电针干预对单眼形觉剥夺弱视大鼠视皮层基因表达的影响

Effect of electroacupuncture intervention on gene expression in visual cortex of monocular form deprivation amblyopia rats

目的运用基因芯片技术对形觉剥夺及电针干预后的弱视大鼠视皮层中差异表达基因进行生物学分析与关键基因的筛选,为弱视发生及电针治疗提供基因水平理论依据。方法将SD大鼠随机分为正常组、弱视组和电针组,每组5只。弱视组和电针组大鼠采用单眼缝合方法造成右眼剥夺性弱视,术后30~60 d对电针组大鼠进行电针干预,每周至少干预6 d。电针干预结束后,处死各组大鼠并取左侧视皮层,使用基因芯片技术检测视皮层中基因表达情况。筛选各组中Fold Change≥2.0且P<0.05的差异基因进行统计,应用基因本体论(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析差异基因的功能及其参与的生物学通路。结果与正常组比较,弱视组有135个基因下调,28个基因上调;与弱视组比较,电针组有52个基因上调,25个基因下调。GO富集分析发现弱视组和电针组产生的差异基因主要功能集中于蛋白翻译、蛋白转运、离子跨膜转运、细胞内外小分子结合、组织器官发育等。KEGG富集分析发现弱视组和电针组产生的差异基因共同参与ERBB信号通路、光信号传导、TLR信号通路、神经胶质细胞瘤等。PPI网络显示丝裂原活化蛋白激酶1(MAPK1)和热休克蛋白A1A(HSPA1A)基因位于网络核心,二者的异常表达可能与弱视发生及电针干预密切相关,在神经突触可塑性过程中起重要作用。结论形觉剥夺弱视大鼠的视皮层在视觉发育过程中存在大量基因的异常表达,电针刺激可以拮抗部分基因的差异表达;由MAPK1和HSPA1A基因调控的代谢通路改变可能是弱视发病及电针治疗弱视的重要分子生物学机制。

Objective Using gene chip technology to biologically analyze differentially expressed genes in the visual cortex of amblyopia rats after form deprivation and electroacupuncture intervention,and screen for key genes,providing a genetic level theoretical basis for the occurrence of amblyopia and electroacupuncture treatment.Methods SD rats were randomly divided into normal group,amblyopia group,and electroacupuncture group,with 5 rats in each group.The rats in the amblyopia group and the electroacupuncture group were subjected to unilateral suture to cause right eye deprivation amblyopia.Within 30-60 days after surgery,the rats in the electroacupuncture group received electroacupuncture intervention for at least 6 days per week.All rats were killed at the same time after modeling and left visual cortex was taken.Gene chip technology was used to detect gene expression in visual cortex of each sample.In each group,samples were selected for statistics of differential genes with Fold Change≥2.0 and P<0.05,and the functions of differential genes and the biological pathways involved in them were analyzed by Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Results Compared with the control group,135 genes were down regulated and 28 genes were up regulated in the amblyopia model group;Compared with amblyopia model group,52 genes were up-regulated and 25 genes were down regulated in electroacupuncture treatment group.GO enrichment analysis found that the main functions of differential genes generated after amblyopia modeling and electroacupuncture treatment were protein translation,protein transport,ion transmembrane transport,small molecule binding inside and outside cells,tissue and organ development,etc.KEGG enrichment analysis found that the differential genes generated after amblyopia modeling and electroacupuncture treatment were involved in ERBB signal pathway,light signal transduction,TLR signal pathway,glioma,etc.PPI network showed that MAPK1 and HSPA1A genes were located at the core of the network,and their abnormal expression might be closely related to the occurrence of amblyopia and the improvement process of electroacupuncture,which played an important role in the process of neural synaptic plasticity.Conclusion There is a large amount of abnormal gene expression in the visual cortex of form deprivation amblyopic rats during visual development,and electroacupuncture stimulation can antagonize the differential expression of some genes.The changes in metabolic pathways regulated by MAPK1 and HSPA1A genes may be an important molecular biological mechanism for the pathogenesis of amblyopia and the treatment of amblyopia with electroacupuncture.