不同病理痛早期电针镇痛频率优选及其脊髓背角TRPV1活化机制研究
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摘要
目的:筛选炎性痛和神经病理痛早期电针镇痛的优势频率并探讨其脊髓背角TRPV1活化的干预机制。方法:SD大鼠60只,随机分为空白组、炎性痛模型组、2Hz电针Ⅰ组、100Hz电针Ⅰ组、2/100Hz电针Ⅰ组;假模型组、神经病理痛模型组、2Hz电针Ⅱ组、100Hz电针Ⅱ组和2/100Hz电针Ⅱ组,每组6只。炎性痛模型选用弗氏完全佐剂(CFA)模型,神经病理痛模型选用坐骨神经分支选择性损伤(SNI)模型。取双侧足三里、昆仑进行电针干预。采用Von Frey丝检测各组大鼠造模后1d及电针治疗1、3d时患侧50%足底机械缩足反应阈值(PWTs);采用免疫印迹法测大鼠患侧SCDH TRPV1、p-TRPV1的表达。结果:100Hz电针升高CFA大鼠早期患侧PWTs的作用最显著(P<0.05);2Hz电针升高SNI大鼠早期患侧PWTs的效果最显著(P<0.01)。CFA和SNI模型组大鼠早期SCDH TRPV1、p-TRPV1蛋白表达显著升高(P<0.05);100Hz和2Hz电针分别降低了CFA和SNI大鼠早期SCDH TRPV1、p-TRPV1的蛋白表达(P<0.05)。结论:100Hz、2Hz电针分别为干预炎性痛和神经病理痛早期的优势频率电针;优势频率电针的镇痛效应可能通过调控炎性痛和神经病理痛大鼠早期SCDH TRPV1受体活化实现。
Objective: To select the optimal frequency of electroacupuncture(EA) analgesia in inflammatory pain and neuropathic pain and explore the interventional mechanism of TRPV1 activation in the spinal cord dorsal horn. Methods: Sixty SD rats were randomly divided into blank group, inflammatory pain model group, 2 Hz EA group Ⅰ, 100 Hz EA group Ⅱ, 2/100 Hz EA group Ⅰ; sham model group, neuropathic pain model group, 2 Hz EA group Ⅰ, 100 Hz EA group Ⅱand 2/100 Hz EA groupⅡ, n=6. Freund's complete adjuvant(CFA) model and spared nerve injury(SNI) model were established as inflammatory pain model and neuropathic pain model respectively. EA was administered at bilateral acupoints Zusanli(ST36) and Kunlun(BL60). The affected side 50% paw withdrawal thresholds(PWTs) were detected by Von Frey hairs at time points of 1 day after modelestablishment, 1 day and 3 days after EA intervention in each group. The levels of TRPV1, p-TRPV1 in affected side SCDH were tested by Western blot. Results: Affected side PWTs of CFA rats in 100 Hz EA group Ⅰ were increased most obviously in EA groups(P<0.05), while affected side PWTs of SNI rats in 2 Hz EA group Ⅱ were increased most obviously in EA groups(P<0.01). The expression of TRPV1 and p-TRPV1 protein in SCDH were significantly increased in the early phase of both CFA and SNI rats(P<0.05); but 100 Hz EA and 2 Hz EA decreased the expression of SCDH TRPV1, p-TRPV1 level of CFA and SNI rats respectively(P<0.05). Conclusion: 100 Hz and 2 Hz are the optimal frequency of EA for early inflammatory pain and neuropathic pain. The analgesic effect of the optimal frequency EA may be accomplished by regulating the activation of SCDH TRPV1 receptor in early inflammatory pain and neuropathic pain models.
Objective: To select the optimal frequency of electroacupuncture(EA) analgesia in inflammatory pain and neuropathic pain and explore the interventional mechanism of TRPV1 activation in the spinal cord dorsal horn. Methods: Sixty SD rats were randomly divided into blank group, inflammatory pain model group, 2 Hz EA group Ⅰ, 100 Hz EA group Ⅱ, 2/100 Hz EA group Ⅰ; sham model group, neuropathic pain model group, 2 Hz EA group Ⅰ, 100 Hz EA group Ⅱand 2/100 Hz EA groupⅡ, n=6. Freund's complete adjuvant(CFA) model and spared nerve injury(SNI) model were established as inflammatory pain model and neuropathic pain model respectively. EA was administered at bilateral acupoints Zusanli(ST36) and Kunlun(BL60). The affected side 50% paw withdrawal thresholds(PWTs) were detected by Von Frey hairs at time points of 1 day after modelestablishment, 1 day and 3 days after EA intervention in each group. The levels of TRPV1, p-TRPV1 in affected side SCDH were tested by Western blot. Results: Affected side PWTs of CFA rats in 100 Hz EA group Ⅰ were increased most obviously in EA groups(P<0.05), while affected side PWTs of SNI rats in 2 Hz EA group Ⅱ were increased most obviously in EA groups(P<0.01). The expression of TRPV1 and p-TRPV1 protein in SCDH were significantly increased in the early phase of both CFA and SNI rats(P<0.05); but 100 Hz EA and 2 Hz EA decreased the expression of SCDH TRPV1, p-TRPV1 level of CFA and SNI rats respectively(P<0.05). Conclusion: 100 Hz and 2 Hz are the optimal frequency of EA for early inflammatory pain and neuropathic pain. The analgesic effect of the optimal frequency EA may be accomplished by regulating the activation of SCDH TRPV1 receptor in early inflammatory pain and neuropathic pain models.
引文
[1]Johannes C B,Le T K,Zhou X,et al.The prevalence of chronic pain in United States adults:results of an Internet-based survey.Journal of Pain,2010,11(11):1230-1239
[2]韩济生.疼痛学.北京大学医学出版社,2012:4
[3]Honda K,Shinoda M,Kondo M,et al.Sensitization of TRPV1 and TRPA1 via peripheral m Glu R5 signaling contributes to thermal and mechanical hypersensitivity.Pain,2017,158(9):1754
[4]Choi S I,Lim J Y,Yoo S,et al.Emerging Role of Spinal Cord TRPV1in Pain Exacerbation.Neural Plasticity,2016,2016(2):5954890
[5]Green D,Ruparel S,Gao X,et al.Central activation of TRPV1 and TRPA1 by novel endogenous agonists contributes to mechanical allodynia and thermal hyperalgesia after burn injury.Molecular Pain,2016,12:1-9
[6]崔东,李泽华,宋学军.慢性疼痛的脊髓机制.中国疼痛医学杂志,2017,23(9):641-647
[7]Lappin S C,Randall A D,Gunthorpe M J,et al.TRPV1 antagonist,SB-366791,inhibits glutamatergic synaptic transmission in rat spinal dorsal horn following peripheral inflammation.European Journal of Pharmacology,2006,540(1-3):73-81
[8]Kalynovska N,Adamek P,Palecek J.TRPV1 receptors contribute to mediate paclitaxel-induced c-Fos expression in spinal cord dorsal horn neurons.Physiological Research,2017,66(3):549-552
[9]刘盈君,林小溪,方芳,等.Mas相关G蛋白偶联受体C参与电针干预慢性炎性痛及其外周δ-阿片受体机制.中华中医药杂志,2017,32(5):1988-1993
[10]项璇儿,杜俊英,方剑乔,等.电针对CFA致炎性痛大鼠热痛和机械痛优效频率观察及机制初探.浙江中医药大学学报,2016,40(12):891-897,922
[11]叶佳瑜,蒋永亮,颜思思,等.低频电针对选择性神经损伤大鼠神经痛维持期脊髓背角PKA-TRPV1通路及痛敏递质的干预作用.中华物理医学与康复杂志,2017,39(7):481-485
[12]华兴邦,周浩良.大鼠穴位图谱的研制.实验动物与动物实验,1991(1):1-5
[13]Kurganova Y M,Danilov A B.Melatonin in Chronic Pain Syndromes.Neuroscience&Behavioral Physiology,2017,47(7):806-812
[14]张梦.疼痛相关疾病动物模型研究概况.西部医学,2014,26(6):814-816
[15]Beyer A,Sch?fer M,Stein C.Antinociceptive effects of dynorphin peptides in a model of inflammatory pain.PAIN,1997,70(2):141-147
[16]杜俊英,方剑乔,梁宜,等.电针对CFA致炎性痛大鼠脊髓ATF-2Thr71位点磷酸化的干预研究.中华中医药杂志,2012,27(4):957-960
[17]M Lukács,K Warfvinge,J Tajti,et al.Topical dura mater application of CFA induces enhanced expression of c-fos and glutamate in rat trigeminal nucleus caudalis:attenuated by KYNA derivate(SZR72).Journal of Headache&Pain,2017,18(1):39-46
[18]叶佳瑜,颜思思,蒋永亮,等.4种常用神经病理痛动物模型的评价.云南中医学院学报,2015,38(4):92-95
[19]周杰,陈贞羽,龚杰,等.不同参数组合电针对SNL大鼠镇痛效应观察及对下丘脑内啡肽的影响.中国医药导报,2017,14(23):18-21
[20]刘盈君,方芳,方剑乔,等.电针对慢性炎性痛大鼠镇痛作用及机制研究.中国中西医结合杂志,2016,36(6):690-695
[21]周杰,陈贞羽,龚杰,等.不同参数组合电针对SNL大鼠镇痛效应观察及对脊髓背角PGE2的影响.中国现代医生,2017,55(24):33-36
[22]张瑛,王韵.TRPV1:一种同时参与慢性痛外周敏化和疼痛中枢调制的重要分子.生理学报,2017(5):1-16
[23]Abooj M,Bishnoi M,Bosgraaf C A,et al.Changes in spinal cord following inflammatory and neuropathic pain and the effectiveness of resiniferatoxin.Open Pain Journal,2016,9(1):1-14
[24]颜思思,蒋永亮,叶佳瑜,等.SNI大鼠神经痛维持期脊髓背角TRPV1的活化形式及低频电针干预作用.浙江中医药大学学报,2016,40(5):323-329,352
[25]蒋永亮,尹小虎,沈亚芳,等.大鼠SNL神经痛模型不同时相背根神经节TRPV1的活化形式.中国疼痛医学杂志,2014,20(6):383-387
[26]Xiao X,Zhao X T,Xu L C,et al.Shp-1 dephosphorylates TRPV1in dorsal root ganglion neurons and alleviates CFA-induced inflammatory pain in rats.Pain,2015,156(4):597-608
[27]Liao H Y,Hsieh C L,Huang C P,et al.Electroacupuncture attenuates CFA-induced inflammatory pain by suppressing Nav1.8 through S100B,TRPV1,Opioid,and Adenosine pathways in mice.Scientific Reports,2017,7:42531
[28]Wu C H,Lv Z T,Zhao Y,et al.Electroacupuncture improves thermal and mechanical sensitivities in a rat model of postherpetic neuralgia.Molecular Pain,2013,9(1):18-27
[29]蒋永亮,沈亚芳,尹小虎,等.低频电针对SNL大鼠早期神经痛DRG p-TRPV1、CGRP的抑制作用.浙江中医药大学学报,2014,38(6):780-784
[2]韩济生.疼痛学.北京大学医学出版社,2012:4
[3]Honda K,Shinoda M,Kondo M,et al.Sensitization of TRPV1 and TRPA1 via peripheral m Glu R5 signaling contributes to thermal and mechanical hypersensitivity.Pain,2017,158(9):1754
[4]Choi S I,Lim J Y,Yoo S,et al.Emerging Role of Spinal Cord TRPV1in Pain Exacerbation.Neural Plasticity,2016,2016(2):5954890
[5]Green D,Ruparel S,Gao X,et al.Central activation of TRPV1 and TRPA1 by novel endogenous agonists contributes to mechanical allodynia and thermal hyperalgesia after burn injury.Molecular Pain,2016,12:1-9
[6]崔东,李泽华,宋学军.慢性疼痛的脊髓机制.中国疼痛医学杂志,2017,23(9):641-647
[7]Lappin S C,Randall A D,Gunthorpe M J,et al.TRPV1 antagonist,SB-366791,inhibits glutamatergic synaptic transmission in rat spinal dorsal horn following peripheral inflammation.European Journal of Pharmacology,2006,540(1-3):73-81
[8]Kalynovska N,Adamek P,Palecek J.TRPV1 receptors contribute to mediate paclitaxel-induced c-Fos expression in spinal cord dorsal horn neurons.Physiological Research,2017,66(3):549-552
[9]刘盈君,林小溪,方芳,等.Mas相关G蛋白偶联受体C参与电针干预慢性炎性痛及其外周δ-阿片受体机制.中华中医药杂志,2017,32(5):1988-1993
[10]项璇儿,杜俊英,方剑乔,等.电针对CFA致炎性痛大鼠热痛和机械痛优效频率观察及机制初探.浙江中医药大学学报,2016,40(12):891-897,922
[11]叶佳瑜,蒋永亮,颜思思,等.低频电针对选择性神经损伤大鼠神经痛维持期脊髓背角PKA-TRPV1通路及痛敏递质的干预作用.中华物理医学与康复杂志,2017,39(7):481-485
[12]华兴邦,周浩良.大鼠穴位图谱的研制.实验动物与动物实验,1991(1):1-5
[13]Kurganova Y M,Danilov A B.Melatonin in Chronic Pain Syndromes.Neuroscience&Behavioral Physiology,2017,47(7):806-812
[14]张梦.疼痛相关疾病动物模型研究概况.西部医学,2014,26(6):814-816
[15]Beyer A,Sch?fer M,Stein C.Antinociceptive effects of dynorphin peptides in a model of inflammatory pain.PAIN,1997,70(2):141-147
[16]杜俊英,方剑乔,梁宜,等.电针对CFA致炎性痛大鼠脊髓ATF-2Thr71位点磷酸化的干预研究.中华中医药杂志,2012,27(4):957-960
[17]M Lukács,K Warfvinge,J Tajti,et al.Topical dura mater application of CFA induces enhanced expression of c-fos and glutamate in rat trigeminal nucleus caudalis:attenuated by KYNA derivate(SZR72).Journal of Headache&Pain,2017,18(1):39-46
[18]叶佳瑜,颜思思,蒋永亮,等.4种常用神经病理痛动物模型的评价.云南中医学院学报,2015,38(4):92-95
[19]周杰,陈贞羽,龚杰,等.不同参数组合电针对SNL大鼠镇痛效应观察及对下丘脑内啡肽的影响.中国医药导报,2017,14(23):18-21
[20]刘盈君,方芳,方剑乔,等.电针对慢性炎性痛大鼠镇痛作用及机制研究.中国中西医结合杂志,2016,36(6):690-695
[21]周杰,陈贞羽,龚杰,等.不同参数组合电针对SNL大鼠镇痛效应观察及对脊髓背角PGE2的影响.中国现代医生,2017,55(24):33-36
[22]张瑛,王韵.TRPV1:一种同时参与慢性痛外周敏化和疼痛中枢调制的重要分子.生理学报,2017(5):1-16
[23]Abooj M,Bishnoi M,Bosgraaf C A,et al.Changes in spinal cord following inflammatory and neuropathic pain and the effectiveness of resiniferatoxin.Open Pain Journal,2016,9(1):1-14
[24]颜思思,蒋永亮,叶佳瑜,等.SNI大鼠神经痛维持期脊髓背角TRPV1的活化形式及低频电针干预作用.浙江中医药大学学报,2016,40(5):323-329,352
[25]蒋永亮,尹小虎,沈亚芳,等.大鼠SNL神经痛模型不同时相背根神经节TRPV1的活化形式.中国疼痛医学杂志,2014,20(6):383-387
[26]Xiao X,Zhao X T,Xu L C,et al.Shp-1 dephosphorylates TRPV1in dorsal root ganglion neurons and alleviates CFA-induced inflammatory pain in rats.Pain,2015,156(4):597-608
[27]Liao H Y,Hsieh C L,Huang C P,et al.Electroacupuncture attenuates CFA-induced inflammatory pain by suppressing Nav1.8 through S100B,TRPV1,Opioid,and Adenosine pathways in mice.Scientific Reports,2017,7:42531
[28]Wu C H,Lv Z T,Zhao Y,et al.Electroacupuncture improves thermal and mechanical sensitivities in a rat model of postherpetic neuralgia.Molecular Pain,2013,9(1):18-27
[29]蒋永亮,沈亚芳,尹小虎,等.低频电针对SNL大鼠早期神经痛DRG p-TRPV1、CGRP的抑制作用.浙江中医药大学学报,2014,38(6):780-784