，lncRNAs LOC643401可作为ceRNA，竞争性结合miR-146a，促进巨噬细胞由M2向M1型分化。（4）大鼠颅内动脉瘤模型中，lncRNA LOC643401可作为ceRNA，通过抑制miR-146a活性，促进颅内动脉瘤破裂。（5）lncRNA LOC643401、miR-146a、miRNAs下游靶基因INHBA与颅内动脉瘤破裂的具有相关性。降低lncRNA LOC643401含量，可减少IA破裂机率。（6）抑制巨噬细胞M1比例后，过表达lncRNA LOC643401对颅内动脉瘤破裂的促进作用减弱。