电针“天枢”“大肠俞”穴对肠易激综合征大鼠肠道动力和敏感性影响的比较研究
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摘要
目的:观察电针"天枢"和"大肠俞"对肠易激综合征(IBS)大鼠的肠道压力值和敏感性的影响,并探讨其效应差异及相关机制。方法:新生Wistar大鼠40只,按随机数字表法分为空白对照组、模型对照组、天枢组和大肠俞组,每组10只。采用新生期母子分离、醋酸灌肠和结直肠扩张联合制备IBS模型。9周鼠龄时,天枢组和大肠俞组分别对双侧"天枢"和"大肠俞"进行电针干预,隔日1次,每次20min,共5次。通过改良的腹部回撤反射法评估肠道敏感性和动力,即记录肠道起始收缩波潜伏期、90s内收缩波个数、90s内收缩波峰峰值;采用免疫组化法检测结肠肌层M3受体及黏膜层5-羟色胺(HT)3A受体的阳性表达。结果:与空白对照组相比,模型对照组肠道起始波潜伏期缩短,90s内收缩波个数增多,90s内收缩波峰峰值增大(P<0.01),M3和5-HT3A受体阳性表达增强(P<0.01)。与模型对照组相比,天枢组和大肠俞组潜伏期延长、峰峰值变小(P<0.01),天枢组收缩波个数减少(P<0.01),天枢组和大肠俞组M3和5-HT3A受体阳性表达减少(P<0.01)。与天枢组相比,大肠俞组潜伏期明显缩短(P<0.01),收缩波个数增多(P<0.05),M3和5-HT3A受体阳性表达增强(P<0.05,P<0.01)。结论:电针"天枢""大肠俞"穴均能调整IBS大鼠肠道动力和敏感性,且电针"天枢"穴的调节效应优于"大肠俞",其机制可能与调整结肠内M3受体和5-HT3A受体表达有关。
Objective To observe and compare the effect of electroacupuncture(EA)of"Tianshu"(ST25)and"Dachangshu"(BL25)on intestinal sensitivity and the expression of M3 muscarinic acetylcholine receptor(M_(3R))and5-hydroxytryptamine 3 Areceptor(5-HT_(3A)R)in the colon tissue in irritable bowel syndrome(IBS)rats,so as to explore its mechanism underlying improvement of IBS.Methods Forty newborn Wistar rats were randomly divided into blank control,model,ST25 and BL25 groups(n=10 rats per group).The IBS model was established by joint application of maternal separation,acetic acid enema plus colorectal distension from day 8 to 21 after birth.At the age of 9 weeks,EA(2 Hz/100 Hz,0.1-0.3 mA)was applied to ST25 and BL25 for 20 min,once every other day for 5 times.The modified method was used to record the abdominal withdrawal reflex(AWR),so as to evaluate the visceral sensitivity by referring to the modified Al-Chaer's and colleagues' methods.The threshold(initial wave),the number and peak-peak value of contraction waves(AWR)within 90 swere recorded.The immunoactivity of M_(3R) and 5-HT_(3A)R in the colon tissue was detected by immunohistochemistry.Results After modeling,the latency was obviously shortened,the number and peak-peak values of contraction waves of intestine were significantly increased,and the immunoactivity of M_(3R) and 5-HT_(3A)R was notably up-regulated in the model group compared with the blank control group(P<0.01).After the EA intervention,the latency was obviously prolonged,the peak-peak value in both ST25 and BL25 groups,and the number of contraction waves in the ST25 group were significantly decreased(P<0.01),and the expression levels of M3 and 5-HT_(3A)R were considerably down-regulated in both ST25 and BL25 groups relevant to the model group(P<0.01).The therapeutic effects were significantly better in the ST25 group than in the BL25 group in prolonging the latency and reducing the contraction wave number,and in down-regulating the expression of colonic M_(3R) and 5-HT_(3A)R(P<0.05,P<0.01).Conclusion EA at ST25 and BL25 can reduce the intestinal sensitivity in IBS rats,which is probably associated with its effects in down-regulating the expression of M_(3R) and 5-HT_(3A)R in the colon.The therapeutic effects of ST25 were obviously better than those of BL25 in suppressing intestinal sensitivity and expression of colonic M_(3R) and 5-HT_(3A)R.
Objective To observe and compare the effect of electroacupuncture(EA)of"Tianshu"(ST25)and"Dachangshu"(BL25)on intestinal sensitivity and the expression of M3 muscarinic acetylcholine receptor(M_(3R))and5-hydroxytryptamine 3 Areceptor(5-HT_(3A)R)in the colon tissue in irritable bowel syndrome(IBS)rats,so as to explore its mechanism underlying improvement of IBS.Methods Forty newborn Wistar rats were randomly divided into blank control,model,ST25 and BL25 groups(n=10 rats per group).The IBS model was established by joint application of maternal separation,acetic acid enema plus colorectal distension from day 8 to 21 after birth.At the age of 9 weeks,EA(2 Hz/100 Hz,0.1-0.3 mA)was applied to ST25 and BL25 for 20 min,once every other day for 5 times.The modified method was used to record the abdominal withdrawal reflex(AWR),so as to evaluate the visceral sensitivity by referring to the modified Al-Chaer's and colleagues' methods.The threshold(initial wave),the number and peak-peak value of contraction waves(AWR)within 90 swere recorded.The immunoactivity of M_(3R) and 5-HT_(3A)R in the colon tissue was detected by immunohistochemistry.Results After modeling,the latency was obviously shortened,the number and peak-peak values of contraction waves of intestine were significantly increased,and the immunoactivity of M_(3R) and 5-HT_(3A)R was notably up-regulated in the model group compared with the blank control group(P<0.01).After the EA intervention,the latency was obviously prolonged,the peak-peak value in both ST25 and BL25 groups,and the number of contraction waves in the ST25 group were significantly decreased(P<0.01),and the expression levels of M3 and 5-HT_(3A)R were considerably down-regulated in both ST25 and BL25 groups relevant to the model group(P<0.01).The therapeutic effects were significantly better in the ST25 group than in the BL25 group in prolonging the latency and reducing the contraction wave number,and in down-regulating the expression of colonic M_(3R) and 5-HT_(3A)R(P<0.05,P<0.01).Conclusion EA at ST25 and BL25 can reduce the intestinal sensitivity in IBS rats,which is probably associated with its effects in down-regulating the expression of M_(3R) and 5-HT_(3A)R in the colon.The therapeutic effects of ST25 were obviously better than those of BL25 in suppressing intestinal sensitivity and expression of colonic M_(3R) and 5-HT_(3A)R.
引文
[1]刘美荣,肖瑞飞,彭芝配,等.针刺“足三里”“太冲”穴对腹泻型肠易激综合征大鼠胃肠激素的影响[J].针刺研究,2012,37(5):363-368.
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[6]胡晔.针刺辨证治疗功能性消化不良疗效观察及其对血清胃泌素影响[D].武汉:湖北中医药大学,2012.
[7]裴丽霞,张伟,宋亚芳,等.电针“天枢”穴对感染后肠易激综合征内脏高敏感模型大鼠结肠肥大细胞活化和P物质的影响[J].针刺研究,2018,43(7):419-423.
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[9]林泽斯,王洪琦.肠易激综合征母婴分离模型应用进展[J].中国实验动物学报,2015,23(4):434-439.
[10]闫雪,唐洪梅,张庆业,等.综合因素对肠易激综合征动物模型造模的影响[J].现代医院,2015,15(4):17-19.
[11]李延青,郭玉婷,左秀丽,等.肠易激综合征内脏感觉过敏动物模型的建立[J].胃肠病学和肝病学杂志,2003,12(4):24.
[12]张露芬.实验针灸学[M].北京:化学工业出版社,2010:219-221.
[13]樊江波,畅洪昇,董世芬,等.慢性应激致肠易激综合征大鼠模型的建立与评价[J].中国实验动物学报,2010,18(2):91-95.
[14]沈蕾.精神因素和应激在IBS发病机制中的作用[D].武汉:华中科技大学,2010.
[15]卢璐,袁建业,费晓燕,等.肠易激综合征发病机制研究及治疗进展[J].中国中西医结合消化杂志,2015,23(9):661-665.
[16]王珊,郭孟玮,高誉珊,等.电针“内关”“天枢”穴对肠易激综合征大鼠结肠动力及结肠D 2受体的影响[J].针刺研究,2018,43(1):24-28.
[17]STASI C,BELLINI M,BASSOTTI G,et al.Serotonin receptors and their role in the pathophysiology and therapy of irritable bowel syndrome[J].Tech Coloproctol,2014,18(7):613-621.
[18]贾琼,薛玲.5-羟色胺3受体及其应用的研究进展[J].医学综述,2012,18(10):1444-1447.
[19]HICKS G A,COLDWELL J R,SCHINDLER M,et al.Excitation of rat colonic afferent fibres by 5-HT3receptors[J].J Physiol,2010,544(3):861-869.
[20]GERSHON M D.Nerves,reflexes,and the enteric nervous system:pathogenesis of the irritable bowel syndrome[J].JClin Gastroenterol,2005,39(5Suppl 3):S184.
[21]KOZLOWSKI C M,GREEN A,GRUNDY D,et al.The 5-HT3receptor antagonist alosetron inhibits the colorectal distention induced depressor response and spinal-fos expression in the anaesthetised rat[J].Gut,2000,46(4):474-480.
[22]武蕾蕾,何志鹏.山楂水提物对肠易激综合征大鼠结肠黏膜5-HT和5-HT3R表达的影响[J].牡丹江医学院学报,2011,32(4):6-9.
[23]王斌,程峰涛.M受体亚型和胃肠道平滑肌功能[J].中国病理生理杂志,2001,17(11):1093-1096.
[24]朱文莲,李滢,杨帅,等.同神经节段不同穴位对肠易激综合征效应机制的比较研究[J].中华中医药杂志,2013,28(11):3224-3227.
[2]蔡玮凌.肠易激综合征的中医辨证论治[D].南京:南京中医药大学,2012.
[3]杨明晓,赵凌,杨洁,等.国内针灸病谱研究规律的文献计量学分析及趋势展望[J].针刺研究,2014,39(3):247-251.
[4]吴风华.针刺天枢穴治疗慢传输型便秘疗效分析[J].实用中医药杂志,2012,28(3):175-177.
[5]陆影,杨维华.磁贴天枢穴治疗小儿脾虚泄40例疗效观察[J].中国中西医结合儿科学,2012,4(5):446-448.
[6]胡晔.针刺辨证治疗功能性消化不良疗效观察及其对血清胃泌素影响[D].武汉:湖北中医药大学,2012.
[7]裴丽霞,张伟,宋亚芳,等.电针“天枢”穴对感染后肠易激综合征内脏高敏感模型大鼠结肠肥大细胞活化和P物质的影响[J].针刺研究,2018,43(7):419-423.
[8]徐昔非.电针大肠之俞募穴与俞募合穴治疗功能性腹泻临床疗效的对比观察[D].济南:山东中医药大学,2014.
[9]林泽斯,王洪琦.肠易激综合征母婴分离模型应用进展[J].中国实验动物学报,2015,23(4):434-439.
[10]闫雪,唐洪梅,张庆业,等.综合因素对肠易激综合征动物模型造模的影响[J].现代医院,2015,15(4):17-19.
[11]李延青,郭玉婷,左秀丽,等.肠易激综合征内脏感觉过敏动物模型的建立[J].胃肠病学和肝病学杂志,2003,12(4):24.
[12]张露芬.实验针灸学[M].北京:化学工业出版社,2010:219-221.
[13]樊江波,畅洪昇,董世芬,等.慢性应激致肠易激综合征大鼠模型的建立与评价[J].中国实验动物学报,2010,18(2):91-95.
[14]沈蕾.精神因素和应激在IBS发病机制中的作用[D].武汉:华中科技大学,2010.
[15]卢璐,袁建业,费晓燕,等.肠易激综合征发病机制研究及治疗进展[J].中国中西医结合消化杂志,2015,23(9):661-665.
[16]王珊,郭孟玮,高誉珊,等.电针“内关”“天枢”穴对肠易激综合征大鼠结肠动力及结肠D 2受体的影响[J].针刺研究,2018,43(1):24-28.
[17]STASI C,BELLINI M,BASSOTTI G,et al.Serotonin receptors and their role in the pathophysiology and therapy of irritable bowel syndrome[J].Tech Coloproctol,2014,18(7):613-621.
[18]贾琼,薛玲.5-羟色胺3受体及其应用的研究进展[J].医学综述,2012,18(10):1444-1447.
[19]HICKS G A,COLDWELL J R,SCHINDLER M,et al.Excitation of rat colonic afferent fibres by 5-HT3receptors[J].J Physiol,2010,544(3):861-869.
[20]GERSHON M D.Nerves,reflexes,and the enteric nervous system:pathogenesis of the irritable bowel syndrome[J].JClin Gastroenterol,2005,39(5Suppl 3):S184.
[21]KOZLOWSKI C M,GREEN A,GRUNDY D,et al.The 5-HT3receptor antagonist alosetron inhibits the colorectal distention induced depressor response and spinal-fos expression in the anaesthetised rat[J].Gut,2000,46(4):474-480.
[22]武蕾蕾,何志鹏.山楂水提物对肠易激综合征大鼠结肠黏膜5-HT和5-HT3R表达的影响[J].牡丹江医学院学报,2011,32(4):6-9.
[23]王斌,程峰涛.M受体亚型和胃肠道平滑肌功能[J].中国病理生理杂志,2001,17(11):1093-1096.
[24]朱文莲,李滢,杨帅,等.同神经节段不同穴位对肠易激综合征效应机制的比较研究[J].中华中医药杂志,2013,28(11):3224-3227.