电针对结直肠扩张刺激模型大鼠内脏伤害性痛反应的影响
|
摘要
目的探讨不同强度、不同穴位电针刺激对结直肠扩张刺激引起的大鼠内脏伤害性痛反应的影响及其可能机制。方法 SD大鼠54只,分别进行电生理实验(30只)和形态学实验(24只)。电生理实验中先选出6只大鼠用于结直肠扩张(CRD)刺激诱导腹直肌肌电发生阈值观察,其余大鼠随机分为足三里组、天枢组和曲池组,每组8只,麻醉状态下给予CRD刺激造成大鼠内脏伤害性疼痛,采用1 mA和3 mA电流强度电针刺激大鼠"足三里""天枢""曲池",持续时间140 s,记录其对腹直肌肌电活动的曲线下面积、频率、振幅。形态学实验大鼠随机分为正常组、模型组、1 mA足三里组、1 mA天枢组、1 mA曲池组、3 mA足三里组、3 mA天枢组、3 mA曲池组,每组3只,电针刺激持续时间30 min,采用免疫组化方法观察不同强度不同穴位电针刺激对CRD刺激造成孤束核c-fos阳性神经元表达的影响。结果 60 mmHg的CRD可诱发大鼠内脏痛,表现为腹直肌肌电的曲线下面积、频率和振幅明显增加,与CRD前相比差异有统计学意义(P <0. 01),故将其选为CRD刺激值。1 mA电针"天枢"可以显著改善伤害性刺激引起的腹直肌肌电的发放(P <0. 05),c-fos阳性神经元在延髓孤束核表达的数量和密度均明显减少(P <0. 05)。而1 mA电针"足三里"和"曲池",对CRD引起的腹直肌肌电发放没有显著改善。3 mA电针"足三里"和"天枢",均可以明显抑制CRD引起的腹直肌肌电的发放(P <0. 01),并减少CRD造成c-fos在孤束核的表达增加(P <0. 05)。结论电针"天枢"和"足三里"均可以显著改善CRD引起的伤害性痛反应,减少c-fos阳性神经元在大鼠延髓孤束核的表达,但同节段"天枢"抑制内脏痛所需的刺激量较小。
Objective To discuss the effects of different intensity electroacupuncture( EA) at different acupoints on visceral nociceptive pain response induced by colorectal distention( CRD) and its possible mechanism. Methods A total of 54 Sprague-Dawley( SD) rats were received electrophysiological experiments( 30 rats) and morphological experiments( 24 rats). In electrophysiological experiments,6 rats were selected for CRD stimulation to induce the rectus abdominis myoelectricity threshold. The remaining rats were randomly divided into Zusanli( ST36) group,Tianshu( ST25) group and Quchi( LI11) group,with 8 rats in each group. Under inhalation anesthesia with isoflurane,CRD stimulation was given to cause visceral injury pain. EA with current of 1 mA and 3 mA were used to stimulate rats' "Zusanli"( ST36),"Tianshu"( ST25),and "Quchi"( LI11),for 140 s. The area,frequency,and amplitude of the curve of the rectus abdominis myoelectric activity were recorded. The rats for morphological experiments were randomly divided into normal group,model group,1 mA Zusanli( ST36) group,1 mA Tianshu( ST25) group,1 mA Quchi( LI11) group,3 mA Zusanli( ST36) group,3 mA Tianshu( ST25) group and 3 mA Quchi( LI11)group,with 3 rats in each group. EA stimulation last for 30 min. Immunohistochemical method was used to observe the effects of EA stimulation with different intensity at different acupoints on the expression of c-fos positive neurons in solitary tract nucleus. Results CRD of 60 mmHg could induce visceral pain of rats,which showed that the area,frequency and amplitude of the rectus abdominis myoelectric curve increased significantly,and the difference was statistically significant than before CRD stimulation( P < 0. 01),so it was selected as CRD stimulus value. 1 mA EA at"Tianshu"( ST25) could significantly improve the release of rectus abdominis myoelectricity caused by noxious stimulation( P < 0. 05),and the number and density of c-fos positive neurons in the nucleus tractus solitarii were significantly decreased( P < 0. 05). The 1 mA EA at"Zusanli"( ST36) and"Quchi"( LI11) did not significantly improve the rectus abdominis myoelectrical delivery caused by CRD. 3 mA EA at "Zusanli"( ST36) and "Tianshu"( ST25)significantly inhibited the activity of EMG induced by CRD( P < 0. 01),and also significantly reduced the number and fluorescence intensity of c-fos positive neurons in nucleus tractus solitarii( P < 0. 05). Conclusion EA at"Tianshu"( ST25) and"Zusanli"( ST36) significantly inhibited the nociceptive response induced by CRD,and decreased the expression of c-fos positive neurons in nucleus tractus solitarii of rats. In the same segment of the spinal cord,EA at "Tianshu"( ST25) needed less intensity to produce inhibitory effects.
Objective To discuss the effects of different intensity electroacupuncture( EA) at different acupoints on visceral nociceptive pain response induced by colorectal distention( CRD) and its possible mechanism. Methods A total of 54 Sprague-Dawley( SD) rats were received electrophysiological experiments( 30 rats) and morphological experiments( 24 rats). In electrophysiological experiments,6 rats were selected for CRD stimulation to induce the rectus abdominis myoelectricity threshold. The remaining rats were randomly divided into Zusanli( ST36) group,Tianshu( ST25) group and Quchi( LI11) group,with 8 rats in each group. Under inhalation anesthesia with isoflurane,CRD stimulation was given to cause visceral injury pain. EA with current of 1 mA and 3 mA were used to stimulate rats' "Zusanli"( ST36),"Tianshu"( ST25),and "Quchi"( LI11),for 140 s. The area,frequency,and amplitude of the curve of the rectus abdominis myoelectric activity were recorded. The rats for morphological experiments were randomly divided into normal group,model group,1 mA Zusanli( ST36) group,1 mA Tianshu( ST25) group,1 mA Quchi( LI11) group,3 mA Zusanli( ST36) group,3 mA Tianshu( ST25) group and 3 mA Quchi( LI11)group,with 3 rats in each group. EA stimulation last for 30 min. Immunohistochemical method was used to observe the effects of EA stimulation with different intensity at different acupoints on the expression of c-fos positive neurons in solitary tract nucleus. Results CRD of 60 mmHg could induce visceral pain of rats,which showed that the area,frequency and amplitude of the rectus abdominis myoelectric curve increased significantly,and the difference was statistically significant than before CRD stimulation( P < 0. 01),so it was selected as CRD stimulus value. 1 mA EA at"Tianshu"( ST25) could significantly improve the release of rectus abdominis myoelectricity caused by noxious stimulation( P < 0. 05),and the number and density of c-fos positive neurons in the nucleus tractus solitarii were significantly decreased( P < 0. 05). The 1 mA EA at"Zusanli"( ST36) and"Quchi"( LI11) did not significantly improve the rectus abdominis myoelectrical delivery caused by CRD. 3 mA EA at "Zusanli"( ST36) and "Tianshu"( ST25)significantly inhibited the activity of EMG induced by CRD( P < 0. 01),and also significantly reduced the number and fluorescence intensity of c-fos positive neurons in nucleus tractus solitarii( P < 0. 05). Conclusion EA at"Tianshu"( ST25) and"Zusanli"( ST36) significantly inhibited the nociceptive response induced by CRD,and decreased the expression of c-fos positive neurons in nucleus tractus solitarii of rats. In the same segment of the spinal cord,EA at "Tianshu"( ST25) needed less intensity to produce inhibitory effects.
引文
[1]GIAMBERARDINO MA.Recent and forgotten aspects of visceral pain[J].Eur J Pain,1999,3(2):77-92.
[2]CAMILLERI M,BOECKXSTAENS G.Dietary and pharmacological treatment of abdominal pain in IBS[J].Gut,2017,66(5):966-974.
[3]王美青,潘寅兵,钱燕宁.腹腔镜胆囊切除术后疼痛的原因及对策[J].临床麻醉学杂志,2013,29(11):1126-1129.
[4]刘春亮,熊源长,卢军,等.腕踝针治疗腹腔镜胆囊术后疼痛疗效观察[J].上海针灸杂志,2016,35(3):297-300.
[5]李芳.艾灸疗法在肠腑病内脏痛中镇痛的效应及作用机制研究[J].中医临床研究,2014,6(21):113-114.
[6]XING J,LARIVE B,MEKHAIL N,et al.Transcutaneous electrical acustimulation can reduce visceral perception in patients with the irritable bowel syndrome:a pilot study[J].Altern Ther Health Med,2004,10(1):38-42.
[7]XIAO WB,LIU YL.Rectal hypersensitivity reduced by acupoint TENS in patients with diarrhea-predominant irritable bowel syndrome:a pilot study[J].Dig Dis Sci,2004,49(2):312-319.
[8]李为民,吴根诚.电针对急性内脏痛大鼠腹部撤回反射和腹直肌肌电的影响[J].中国临床康复,2004,8(10):1899-1901.
[9]周娟,李为民.针刺治疗内脏痛的基础研究进展[J].上海针灸杂志,2007,26(10):45-48.
[10]吴璐一,翁志军,窦传字,等.针灸对IBS内脏痛大鼠结肠肌间神经丛神经元P2X2受体表达的调节[J].世界中西医结合杂志,2014,9(6):601-604.
[11]付宏伟,阎丽娟,刘阳阳,等.不同电针刺激参数对镇痛效应影响的研究概述[J].上海针灸杂志,2018,37(11):1331-1335.
[12]王友京,王双坤.不同强度和频率电针的镇痛效应[J].针刺研究,1993,18(1):44-47.
[13]王珊,高誉珊,蓝莹,等.电针“内关”“天枢”穴对肠易激综合征大鼠结肠动力及结肠D2受体的影响[J].针刺研究,2018,43(1):24-28.
[14]杨建平.大鼠结直肠扩张内脏痛模型的建立与行为学评价[J].中华实验外科杂志,2003,20(12):62-64.
[15]ZHOU YY,WANNER NJ,XIAO Y,et al.Electroacupuncture alleviates stress-induced visceral hypersensitivity through an opioid system in rats[J].World J Gastroenterol,2012,18(48):7201-7211.
[16]张露芬.实验针灸学[M].北京:化学工业出版社,2010:219-221.
[17]吴红金,薛峥,茹立强.电针抗大鼠急性炎症性内脏痛的肠神经机制[J].针刺研究,1999,24(2):138-142.
[18]XIN J,SU Y,YANG Z,et al.Distinct roles of ASIC3and TRPV1 receptors in electroacupuncture-induced segmental and systemic analgesia[J].Front Med,2016,10(4):465-472.
[19]KEAST JR,DE GROAT WC.Segmental distribution and peptide content of primary afferent neurons innervating the urogenital organs and colon of male rats[J].J Comp Neurol,1992,319(4):615-623.
[20]QIAO LY,GRIDER JR.Up-regulation of calcitonin gene-related peptide and receptor tyrosine kinase TrkB in rat bladder afferent neurons following TNBS colitis[J].Exp Neurol,2007,204(2):667-679.
[21]ZHU B,XU WD,RONG PJ,et al.A C-fiber reflex inhibition induced by electroacupuncture with different intensities applied at homotopic and heterotopic acupoints in rats selectively destructive effects on myelinated and unmyelinated afferent fibers[J].Brain Res,2004,1011(2):228-237.
[22]刘乡.以痛制痛:针刺镇痛的基本神经机制[J].科学通报,2001,46(7):609-616.
[23]CUI JJ,ZHU XL,SHI H,et al.The expression of calcitonin gene-related peptide on the neurons associated Zusanli(ST 36)in rats[J].Chin J Integr Med,2015,21(8):630-634.
[24]余玲玲,李亮,秦庆广,等.内脏伤害性传入易化穴位对大鼠脊髓广动力型神经元的激活效应[J].针刺研究,2014,39(5):390-395.
[25]朱丽霞,李文武,吉长福,等.不同强度电针对痛觉异常治疗作用的比较[J].中国疼痛医学杂志,1996,2(1):26-32.
[26]辛娟娟,宿杨帅,杨兆坤,等.不同强度电针、热灸样刺激对香草酸瞬时受体亚型1基因敲除小鼠痛阈的影响[J].针刺研究,2012,37(6):431-439.
[27]LEE CH,JUNG HS,LEE TY,et al.Studies of the central neural pathways to the stomach and Zusanli(ST36)[J].Am J Chin Med,2001,29(2):211-220.
[28]TANIDA M,TAO Z,SUN L,et al.Anaphylactic hypotension causes renal and adrenal sympathoexcitaion and induces c-fos in the hypothalamus and medulla oblongata:Anaphylaxis affects the brain and activates sympathetic nerves[J].Exp Physiol,2018,103(6):790-806.
[29]STAM R,EKKELENKAMP K,FRANKHUIJZEN AC,et al.Long-lasting changes in central nervous system responsivity to colonic distention after stress in rats[J].Gastroenterology,2002,123(4):1216-1225.
[30]OKUSHA Y,HIRAI Y,MAEZAWA H,et al.Effects of intraperitoneally administered L-histidine on food intake,taste,and visceral sensation in rats[J].J Physiol Sci,2017,67(4):467-474.
[31]陈良为,晋光荣,饶志仁,等.大鼠胃肠道伤害性刺激引起的中枢神经系统c-fos表达[J].神经解剖学杂志,1993,9(2):185-191,264.
[32]秦明,黄裕新,王景杰,等.电针“足三里”穴对内脏痛大鼠延髓内脏带内c-fos和GFAP表达的影响[J].针刺研究,2006,31(3):136-139,148,193.
[33]易春霞,胡道松.电针抗胃炎性痛对内脏感觉c-fos及NOS阳性神经元的影响[J].针刺研究,2004,29(1):24-30.
[34]王伟东,岳柏林,孔抗美,等.电针刺激曲池穴在颈脊髓效应的功能磁共振成像研究[J].中国实用医药,2014,9(29):1-2.
[35]齐琳婧,苏同生,刘志顺,等.针刺曲池上巨虚治疗功能性肠病临床观察[J].陕西中医,2014,35(2):224-225.
[36]盛蓉蓉,章薇.曲池穴的临床应用和实验研究进展[J].中国医药指南,2013,11(20):81-82.
[2]CAMILLERI M,BOECKXSTAENS G.Dietary and pharmacological treatment of abdominal pain in IBS[J].Gut,2017,66(5):966-974.
[3]王美青,潘寅兵,钱燕宁.腹腔镜胆囊切除术后疼痛的原因及对策[J].临床麻醉学杂志,2013,29(11):1126-1129.
[4]刘春亮,熊源长,卢军,等.腕踝针治疗腹腔镜胆囊术后疼痛疗效观察[J].上海针灸杂志,2016,35(3):297-300.
[5]李芳.艾灸疗法在肠腑病内脏痛中镇痛的效应及作用机制研究[J].中医临床研究,2014,6(21):113-114.
[6]XING J,LARIVE B,MEKHAIL N,et al.Transcutaneous electrical acustimulation can reduce visceral perception in patients with the irritable bowel syndrome:a pilot study[J].Altern Ther Health Med,2004,10(1):38-42.
[7]XIAO WB,LIU YL.Rectal hypersensitivity reduced by acupoint TENS in patients with diarrhea-predominant irritable bowel syndrome:a pilot study[J].Dig Dis Sci,2004,49(2):312-319.
[8]李为民,吴根诚.电针对急性内脏痛大鼠腹部撤回反射和腹直肌肌电的影响[J].中国临床康复,2004,8(10):1899-1901.
[9]周娟,李为民.针刺治疗内脏痛的基础研究进展[J].上海针灸杂志,2007,26(10):45-48.
[10]吴璐一,翁志军,窦传字,等.针灸对IBS内脏痛大鼠结肠肌间神经丛神经元P2X2受体表达的调节[J].世界中西医结合杂志,2014,9(6):601-604.
[11]付宏伟,阎丽娟,刘阳阳,等.不同电针刺激参数对镇痛效应影响的研究概述[J].上海针灸杂志,2018,37(11):1331-1335.
[12]王友京,王双坤.不同强度和频率电针的镇痛效应[J].针刺研究,1993,18(1):44-47.
[13]王珊,高誉珊,蓝莹,等.电针“内关”“天枢”穴对肠易激综合征大鼠结肠动力及结肠D2受体的影响[J].针刺研究,2018,43(1):24-28.
[14]杨建平.大鼠结直肠扩张内脏痛模型的建立与行为学评价[J].中华实验外科杂志,2003,20(12):62-64.
[15]ZHOU YY,WANNER NJ,XIAO Y,et al.Electroacupuncture alleviates stress-induced visceral hypersensitivity through an opioid system in rats[J].World J Gastroenterol,2012,18(48):7201-7211.
[16]张露芬.实验针灸学[M].北京:化学工业出版社,2010:219-221.
[17]吴红金,薛峥,茹立强.电针抗大鼠急性炎症性内脏痛的肠神经机制[J].针刺研究,1999,24(2):138-142.
[18]XIN J,SU Y,YANG Z,et al.Distinct roles of ASIC3and TRPV1 receptors in electroacupuncture-induced segmental and systemic analgesia[J].Front Med,2016,10(4):465-472.
[19]KEAST JR,DE GROAT WC.Segmental distribution and peptide content of primary afferent neurons innervating the urogenital organs and colon of male rats[J].J Comp Neurol,1992,319(4):615-623.
[20]QIAO LY,GRIDER JR.Up-regulation of calcitonin gene-related peptide and receptor tyrosine kinase TrkB in rat bladder afferent neurons following TNBS colitis[J].Exp Neurol,2007,204(2):667-679.
[21]ZHU B,XU WD,RONG PJ,et al.A C-fiber reflex inhibition induced by electroacupuncture with different intensities applied at homotopic and heterotopic acupoints in rats selectively destructive effects on myelinated and unmyelinated afferent fibers[J].Brain Res,2004,1011(2):228-237.
[22]刘乡.以痛制痛:针刺镇痛的基本神经机制[J].科学通报,2001,46(7):609-616.
[23]CUI JJ,ZHU XL,SHI H,et al.The expression of calcitonin gene-related peptide on the neurons associated Zusanli(ST 36)in rats[J].Chin J Integr Med,2015,21(8):630-634.
[24]余玲玲,李亮,秦庆广,等.内脏伤害性传入易化穴位对大鼠脊髓广动力型神经元的激活效应[J].针刺研究,2014,39(5):390-395.
[25]朱丽霞,李文武,吉长福,等.不同强度电针对痛觉异常治疗作用的比较[J].中国疼痛医学杂志,1996,2(1):26-32.
[26]辛娟娟,宿杨帅,杨兆坤,等.不同强度电针、热灸样刺激对香草酸瞬时受体亚型1基因敲除小鼠痛阈的影响[J].针刺研究,2012,37(6):431-439.
[27]LEE CH,JUNG HS,LEE TY,et al.Studies of the central neural pathways to the stomach and Zusanli(ST36)[J].Am J Chin Med,2001,29(2):211-220.
[28]TANIDA M,TAO Z,SUN L,et al.Anaphylactic hypotension causes renal and adrenal sympathoexcitaion and induces c-fos in the hypothalamus and medulla oblongata:Anaphylaxis affects the brain and activates sympathetic nerves[J].Exp Physiol,2018,103(6):790-806.
[29]STAM R,EKKELENKAMP K,FRANKHUIJZEN AC,et al.Long-lasting changes in central nervous system responsivity to colonic distention after stress in rats[J].Gastroenterology,2002,123(4):1216-1225.
[30]OKUSHA Y,HIRAI Y,MAEZAWA H,et al.Effects of intraperitoneally administered L-histidine on food intake,taste,and visceral sensation in rats[J].J Physiol Sci,2017,67(4):467-474.
[31]陈良为,晋光荣,饶志仁,等.大鼠胃肠道伤害性刺激引起的中枢神经系统c-fos表达[J].神经解剖学杂志,1993,9(2):185-191,264.
[32]秦明,黄裕新,王景杰,等.电针“足三里”穴对内脏痛大鼠延髓内脏带内c-fos和GFAP表达的影响[J].针刺研究,2006,31(3):136-139,148,193.
[33]易春霞,胡道松.电针抗胃炎性痛对内脏感觉c-fos及NOS阳性神经元的影响[J].针刺研究,2004,29(1):24-30.
[34]王伟东,岳柏林,孔抗美,等.电针刺激曲池穴在颈脊髓效应的功能磁共振成像研究[J].中国实用医药,2014,9(29):1-2.
[35]齐琳婧,苏同生,刘志顺,等.针刺曲池上巨虚治疗功能性肠病临床观察[J].陕西中医,2014,35(2):224-225.
[36]盛蓉蓉,章薇.曲池穴的临床应用和实验研究进展[J].中国医药指南,2013,11(20):81-82.