白细胞介素-6对人脑动静脉畸形细胞连接的影响及机制研究

IL-6 and MMP-9 expression of ruptured cerebral arteriovenous malformation in the tissues was significantly higher than the expression unruptured group; And MMP-9 was reported degrading membrane protein parts of cell junction of neurons. Thus we assume that IL-6 May through some MARK pathways or oxidative stress reaction makes cell membrane protein degradation of the connection in cells, microfilaments polymerization increase and the tube protein depolymerization increase.all these lead to cell connection damaged,thus cause blood vessels to instability. To prove this assumption we establish the primary cell line of the brain arteriovenous malformation endothelial cell model, with IL-6 cocultured, and on the basis of the original crack with various silk amp-activated protein kinase signal pathway (ERK, P38 and JNK) inhibitors, testing the reaction of after ERK, P38, JNK, MMP-9, cell connection protein (VE-cad herin, claudin, ZO-1) expression and cell actin and microtubules polymerization reaction, and gene chip method for determining arteriovenous malformation cell connection and cytoskeleton gene expression. Testing the plasma interleukin -6 level of cerebral arteriovenous malformation patients before and after surgery . Aims to show that basis on the influence of IL-6 and cytoskeleton gene downexpression,the stability fo endothelial cell wall of cerebal arteriovenous malformation , provide new targets for clinical treatment.

IL-6和MMP-9在破裂脑动静脉畸形（cAVM）组织中的表达明显高于未破裂组且MMP-9能降解神经元细胞连接膜外蛋白部分；电镜示畸形组织细胞连接减少；肌动蛋白相关基因WASL在遗传性出血性毛细血管扩张症患者中下调，由此我们假设在cAVM细胞连接相关基因表达下调基础上IL-6可能通过某种MARK通路或氧化应激反应使细胞连接受损。为证实假设，我们在前期原代培养的cAVM内皮细胞模型基础上，用IL-6共培养畸形内皮细胞，加入各种丝裂原活化蛋白激酶信号通路（ERK,P38和JNK）的抑制剂，检测各组反应后的ERK,P38,JNK,MMP-9,细胞连接蛋白（VE-cadherin、cingulin,ZO）的表达以及细胞肌动蛋白和微管聚合反应，并用基因芯片测定AVM细胞连接和细胞骨架相关基因表达。检测患者手术前后时期血液中IL-6水平的变化。旨在为临床工作提供新的药物靶点及为畸形出血提供新的预警指标

收集天坛医院手术切除的脑动静脉畸形患者的畸形组织以及手术同侧的颞浅动脉组织各4份，其中男3例，女1例。标本组织处理后进行如下实验内容 1 长链非编码RNA数据分析1）长链非编码RNA的提取和标准化2）长链非编码RNA的标本质量控制3）长链非编码RNA的差异性分析4）热图以及层聚分析 2 mRNA数据分析 1）mRNA的提取和标准化2）mRNA的标本质量控制3）mRNA的差异性分析4）热图以及层聚分析 5）通路分析 6）基因生物学分析 3 长链非编码RNA 1）增强子分析 2）长链非编码RNA全图分析 3）hox聚类图分析 4）长链非编码RNA邻近编码基因分析 5）增强子长链非编码RNA邻近编码基因分析。从初步得到的结果提示：每组配对的通路分析中，与nfkb mapk有关的hsa04060，hsa04064以及hsa04010有统计学差异。目前大量数据正处理中，随后还有根据数据的分析结果继续进行相关的验证实验工作