脊柱推拿对“椎骨错缝”大鼠DRG神经元PKCε/TRPV1通路的影响  

Effects of spinal Tuina on PKCε/TRPV1 pathway of DRG neurons in rats with‘mild malposition of vertebra and joint’

作　　者：吕智桢[1,2,3] 程艳彬[4,5] 姚重界 孔令军[5,6] 朱清广[4,5] 吴志伟[4,5] 周鑫 房敏 LYU Zhizhen;CHENG Yanbin;YAO Chongjie;KONG Lingjun;ZHU Qingguang;WU Zhiwei;ZHOU Xin;FANG Min(The 3rd Affiliated Hospital,Zhejiang Chinese Medical University,Hangzhou 310005,China;The 3rd Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou 310005,China;Research Institute of Tuina(Spinal Disease),Zhejiang Chinese Medical University,Hangzhou 310053,China;Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Research Institute of Tuina,Shanghai Academy of Traditional Chinese Medicine,Shanghai 200437,China;Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]浙江中医药大学附属第三医院,杭州310005 [2]浙江中医药大学第三临床医学院,杭州310005 [3]浙江中医药大学推拿脊柱病研究所,杭州310053 [4]上海中医药大学附属岳阳中西医结合医院,上海200437 [5]上海市中医药研究院推拿研究所,上海200437 [6]上海中医药大学附属曙光医院,上海201203

出　　处：《中华中医药杂志》2024年第2期892-896,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金重点项目(No.82030121);中华中医药学会“百千万”人才工程岐黄学者项目(2018);浙江中医药大学校级科研人才专项(No.2021RCZXZK03);浙江省自然科学基金项目(No.LQ23H270010)。

摘　　要：目的:观察腰椎“椎骨错缝”模型大鼠痛行为和背根神经节(DRG)神经元中蛋白激酶Cε(PKCε)、瞬时受体电位香草酸亚型1(TRPV1)和磷酸化TRPV1(p-TRPV1)的影响,探讨脊柱推拿的镇痛效应和机制。方法:将32只大鼠随机分为空白组、假手术组、模型组、手法治疗组,每组8只,通过外部连接固定方法制备“椎骨错缝”模型。手法治疗组大鼠在造模后第15天开始对两侧L3~S1夹脊穴进行5 N、2 Hz、10 min/次、连续14 d的模拟手法治疗。检测各组大鼠不同时间节点机械痛和热痛的改变,取DRG,用免疫荧光技术和Western Blot技术检测PKCε、TRPV1和p-TRPV1表达情况及蛋白含量。结果:造模后,模型组大鼠PWT、PWL均较空白组和假手术组显著下降(P<0.01),干预后第10、14天,手法治疗组大鼠PWT、PWL较模型组显著升高(P<0.01)。免疫荧光显示,模型组与手法治疗组大鼠DRG中PKCε、TRPV1受体表达较治疗前均明显,手法治疗组较模型组PKCε、TRPV1受体表达具有明显抑制趋势。模型组和手法治疗组DRG中PK Cε、TRPV1、p-TRPV1蛋白含量显著高于空白组和假手术组(P<0.01),手法治疗组DRG中PKCε、TRPV1、p-TRPV1蛋白含量显著低于模型组(P<0.05,P<0.01)。结论:脊柱推拿可以改善“椎骨错缝”腰痛大鼠疼痛,其机制可能是抑制PKCε/TRPV1信号通路开放并抑制TRPV1磷酸化而获效。Objective:To observe the pain behavior and the effects of PKCε,TRPV1 and phosphorylation TRPV1(p-TRPV1)in DRG neurons in lumbar‘mild malposition of vertebra and joint’model rats,and to explore the analgesic effect and mechanism of spinal Tuina.Methods:Thirty-two male SD rats were randomly divided into naive group,sham operation group,model group and treatment group,with 8 rats in each group.The‘mild malposition of vertebra and joint’model was prepared by external connection and fixation method.On the 15th day after modeling,the rats in the treatment group were treated with 5 N,2 Hz,10 min/time at L3~S1 Jiaji acupoints on both sides for 14 consecutive days of simulated manipulation.The changes of mechanical pain and thermal pain at different time points were detected in four groups of rats.The dorsal root ganglia were harvested,and the expression and protein content of PKCε,TRPV1 and p-TRPV1 were detected by immunofluorescence and Western Blot techniques.Results:After modeling,the PWT and PWL of the model group and the treatment group were significantly lower than those of the blank group and the sham operation group(P<0.01).On the 10th and 14th days after the intervention,the PWT and PWL of the rats in the treatment group increased significantly than that in the model group(P<0.01).Immunofluorescence showed that the expressions of PKCεand TRPV1 receptors in the DRG of the model group and the treatment group were significantly higher than those of the former two groups,and the expression of PKCεand TRPV1 receptors in the treatment group was significantly inhibited compared with the model group.The protein levels of PKCε,TRPV1,and p-TRPV1 in DRG in the model group and the treatment group were significantly higher than those in the blank and sham operation groups(P<0.01),the protein contents of PKCε,TRPV1 and p-TRPV1 in DRG of the treatment group were lower than those of the model group(P<0.05,P<0.01).Conclusion:Spinal Tuina can improve the rats with low back pain caused by‘mild malposition of verteb

关 键 词：脊柱推拿 椎骨错缝 慢性腰痛 蛋白激酶Cε/瞬时受体电位香草酸亚型1通路 镇痛 磷酸化 机制 

分 类 号：R244.1[医药卫生—针灸推拿学]