以RNA-Seq技术探究推拿三法三穴对minorCCI模型大鼠的镇痛作用及启动机制

Discussion on the analgesic effect and starting mechanism of three-manipulation and three-acupoint on minor CCI model rats via RNA sequencing

目的:探究推拿三法三穴对轻型慢性压迫性神经损伤(minorCCI)模型大鼠的即刻镇痛作用及启动机制。方法:将54只SD大鼠随机分为:假手术1组、2组、3组,模型1组、2组、3组,推拿1次组、2次组、3次组,每组6只。模型组和推拿组建立minorCCI模型。推拿各亚组在造模后7d进行三法三穴干预1次、2次、3次,最后一次推拿干预结束即刻进行机械缩足反射阈值(MWT)和热缩足反射潜伏期(TWL)测试,测试结束取腰膨大节段脊髓背角(SDH)进行转录组测序(RNA-Seq)研究。结果:推拿1次、2次、3次后均可提高MWT和TWL测试结果(P<0.05)。RNA-Seq结果显示,符合时间序列表达显著变化趋势的差异表达基因(DEGs)共有118个,富集到神经营养素信号通路、细胞凋亡、PI3K-Akt信号通路、Ras信号通路、JAK-STAT信号通路等111条通路上。结论:推拿1次具有即刻镇痛作用,基因层面的推拿镇痛启动可能在推拿1次后,推拿镇痛启动级联反应机制可能在推拿2次后。

Objective:To investigate the immediate analgesic effect,analgesic initiation,and mechanism of the threemanipulation and three-acupoint'of Tuina on minor chronic constriction injury(minor CCI)model rats.Methods:Fifty-four SD rats were randomly divided into sham groups 1,2,and 3,model groups 1,2,and 3,and Tuina groups 1,2,and 3,with 6 rats in each group.The model and Tuina groups were established with minor CCI models.The subgroups of Tuina were subjected to‘three-manipulation and three-acupoint'for 1,2,and 3 times after 7 days of modeling,and the mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)tests were performed immediately after the last Tuina intervention,and the spinal dorsal horn(SDH)of the lumbar dilated segment was taken at the end of the test for RNA sequencing(RNA-Seq)study.Results:MWT and TWL results were improved after 1,2,and 3 times of Tuina(P<0.05).Analysis of RNA-Seq results showed a total of 118 differentially expressed genes(DEGs)consistent with a significant trend in time-series expression,enriched for 111 pathways including neurotrophin signaling pathway,apoptosis,and PI3K-Akt signaling pathway,Ras signaling pathway,and JAK-STAT signaling pathway.Conclusion:1-time Tuina has an immediate analgesic effect,the initiation of Tuina analgesia at the genetic level may be after 1-time Tuina,and the mechanism of Tuina analgesia initiation cascade response may be after 2-time Tuina.