Using RNA-Seq to Explore the Repair Mechanism of the Three Methods and Three-Acupoint Technique on DRGs in Sciatic Nerve Injured Rats
Using RNA-Seq to Explore the Repair Mechanism of the Three Methods and Three-Acupoint Technique on DRGs in Sciatic Nerve Injured Rats
复制标题
利用RNA-Seq探讨三法三穴技术对坐骨神经损伤大鼠背根神经节的修复机制
DOI：
10.1155/2020/7531409
复制
发表时间：
2020-06-08
影响因子：
2.9
通讯作者：
Steven, Wong
中科院分区：
文献类型：
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作者：
Lv, Tao-tao;Mo, Yan-jun;Steven, Wong
Objective.研究三法三穴法对SNI模型大鼠DRG基因表达的影响，探讨三法三穴法促进周围神经损伤修复的分子机制。将27只雄性SD大鼠随机分为3组：假手术组（Sham）、SNI组（SNI组）和推拿组（推拿组）。推拿组采用推拿手法模拟器模拟穴位按摩，控制质量和数量。于造模后7 d开始，分别于患侧阴门、承山、阳陵泉穴定量给予点压、拨揉、揉法，每日1次。干预1次/d，共10 d，休息1 d，再干预10 d，共干预20次。采用行为学分析评价三种方法和三点法对损伤大鼠恢复的影响。同时对3组大鼠背根节差异表达基因进行RNA测序（RNA-Seq）分析。GO和KEGG富集进行了分析，并使用实时PCR验证。RNA-Seq结合数据库信息显示，推拿组与SNI组相比，DRG差异表达基因数量最多，共226个。GO功能在细胞过程、离子结合、蛋白质结合、神经元、对压力的响应、对金属离子的响应、神经元投射和其他生物过程的正调节中富集。GO功能也在KEGG数据库中的Wnt、IL-17和MAPK信号通路中富集。PCR结果与RNA测序结果一致，提示转录组测序结果可靠。三种方法和三穴法均能通过改变DRG基因序列促进SNI模型大鼠的恢复。
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.