眶内针刺对前部缺血性视神经病变基因表达谱的影响及其网络模块化分析

Anterior ischemic optic neuropathy (AION) is an eye disease with high incidence of severe disability and blindness affected by systemic and local factor, which are more and more concerned by ophthalmologist. For treating AION, it was demonstrated by our team that effective rate had been up to 61.1% by using acupuncture treatment. However, it is well known that pathological process of AION is extremely complex, which leads to a bottleneck that trying in vain to uncover the treatment mechanism by single target or pathway. Fortunately, the development of post-genome (functional genome) and biochip technology and modular analysis method of complex biological networks may provide a novel paradigm for mechanism comprehension. Briefly, after treating diseased model, we can get the gene expression data by microarray experiment and screen out the differential expression genes for further confirming the therapeutic targets. These targets here are using for constructing the molecular network. For this network, both of module division and module function annotation are aimed at identifying the functional module of treatment and making the therapeutic mechanism clear. Consequently, it is an opportunity for our team to dissect the multi-target intervention mechanism of intraorbital acupuncture in the treatment of AION through analyzing the change of gene expression profiles and network modules. We suggest that combining multifactorial pathology of AION with multi-target intervention analysis of acupuncture may help to promote the application of acupuncture in clinical ophthalmology.

前部缺血性视神经病变（AION）作为受全身、局部因素均影响较大的临床高发、严重致残、致盲性眼病受到医者的关注。本课题组通过前期临床研究证实针刺治疗AION有效率达61.1%。然而，众所周知，AION发生的病理过程是极其复杂的，单靶点或单通路的研究揭示系统的治疗机制存在瓶颈。后基因组（功能基因组）、基因芯片技术的发展以及复杂生物网络的模块化分析思路为我们的研究提供了曙光。简言之，就是对疾病模型干预后，利用基因芯片技术获得基因表达谱数据，筛选出差异表达基因明确干预靶点，运用计算机构建疾病干预的靶点相关网络，并对该网络进行模块划分及功能注释，从而，明确该治疗方法的功能模块及治疗机制。因此，本课题旨在通过对眶内针刺干预AION基因表达谱的影响及网络模块化分析，明确该疗法对AION多靶点干预机制。多因素致病特点的AION个体与针刺干预多靶点分析的结合，将加速推动针刺疗法在眼科临床中的应用。

基于前期对针刺治疗AION临床观察的有效性，结合该病复杂发病机制，我们对AION大鼠模型采用眶内针刺治疗，通过对视网膜神经节细胞（RGCs）的组织学观察，利用转录组测序及系统生物学分析技术，探索针刺干预机制。正常组、模型组、针灸组大鼠视网膜单象限及组合象限区域性RGCs计数比较具有显著性差异，提示眶内针刺组能够有效减少NAION大鼠RGCs的丢失（P＜0.05），但计数仍显著低于正常组（P＜0.05）。对3组大鼠视网膜进行转录组测序，模型组（较正常组）、针灸组（较模型组）分别获得了351、462个差异表达基因，两组重叠的主要差异表达基因包括MYL9、MYH11、THBD、DCN、RPS6、FOS、KRT5、NDUFS6、EEF1A1等，且针灸组显示出对上述基因的逆转表达。功能涉及抗血管收缩、抑制自噬、减少缺血再灌注损伤、增加视神经细胞机械应力、改善线粒体功能及提高突触后抑制。我们基于差异表达基因构建NAION模型——针灸分子间相互作用网络，并对该进行模块划分及筛选主要模块，对主要基因、信号通路及主要模块进行功能分析，机制可能涉及增加视神经细胞机械应力、RGC的存活与稳态、轴突生长、抗凝与抗血管收缩、抑制自噬、减少缺血再灌注损伤、改善线粒体功能及提高突触后抑制。