脑出血患者外周血及出血灶周组织特异性microRNA表达谱及其脑水肿机制的研究

Spontaneous intracerebral hemorrhage is one of significant causes of morbidity and mortality throughout the world. Cerebral edema after hemorrhage is the main factor affecting the prognosis. However, in clinical practice, we are still lack of non-invasive and effective treatment of cerebral edema to date. In this study, we focus on the microRNAs expressed both in the brain and in the peripheral blood of patients with cerebral edema utilizing oligonucleotide microarray. The selected microRNAs should be validated by quantitative real-time polymerase chain reaction. In addition, the relationship of the microRNAs between the brain tissue and the peripheral blood will be analysed. On this basis, using bioinformatic method, the prediction of the microRNAs target genes and selection of the microRNAs whose target genes are AQP-1, AQP-4, VCAM-1 or MMP-9 will be performed. At last, at the cellular level, the relationship between selected microRNAs and their target genes will be testified by luciferase reporter assay. The purpose of our study is to uncover the microRNA as the special biological marker in the blood of patients and the role of microRNAs in the molecular pathogenesis of cerebral edema in order to explore a new direction in clinical monitrition and treatment.

近年来脑出血患者死亡率、致残率始终位于高位，而出血后脑水肿则是影响预后的主要因素。针对脑水肿的治疗目前尚缺乏无创而有效的手段。本课题对脑出血水肿期患者应用芯片杂交的方法检测其外周血及出血灶周组织特异性microRNA表达谱，以发现特异性表达的microRNA，并应用实时荧光定量PCR对其进行进一步验证；分析外周血和灶周组织microRNA表达谱之间是否具有同步相关性。在此基础上，通过生物信息学软件预测特异性表达的microRNA可能的靶基因；挑选出预测靶基因为AQP-1、AQP-4、VCAM-1和MMP-9的microRNA，最后在细胞水平通过构建荧光素酶报告系统证实二者之间的关系，以期望发现外周血中可预测脑水肿的特异性microRNA类生物标记物，阐明脑出血后水肿可能的分子机制，从而为脑出血后灶周水肿的监测和治疗提供理论依据。

近年来脑出血患者死亡率、致残率始终位于高位，而出血后脑水肿则是影响预后的主要因素，脑水肿目前尚缺乏有效的预测及治疗的手段。本课题主要研究脑出血患者外周血microRNA的表达情况及microRNA与脑出血后脑水肿的机制。我们通过高通量microRNA芯片对脑出血患者外周血中microRNA表达情况进行研究，发现了脑出血患者外周血中存在特异的microRNA表达谱，包含65个具有差异表达的microRNA，其中64个microRNA下调，1个microRNA上调；进一步扩大样本量进行qRT-PCR实验，证实特异性microRNA表达与芯片结果一致；统计分析提示miR-126的表达水平与脑出血后脑水肿呈显著负相关，miR-21-5p等4种microRNA表达水平与血肿体积呈负相关，并总结出microRNA与脑出血时间的相关性；通过生物信息学分析，发现特异性microRNA对脑出血的作用靶点广泛，参与氧化应激、凋亡等脑水肿的病理生理过程。我们相信miR-126可能成为脑出血后脑水肿的预测因子，也有希望成为脑水肿的干预靶点；而miR-21-5p等microRNA可能成为监测及控制血肿体积的手段。脑出血后脑水肿可能与上述特异性microRNA有关，其具体机制有待进一步细胞水平的实验来验证。阐明这一机制将为脑出血后脑水肿的监测和治疗开辟新的方向。