优效电针干预不同病理痛的脊髓P2X3机制研究
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摘要
目的:观察优效电针干预对炎性痛和坐骨神经分支选择性损伤大鼠的镇痛作用及脊髓背角P2X3蛋白表达的影响。方法:实验分2部分:1) SD大鼠完全随机分为空白组、CFA组、100 Hz组和假电针组; 2) SD大鼠完全随机分为假SNI组、SNI组、2 Hz组和假电针组。通过足底皮下注射完全弗氏佐剂建立炎性痛模型;通过结扎并切断左侧腓总神经和胫神经后远端,保留腓肠神经的方法建立坐骨神经分支选择性损伤模型。于造模后1 d,100 Hz或2 Hz电针足三里、昆仑穴,持续干预3 d。采用up and down方法检测机械缩足阈,采用免疫印迹法检测患侧脊髓背角P2X3蛋白表达。结果:1)与空白对照组比较,CFA组大鼠机械缩足阈造模后各个时间点都显著下降,CFA组大鼠脊髓背角P2X3蛋白表达增多;与CFA组比较,电针干预1次、3次均能显著提升机械缩足阈,电针干预3次能降低脊髓背角P2X3的蛋白表达,假电针无干预作用。2)与假SNI组比较,SNI组大鼠机械缩足阈造模后各个时间点都显著下降,SNI组大鼠脊髓背角P2X3蛋白表达增多;与SNI组比较,电针干预1次、3次均能显著提升机械缩足阈,电针干预3次能降低脊髓背角P2X3的蛋白表达,假电针无干预作用。结论:100 Hz电针可有效干预慢性炎性痛大鼠机械缩足阈,2 Hz电针可有效干预神经病理痛大鼠机械缩足阈;其镇痛作用可能与下调脊髓背角P2X3蛋白表达相关。
Objective: To observe the analgesic effects of the optimal electroacupuncture intervention on the rats with inflammatory pain and sciatic nerve branch selective injury and the effect of spinal dorsal horn P2 X3 protein expression. Methods: The experiment has two parts. Part I: SD rats were randomly divided into a blank group,a CFA group,a 100 Hz group and a sham electroacupuncture group. Part II: SD rats were completely randomly divided into a sham SNI group,SNI group,2 Hz group and sham electroacupuncture group. The model of inflammatory pain was established by subcutaneous injection of complete freund's adjuvant. By ligating and cutting off the left common peroneal nerve and distal posterior tibial nerve,and preserving the sural nerve,a selective injury model of the sciatic nerve branch was established. At 1 d after molding,100 Hz or 2 Hz electroacupuncture at Zusanli( ST36) and Kunlun( BL60) acupoints continuous intervention for 3 d. the paw withdrawal thresholds was detected by up and down method,and the expression of P2 X3 protein in the dorsal horn of the spinal cord on the affected side was detected by western blot. Results: Part I: Compared with the blank control group,the paw withdrawal thresholds in CFA group decreased significantly at all time points after modeling,while P2 X3 protein expression in spinal dorsal horn of rats in CFA group increased. Compared with CFA group,electroacupuncture intervention once or three times can significantly improve the paw withdrawal thresholds,electroacupuncture intervention three times can reduce the spinal dorsal horn P2 X3 protein expression,and sham electroacupuncture intervention has no effect. Part II: compared with the sham SNI group,the paw withdrawal thresholds were significantly decreased at all time points after modeling,and the P2 X3 protein expression in the spinal dorsal horn of the service group was increased. Compared with the SNI group,1 and 3 times of electroacupuncture intervention could significantly improve the paw withdrawal thresholds,3 times of electroacupuncture intervention could reduce the protein expression of P2 X3 in the spinal dorsal horn,and sham electroacupuncture intervention has no effects. Conclusion: One hundred Hz electroacupuncture can effectively interfere with the paw withdrawal thresholds of chronic inflammatory pain in rats. 2 Hz electroacupuncture can effectively interfere with the paw withdrawal thresholds of neuropathologic pain in rats. Its analgesic effect may be related to down-regulating the expression of P2 X3 protein in spinal dorsal horn.
Objective: To observe the analgesic effects of the optimal electroacupuncture intervention on the rats with inflammatory pain and sciatic nerve branch selective injury and the effect of spinal dorsal horn P2 X3 protein expression. Methods: The experiment has two parts. Part I: SD rats were randomly divided into a blank group,a CFA group,a 100 Hz group and a sham electroacupuncture group. Part II: SD rats were completely randomly divided into a sham SNI group,SNI group,2 Hz group and sham electroacupuncture group. The model of inflammatory pain was established by subcutaneous injection of complete freund's adjuvant. By ligating and cutting off the left common peroneal nerve and distal posterior tibial nerve,and preserving the sural nerve,a selective injury model of the sciatic nerve branch was established. At 1 d after molding,100 Hz or 2 Hz electroacupuncture at Zusanli( ST36) and Kunlun( BL60) acupoints continuous intervention for 3 d. the paw withdrawal thresholds was detected by up and down method,and the expression of P2 X3 protein in the dorsal horn of the spinal cord on the affected side was detected by western blot. Results: Part I: Compared with the blank control group,the paw withdrawal thresholds in CFA group decreased significantly at all time points after modeling,while P2 X3 protein expression in spinal dorsal horn of rats in CFA group increased. Compared with CFA group,electroacupuncture intervention once or three times can significantly improve the paw withdrawal thresholds,electroacupuncture intervention three times can reduce the spinal dorsal horn P2 X3 protein expression,and sham electroacupuncture intervention has no effect. Part II: compared with the sham SNI group,the paw withdrawal thresholds were significantly decreased at all time points after modeling,and the P2 X3 protein expression in the spinal dorsal horn of the service group was increased. Compared with the SNI group,1 and 3 times of electroacupuncture intervention could significantly improve the paw withdrawal thresholds,3 times of electroacupuncture intervention could reduce the protein expression of P2 X3 in the spinal dorsal horn,and sham electroacupuncture intervention has no effects. Conclusion: One hundred Hz electroacupuncture can effectively interfere with the paw withdrawal thresholds of chronic inflammatory pain in rats. 2 Hz electroacupuncture can effectively interfere with the paw withdrawal thresholds of neuropathologic pain in rats. Its analgesic effect may be related to down-regulating the expression of P2 X3 protein in spinal dorsal horn.
引文
[1]陈国良,王梅,路桂军,等.慢性疼痛患者焦虑、抑郁状况调查及相关因素分析[J].中国疼痛医学杂志,2014,20(4):226-230.
[2]王金金,屠文展,蒋松鹤,等.单双侧取穴电针对CCI模型大鼠镇痛效果及脊髓背角P2X3受体表达的影响[J].中华中医药学刊,2014,32(2):305-308.
[3]项璇儿,杜俊英,房军帆,等.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[J].中华中医药杂志,2018,33(5):31-36.
[4]梁宜,周杰,孙晶,等.方剑乔教授电针镇痛临床运用心悟[J].上海针灸杂志,2016,35(12):1387.
[5]周杰,庄晟坚,龚杰,等.不同频率电针对神经病理痛模型大鼠镇痛效应观察[J].浙江中医药大学学报,2016,40(12):887-890.
[6]Fang J Q,Du J Y,Fang J F,et al.Parameter-specific analgesic effects of electroacupuncture mediated by degree of regulation TRPV1 and P2X3 in inflammatory pain in rats[J].Life Sciences,2018,200:69-80.
[7]Chaplan S R,Bach F W,Pogrel J W,et al.Quantitative assessment of tactile allodynia in the rat paw[J].J.Neurosci.Methods,1994,53(1):55-63.
[8]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[J].中华中医药杂志,2012,27(4):957-960.
[9]Isabelle Decosterd,Clifford J Woolf.Spared nerve injury:an animal model of persistent peripheral neuropathic pain[J].Pain,2000,8:3959.
[10]卢峻,时宇静,金智秀,等.不同频率电针对模型大鼠抗抑郁效应的比较研究[J].北京中医药大学学报,2003,26(6):83-85.
[11]Han JS,Wang Q.Mobilization of specific neuropeptides by peripheral stimulation of identified firequencies[J].News in Physiological Sciences,1992,7(4):176-180.
[12]侯丽婷,方先海,刘金锋,等.不同频率电针刺激对神经痛模型大鼠脊髓背角P物质的影响[J].现代生物医学进展,2016,16(12):2267-2270.
[13]孙瑞卿,王贺春,王韵,等.不同频率的电针对大鼠神经源性痛的治疗作用[J].中国应用生理学杂志,2002,18(2):128-131.
[14]蒋袁絮,王韵,刘红香,等.2 Hz和100 Hz经皮神经电刺激对大鼠慢性炎症痛治疗作用的比较[J].中国中西医结合杂志,2001,21(12):923-925.
[15]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[J].浙江中医药大学学报,2016,40(12):891-897,922.
[16]王玉洁,高山,孙肇星,等.舒芬太尼对神经病理性疼痛大鼠脊髓背角P2X3受体表达的影响[J].中国疼痛医学杂志,2017,23(8):575-579.
[17]赵文宇,冷玉芳,周菲,等.青藤碱对神经病理性痛大鼠脊髓背角P2X3R mRNA及COX-2 mRNA表达的影响[J].中华麻醉学杂志,2014,34(9):1098-1100.
[18]Zhang R,Lao L,Ren K,et al.Mechanisms of Acupuncture-Electroacupuncture on Persistent Pain[J].Anesthesiology,2014,120(2):482-503.
[19]李媛,吴凡,程珂,等.针刺对炎性疼痛的镇痛机制[J].针刺研究,2018,43(8):467-475.
[20]Zhao ZQ.Neural mechanism underlying acupuncture analgesia[J].Prog Neurobiol.2008,85(4):355-375.
[21]Zhang R,Lao L,Ren K,Berman BM.Mechanisms of acupunctureelectroacupuncture on persistent pain[J].Anesthesiology,2014,120(2):482-503.
[2]王金金,屠文展,蒋松鹤,等.单双侧取穴电针对CCI模型大鼠镇痛效果及脊髓背角P2X3受体表达的影响[J].中华中医药学刊,2014,32(2):305-308.
[3]项璇儿,杜俊英,房军帆,等.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[J].中华中医药杂志,2018,33(5):31-36.
[4]梁宜,周杰,孙晶,等.方剑乔教授电针镇痛临床运用心悟[J].上海针灸杂志,2016,35(12):1387.
[5]周杰,庄晟坚,龚杰,等.不同频率电针对神经病理痛模型大鼠镇痛效应观察[J].浙江中医药大学学报,2016,40(12):887-890.
[6]Fang J Q,Du J Y,Fang J F,et al.Parameter-specific analgesic effects of electroacupuncture mediated by degree of regulation TRPV1 and P2X3 in inflammatory pain in rats[J].Life Sciences,2018,200:69-80.
[7]Chaplan S R,Bach F W,Pogrel J W,et al.Quantitative assessment of tactile allodynia in the rat paw[J].J.Neurosci.Methods,1994,53(1):55-63.
[8]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[J].中华中医药杂志,2012,27(4):957-960.
[9]Isabelle Decosterd,Clifford J Woolf.Spared nerve injury:an animal model of persistent peripheral neuropathic pain[J].Pain,2000,8:3959.
[10]卢峻,时宇静,金智秀,等.不同频率电针对模型大鼠抗抑郁效应的比较研究[J].北京中医药大学学报,2003,26(6):83-85.
[11]Han JS,Wang Q.Mobilization of specific neuropeptides by peripheral stimulation of identified firequencies[J].News in Physiological Sciences,1992,7(4):176-180.
[12]侯丽婷,方先海,刘金锋,等.不同频率电针刺激对神经痛模型大鼠脊髓背角P物质的影响[J].现代生物医学进展,2016,16(12):2267-2270.
[13]孙瑞卿,王贺春,王韵,等.不同频率的电针对大鼠神经源性痛的治疗作用[J].中国应用生理学杂志,2002,18(2):128-131.
[14]蒋袁絮,王韵,刘红香,等.2 Hz和100 Hz经皮神经电刺激对大鼠慢性炎症痛治疗作用的比较[J].中国中西医结合杂志,2001,21(12):923-925.
[15]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[J].浙江中医药大学学报,2016,40(12):891-897,922.
[16]王玉洁,高山,孙肇星,等.舒芬太尼对神经病理性疼痛大鼠脊髓背角P2X3受体表达的影响[J].中国疼痛医学杂志,2017,23(8):575-579.
[17]赵文宇,冷玉芳,周菲,等.青藤碱对神经病理性痛大鼠脊髓背角P2X3R mRNA及COX-2 mRNA表达的影响[J].中华麻醉学杂志,2014,34(9):1098-1100.
[18]Zhang R,Lao L,Ren K,et al.Mechanisms of Acupuncture-Electroacupuncture on Persistent Pain[J].Anesthesiology,2014,120(2):482-503.
[19]李媛,吴凡,程珂,等.针刺对炎性疼痛的镇痛机制[J].针刺研究,2018,43(8):467-475.
[20]Zhao ZQ.Neural mechanism underlying acupuncture analgesia[J].Prog Neurobiol.2008,85(4):355-375.
[21]Zhang R,Lao L,Ren K,Berman BM.Mechanisms of acupunctureelectroacupuncture on persistent pain[J].Anesthesiology,2014,120(2):482-503.