miR-H6对角膜HSV-1感染后炎症反应的效应和机制研究

Herpes simplex virus type 1(HSV-1) keratitis is one of the mainly cause of the blindness worldwide. MicroRNAs are small, 21-23 nucleotide non-coding RNAs that play an important role in the post-transcriptional regulation of gene expression in a wide range of organisms from unicellular eukaryotes to multicellular eukaryotes by a variety of mechanisms. Up to mow, 17 HSV-1 miRNAs have been found and miR-H6 showed high level in HSV-1 lytic infection. Our previous studies shows that miR-H6 can inhibit HSV-1 replication in human corneal epithelial cells and decrease the interleukin 6 released into the medium. In addition,miR-H6 reduced corneal ulcer in HSV-1 infected mice, while 10 target genes including interleukin 8 receptor in host cells were identified by CYTOSCAPE software. Based on the literature and our previous studies, we are going to further explore the effect of miR-H6 on corneal inflammation after HSV-1 infection and possible mechanism in the HSV-1 infected cells and mouse: (1) analysis the effect of miR-H6 on HSV-1 replication in mice cornea and tissue pathology; (2) research the the effect of HSV-1 miR-H6 on host cellular gene by Digital Gene Expression Profiling gene technology and verified its target genes; (3)observe the effect of HSV-1 miR-H6 on the host inflammatory response,especially IL-6 and IL-8, in mice of viral keratitis.The results will be helpful to elucidate the pathological mechanism of HSV-1 keratitis, at the same time, provide the theoretical and experimental evidence of virus miRNA participation in host gene regulation,and eventually used in the treatment of viral infectious diseases.

Ⅰ型疱疹病毒(HSV-1)性角膜炎是常见致盲眼病。病毒感染中，MicroRNA(miRNA,miR)参与基因表达调控等多种生物过程。我们前期实验发现HSV-1的miR-H6可以抑制细胞中HSV-1复制并减少IL-6的释放，减小病毒感染小鼠的角膜溃疡灶，同时软件预测发现miR-H6在宿主细胞存在10个靶基因（IL-8受体等）。本课题拟在HSV-1感染的细胞及小鼠角膜炎模型中进一步研究miR-H6对角膜HSV-1感染后炎症反应的效应和机制，包括：①分析miR-H6对HSV-1感染角膜中病毒复制及组织病理变化；②利用芯片技术研究miR-H6对宿主基因表达的影响，并验证其靶基因；③针对性观察miR-H6对小鼠病毒性角膜炎中IL-6，IL-8等炎症因子相关的免疫炎症的影响。结果将有助于阐明HSV-1角膜炎的病理机制，同时也对病毒miRNA参与调控宿主基因提供理论及实验证据，最终用于治疗病毒感染性疾。

Ⅰ型疱疹病毒(HSV-1)性角膜炎是常见致盲眼病。我们前期实验发现针对HSV-1 ICP4的小干扰RNA可以抑制细胞中HSV-1复制并减少IL-6的释放，减小病毒感染小鼠的角膜溃疡灶。本课题在前期工作基础上发现1）革兰氏染色阴性菌的脂多糖（LPS）可以促进上皮细胞感染HSV-1，而革兰氏染色阳性菌成份（金黄色葡萄球菌裂解液）可以抑制上皮细胞感染HSV-1，这为探索眼表微环境变化对HSV1感染的影响奠定理论基础。2）结合临床工作，由于微生物入侵眼内组织生长繁殖引起的炎症反应统称为感染性眼内炎。最终可能累计眼的各种结构，严重威胁患者的视力。我们分析了我院2010年1月起共5年的330眼内炎患者的致病微生物及耐药性，发现细菌对左氧氟沙星最为敏感，为临床诊治眼内炎提供一定的依据。3）我们总结了我院十年内培养阳性的内源性真菌性眼内炎的临床特点，发现初始视力低于指数的患者预后较差，霉菌比酵母菌的预后差。4）糖尿病视网膜病变也是主要的致盲眼病，我们通过定型研究的方法发现影响患者未及时就诊的因素包括缺乏糖尿病视网膜病变的相关知识，对使用胰岛素的顾虑，家庭和社会关系的相互影响以及疾病治疗的经济负担。我们的研究结果为防治糖尿病视网膜病变提供一定的指导价值。5）危急值是指某项或某类检验异常结果，而当这种检验异常结果出现时，表明患者可能正处于有生命危险的边缘状态。我们总结并分析了我院2012到2014年的血液检查的危急值，发现眼科患者危急值的发生率为5.7‰，并与患者年龄和疾病类型有关。. 经本项目资助，目前已发表4篇SCI 论文，另1篇文章已投稿待发表；参加国内外会议3 人次。