Objective. To study the effects of the three methods and three-acupoint technique on DRG gene expression in SNI model rats and to elucidate the molecular mechanism of the three methods and three-acupoint technique on promoting recovery in peripheral nerve injury. Methods. 27 male SD rats were randomly divided into three groups: a Sham group, the SNI group, and the Tuina group. The Tuina group was treated with a tuina manipulation simulator to simulate massage on points, controlling for both quality and quantity. Point-pressing, plucking, and kneading methods were administered quantitatively at Yinmen (BL37), Chengshan (BL57), and Yanglingquan (GB34) points on the affected side once a day, beginning 7 days after modeling. Intervention was applied once a day for 10 days, then 1 day of rest, followed by 10 more days of intervention, totally equaling 20 times of intervention. The effect of the three methods and three-point technique on the recovery of injured rats was evaluated using behavior analysis. RNA sequencing (RNA-Seq) analysis of differentially expressed genes in DRGs of the three groups of rats was also performed. GO and KEGG enrichment was analyzed and verified using real-time PCR. Results. RNA-Seq combined with database information showed that the number of differentially expressed genes in DRG was the largest in the Tuina group compared with the SNI group, totaling 226. GO function is enriched in the positive regulation of cell processes, ion binding, protein binding, neuron, response to pressure, response to metal ions, neuron projection, and other biological processes. GO function is also enriched in the Wnt, IL-17, and MAPK signaling pathways in the KEGG database. PCR results were consistent with those of RNA sequencing, suggesting that the results of transcriptome sequencing were reliable. Conclusion. The three methods and three-acupoint technique can promote the recovery of SNI model rats by altering the gene sequence in DRGs.

中文翻译：

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利用RNA-Seq探索三种方法和三穴技术对坐骨神经损伤大鼠DRGs的修复机制。

目标。研究三种方法和三穴技术对SNI模型大鼠DRG基因表达的影响，阐明三种方法和三穴技术促进周围神经损伤恢复的分子机制。方法。将27只雄性SD大鼠随机分为三组：假手术组，SNI组和推拿组。推拿小组接受了推拿操纵模拟器的治疗，以模拟按摩点，同时控制质量和数量。从建模后的7天开始，每天对患侧的银门（BL37），城山（BL57）和杨凌泉（GB34）点定量施压，拔毛和揉捏方法。每天进行一次干预，持续10天，然后休息1天，然后再干预10天，总计等于干预20次。使用行为分析评估了三种方法和三点技术对受伤大鼠恢复的影响。还对三组大鼠的DRG中差异表达的基因进行了RNA测序（RNA-Seq）分析。使用实时PCR分析和验证GO和KEGG富集。结果。RNA-Seq结合数据库信息显示，与SNI组相比，Tuina组中DRG中差异表达基因的数量最多，总计226个。GO功能丰富于细胞过程，离子结合，蛋白质结合的正调控中，神经元，对压力的反应，对金属离子的反应，神经元投射和其他生物过程。GO功能还丰富了KEGG数据库中的Wnt，IL-17和MAPK信号通路。PCR结果与RNA测序结果一致，表明转录组测序结果可靠。结论。三种方法和三穴技术可以通过改变DRGs中的基因序列来促进SNI模型大鼠的恢复。