Effect of acupuncture treatments on retinal transcriptome after optic nerve injury

Purpose: We used RNA sequencing to investigate the effect of acupuncture treatment on retinal transcritome after optic nerve injury Methods: Retinal mRNA profiles of 5-week-old wild-type (WT), optic nerve crush injury (ONC), ONC injury with acupuncture treatment at acupoint GB20 (ONC-F) and ONC injury with acupuncture treatment at acupoint BL1 (ONC-J) mice were generated by deep sequencing, in triplicate, using Illumina Hiseq platform. The sequence reads that passed quality filters were analyzed with DESeq2 R package to identify differently expressed genes with padj<0.05. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: RNA-seq revealed that 436 genes including 31 transcription factors (TFs) were changed after injury, which include many well-known neural degeneration related TFs such as Jun, Ddit3, Atf3 and Atf4. Interestingly, acupuncture treatment at acupoint GB20 (Fengchi) or BL1(Jingming) after ONC injury both resulted in change of retinal gene expression, compared with ONC injury without treatment; furthermore, acupuncture treatment also reversed several gene expression status induced by axon injury. Conclusions: Our research first reported that acupuncture treatment after injury can regulated retinal transcriptome and reversed the gene expression level induced after injury, which will provide novel therapeutic targets for treatment of retinopathy diseases. Retinal mRNA profiles of 5-week-old mice were generated by deep sequencing, in triplicate, using Illumina Hiseq.

研究目的：本研究采用RNA测序技术，探究视神经损伤后针刺治疗对小鼠视网膜转录组的影响。
研究方法：针对5周龄野生型（wild-type，WT）小鼠、视神经钳夹损伤（optic nerve crush injury，ONC）模型小鼠、ONC模型联合GB20穴位针刺干预小鼠（ONC-F）、ONC模型联合BL1穴位针刺干预小鼠（ONC-J），采用Illumina Hiseq平台对其视网膜mRNA表达谱进行深度测序，每组设置3次生物学重复。对通过质量过滤的测序reads，使用DESeq2 R包进行数据分析，筛选校正后P值（padj）<0.05的差异表达基因。采用TaqMan探针法与SYBR Green染料法开展qRT-PCR验证实验。
研究结果：RNA测序结果显示，视神经损伤后共有436个基因发生表达改变，其中包含31个转录因子（transcription factors，TFs），包含Jun、Ddit3、Atf3、Atf4等多种经典神经退行性相关转录因子。值得注意的是，与未接受治疗的ONC模型小鼠相比，ONC损伤后分别于GB20（风池）或BL1（睛明）穴位进行针刺干预，均可改变小鼠视网膜的基因表达模式；此外，针刺治疗还可逆转轴突损伤诱导的部分基因表达状态。
研究结论：本研究首次报道，损伤后针刺治疗可调控视网膜转录组，并逆转损伤诱导的基因表达水平变化，该研究成果将为视网膜疾病的临床治疗提供全新的潜在靶点。本研究通过Illumina Hiseq平台对5周龄小鼠的视网膜mRNA表达谱进行了深度测序，每组设置3次生物学重复。