Optic neuritis (ON) is one of the most frequent symptoms of multiple sclerosis (MS) that results in progressive loss of axons and neurons. In clinical trials of Traditional Chinese Medicine, needling at the GB20 acupoint has been widely used for the treatment of ocular diseases, including ON. However, the molecular mechanisms of needling at this site are still unclear. In this study, we generated an experimental autoimmune encephalomyelitis (EAE) mouse model and investigated the effects of needling treatment at the GB20 acupoint on retina with EAE-associated ON. RNA sequencing of the retinal transcriptome revealed that, of the 234 differentially expressed genes induced by ON, 100 genes were upregulated, and 134 genes were downregulated by ON, while needling at the GB20 acupoint specifically reversed the expression of 21 genes compared with control treatment at GV16 acupoint. Among the reversed genes, Nr4a3, Sncg, Uchl1, and Tppp3 were involved in axon development and regeneration and were downregulated by ON, indicating the beneficial effect of needling at GB20. Further gene ontology (GO) enrichment analysis revealed that needling at GB20 affected the molecular process of Circadian rhythm in mouse retina with ON. Our study first reported that needling treatment after ON at the GB20 acupoint regulated gene expression of the retina and reversed the expression of downregulated axon development-related genes. This study also demonstrated that GV16 was a perfect control treatment site for GB20 in animal research. Our study provided a scientific basis for needling treatments at GB20 for ocular diseases.

视神经炎（ON）是多发性硬化症（MS）最常见的症状之一，可导致轴突和神经元进行性丧失。在中医药的临床试验中，针刺GB20穴位已被广泛用于治疗包括ON在内的眼科疾病。但是，在该部位进行针刺的分子机制仍不清楚。在这项研究中，我们生成了一个实验性自身免疫性脑脊髓炎（EAE）小鼠模型，并研究了与EAE相关的ON对GB20穴位进行针刺治疗对视网膜的影响。视网膜转录组的RNA测序显示，在ON诱导的234个差异表达基因中，有100个基因被上调，而134个基因被ON下调，与在GV16穴位上的对照治疗相比，在GB20穴位上进行针刺可特异性逆转21个基因的表达。在反向基因中，Nr4a3，Sncg，Uchl1和Tppp3参与轴突的发育和再生，并被ON下调，表明在GB20处针刺的有益作用。进一步的基因本体论（GO）富集分析表明，GB20处的针刺会影响ON小鼠视网膜中昼夜节律的分子过程。我们的研究首次报道了在GB20穴位ON进行针刺治疗后，调节了视网膜的基因表达，并逆转了下调的轴突发育相关基因的表达。这项研究还表明，GV16是动物研究中GB20的理想对照治疗位点。