汉族人群血管内皮生长因子基因多态性与脑微出血相关性及其分子机制研究

Cerebral microbleeds (CMBs) are important risk factors for stroke hemorrhagic transformation, prognosis and recurrence, resulting in cognitive impairment and age-related loss of the ability. There is no effective means of prevention and treatment for them up till now. Symptomatic hemorrhage transformation is the main bottleneck of breaking through therapeutic time window of thrombolytic therapy for cerebral infarction.Therefore, screening for the risk marker of predicting CMBs and exploreing its pathogenesis is one of the key issues for solving the aforementioned medical problems. Studies have shown that VEGF is involved in the pathological process of CMBs through several signaling pathways. Our previous study has showed that the serum VEGF levels significantly increase in the patients with cerebral infarction complicated by CMBs, and individuals with high serum levels of VEGF are more susceptible to CMBs. It is known that VEGF gene expression and transcription activity is regulated by VEGF gene polymorphisms. The aim of this study is to explore the correlation between VEGF gene polymorphisms and acute cerebral infarction complicated by CMBs in Han Chinese population by use of gene sequencing, gene targeting mutations, gel shift assay and chromatin immunoprecipitation methods and so on. Based on the APOE gene genotyping, interaction effects between susceptibility genes would be analyzed. The functional verification for the promoter region of SNPs (significantly correlated to CMBs) would be performed as well as its regulatory mechanism. This project would contribute to clarify the molecular mechanism of acute cerebral infarction complicated by CMBs, and provide a point of penetration for the prevention and treatment of CMBs and breakthing rough therapeutic time window of thrombolytic therapy for cerebral infarction..

脑微出血(CMBs)是影响脑卒中出血转化、预后及复发、造成认知障碍及年龄相关能力丧失的重要危险因素，尚无有效的防治手段。症状性出血转化是突破脑梗死溶栓时间窗的主要瓶颈。因此，筛查CMBs发生风险相关标志物并探讨其发生机制是解决上述医学难题的关键问题之一。VEGF通过多条信号通路参与CMBs的病理过程。前期研究发现脑梗死合并CMBs的患者血清VEGF水平升高，高血清VEGF水平个体更易发生CMBs。已知VEGF基因多态性影响其基因表达和转录活性。本项目拟采用基因测序、基因定点突变、凝胶阻滞电泳及染色质免疫共沉淀等技术，探讨VEGF基因多态性与汉族人急性脑梗死合并CMBs的相关性，结合APOE基因分型，进行易感基因之间的交互效应分析，并对启动子区SNPs（与CMBs关联）进行功能验证及调控机制研究。本项目将阐明急性脑梗死发生CMBs的分子机制，为CMBs的防治及突破脑梗死溶栓时间窗提供切入点。

脑小血管病研究已成为当前的热点研究领域。本项目按照原先的研究计划对VEGF 基因多态性与血清VEGF表达水平的关系进行研究，并未发现与VEGF表达存在明确关联的VEGF基因多态位点。探讨了VEGF基因多态性及血清VEGF水平与脑微出血相关性。在已取得良好的分子流行病学研究基础上，紧紧围绕着国家重点支持的脑小血管病研究领域的热点问题，进行了相应的拓展研究。旨在探寻脑小血管病发生风险、病变结构及功能病损的影像学评估关键技术。项目组开展了不同影像学技术评估脑微出血对脑梗死患者出血转化的预测价值研究。进行了脑微出血生物学标记物预警诊断筛查研究工作，发现了血清VEGF与尿液AD7c-NTP可能是脑微出血的风险标志物。对脑小血管病的发病机制、发病的风险因素及预测进展性运动功能缺损的生物学标记物方面进行了更多的探讨，初步揭示了脑梗死进展的影像学标志物。探讨了脑小梗死影像学模式与血压变异性的相关性，发现了长时血压变异性与、短时血压变异性与脑小梗死影像学模式特征的对应关系，为脑小血管病的风险因素的控制及血压管理策略的完善提供有用的研究证据。在拓展研究中，对缺血性脑血管病的发病机制理论及预防控制理念进行总结分析，提出了创新性的理论假说和观点。本项目研究成果包括：发表或已录用23篇论文（包括会议论文），举办一次市级学术会议。获得市级科研奖励3项，项目组成员获得地市级青年科技奖1人次，获得地市级医疗卫生优秀骨干人才1人次，入选地市级122工程人才培养计划1人次，入选省级高层次人才培养计划1人次。资助5名硕士研究生和1名博士研究生进行学位课题研究。