脐疗对寒凝血瘀型痛经大鼠扭体反应及神阙穴区TRPV1的影响
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摘要
目的观察脐疗对寒凝血瘀型痛经大鼠扭体反应及神阙穴区TRPV1表达的影响,初步探索脐疗效应与神阙穴区的关系及启动机制。方法将50只雌性大鼠随机分为正常对照组、模型组、药物贴脐组、隔药灸脐组、艾条灸脐组,每组各10只。除正常对照组外,其余各组均构建寒凝血瘀型痛经模型,并给予相应的治疗。记录各组大鼠扭体反应情况,通过免疫荧光法、实时荧光定量PCR技术检测各组大鼠神阙穴区TRPV1的表达。结果与模型组比较,脐疗组的扭体潜伏期明显延长(P <0. 01),扭体次数明显减少(P <0. 01),扭体评分降低(P <0. 05或P <0. 01),隔药灸脐组在延长扭体潜伏期、减少扭体次数及降低扭体评分方面优于药物贴脐组与艾条灸脐组(P <0. 05或P <0. 01);模型组TRPV1mRNA表达较正常对照组明显升高(P <0. 01);治疗后隔药灸脐组与艾条灸脐组TRPV1mRNA表达较模型组均明显升高(P <0. 01);对TRPV1mRNA表达的调节作用,艾条灸脐组与隔药灸脐组优于药物贴脐组(P <0. 01)。结论 3种脐疗法均能缓解寒凝血瘀型痛经大鼠的扭体反应,隔药灸脐法作用最优;脐疗通过上调寒凝血瘀型痛经大鼠神阙穴区TRPV1mRNA的表达以激活腧穴功能,诱导神阙信息启动;隔药灸脐法与艾条灸脐法对神阙的激活作用优于药物贴脐法。
Objective To observe the effect of navel therapy on the expression of TRPV1 in Shenque of coagulated cold and blood stasis type dysmenorrhea model rats,to explore the initiation mechanism and the relationship between the umbilical effect and Shenque. Methods Fifty female rats were randomly divided into five groups: group A(normal control group),group B(model group),group C(drug sticking navel group),group D(Herb-partitioned moxibustion on navel group),group E(moxa moxibustion navel group),10 in each group. In addition to group A,coagulated cold and blood stasis dysmenorrhea model was constructed. We gave esch group the corresponding treatment,and the writhing response of rats in each group was recorded. The expression of TRPV1 in Shenque was detected by immunofluorescence and quantitative real-time PCR. Results Compared with group B,the latency of writhing of the umbilical treatment group was significantly prolonged(P < 0. 01),the number of writhing was significantly reduced(P < 0. 01),and the writhing score was decreased(P < 0. 05 or P < 0. 01),group D was superior to group C and group E in prolonged writhing latency,reduction of writhing frequency and reduction of writhing score(P < 0. 05 or P <0. 01). The expression of TRPV1 mRNA in group D and group E were significantly higher than that in group B(P < 0. 01). For the regulation of TRPV1 mRNA expression,group D and group E were superior to group C(P < 0. 01). Conclusion The three kinds of umbilical therapy all can relieve the writhing response of coagulated cold and blood stasis dysmenorrhea rats,herb-partitioned moxibustion on navel therapy has the best effect. Umbilical therapy activates the acupoint function by up-regulating the expression of TRPV1 mRNA in the Shenque of coagulated cold and blood stasis type dysmenorrhoea rat,inducing Shenque information start,the activation of Herb-partitioned moxibustion on navel therapy and and moxa moxibustion navel therapy to Shenque were better than drug sticking navel therapy.
Objective To observe the effect of navel therapy on the expression of TRPV1 in Shenque of coagulated cold and blood stasis type dysmenorrhea model rats,to explore the initiation mechanism and the relationship between the umbilical effect and Shenque. Methods Fifty female rats were randomly divided into five groups: group A(normal control group),group B(model group),group C(drug sticking navel group),group D(Herb-partitioned moxibustion on navel group),group E(moxa moxibustion navel group),10 in each group. In addition to group A,coagulated cold and blood stasis dysmenorrhea model was constructed. We gave esch group the corresponding treatment,and the writhing response of rats in each group was recorded. The expression of TRPV1 in Shenque was detected by immunofluorescence and quantitative real-time PCR. Results Compared with group B,the latency of writhing of the umbilical treatment group was significantly prolonged(P < 0. 01),the number of writhing was significantly reduced(P < 0. 01),and the writhing score was decreased(P < 0. 05 or P < 0. 01),group D was superior to group C and group E in prolonged writhing latency,reduction of writhing frequency and reduction of writhing score(P < 0. 05 or P <0. 01). The expression of TRPV1 mRNA in group D and group E were significantly higher than that in group B(P < 0. 01). For the regulation of TRPV1 mRNA expression,group D and group E were superior to group C(P < 0. 01). Conclusion The three kinds of umbilical therapy all can relieve the writhing response of coagulated cold and blood stasis dysmenorrhea rats,herb-partitioned moxibustion on navel therapy has the best effect. Umbilical therapy activates the acupoint function by up-regulating the expression of TRPV1 mRNA in the Shenque of coagulated cold and blood stasis type dysmenorrhoea rat,inducing Shenque information start,the activation of Herb-partitioned moxibustion on navel therapy and and moxa moxibustion navel therapy to Shenque were better than drug sticking navel therapy.
引文
[1]马玉侠.隔药灸脐法治疗痛经的临床研究[D].山东中医药大学硕士学位论文,2002:9.
[2]孙玉阳,纪宏宇,陈博,等.原发性痛经的发病机制及中医药治疗的研究进展[J].中国药师,2017,20(1):144.
[3] Sznajder KK,Harlow SP,Burgard SA,et al. Gynecologic pain related to occupational stress among female factory workers in Tianjin,China[J]. Int J Occup Environ Health,2014,20(1):33.
[4] Nasehi M,Sehhatie F,Zamanzadeh V,et al. Comparison of the effectiveness of combination of fennel extract/vitamin E with ibuprofen on the pain intensity in students with primary dysmenorrhea[J]. Iran J Nurs Midwifery Res,2013,18(5):355.
[5] Subasinghe AK,Happo L,Jayasinghe YL,et al. Prevalence and severity of dysmenorrhea,and management options reported by young Australian women[J]. Aust Fam Physician,2016,45(11):829.
[6]王艳英.原发性痛经发病机制及治疗的研究进展[J].中华中医药杂志,2015,30(7):2447.
[7]李姝婧,张晓宁,马凤君,等.脐疗对寒凝血瘀型痛经大鼠神阙穴区肥大细胞的影响[J].时珍国医国药,2017,28(9):2288.
[8]杜冬青.隔药灸脐法治疗原发性痛经的经穴效应特异性研究[D].山东中医药大学博士学位论文,2013:93.
[9]盖晓丽.隔药灸脐与艾条灸脐对原发性痛经患者症状及子宫动脉血流动力学的影响[D].山东中医药大学硕士学位论文,2013:8.
[10]薛哲,刘存志,高树中,等.隔药灸治疗原发性痛经及对患者生殖内分泌的影响[J].中国针灸,2014,34(3):209.
[11]许咏思,嵇波,胡琅琳,等.寒凝血瘀证痛经大鼠模型制备和评价方法述评[J].中医药学报,2013,41(4):25.
[12] Schmauss C,Yaksh TL. In vivo studies on spinal opiate receptor systems mediating antinociception. II. Pharmacological profiles suggestinga differential association of mu,delta and kappa receptors with visceralchemical and cutaneous thermal stimuli in the rat[J]. Pharmacol ExpTher,1984,228(1):1.
[13] Stokes AJ,Shimoda LM,Koblan-Huberson M,et al. A TRPV2-PKA signaling module for transduction of physical stimuli in mast cells[J]. J Exp Med,2004,200(2):137.
[14] Turner H,del Carmen KA,Stokes A. Link between TRPV channels and mast cell function[J]. Springer Berlin Heidelberg,2007,179(179):457.
[15]刘珍洪.基于热敏通道TRPV1、TRPA1探讨吴茱萸汤的散寒止痛机理[D].北京中医药大学硕士学位论文,2017:9.
[16] Vay L Gu C,Mc Naughton PA. The thermo-TRP ion channel family:properties and therapeutic implications[J]. Br J Pharmacol,2012,165(4):787.
[17] Ramsey I S,Dellng M,Clapham D E. An introduction to TRP channels[J]. Annu Rev Physiol,2006,68(68):619.
[18]黄海燕,张世忠. TRPV1和TRPV4通道参与缺血/氧处理心血管保护的研究进展[J].药学研究,2016,35(1):37.
[19] ZHENG J. Molechanism of TRP Channels[J]. ComprPhysiol,2013,3(1):221.
[20] NILIUS B. TRP channels in disease[J]. Biochimica EtBiophysica Acta,2007,1772(8):805.
[21]卢圣锋,尹海燕,唐勇,等.瞬时感受器电位及其与艾灸温热效应机制研究结合的思考[J].针刺研究,2012,37(2):151.
[22]朱琏.新针灸学[M].北京:人民卫生出版社,1954.
[2]孙玉阳,纪宏宇,陈博,等.原发性痛经的发病机制及中医药治疗的研究进展[J].中国药师,2017,20(1):144.
[3] Sznajder KK,Harlow SP,Burgard SA,et al. Gynecologic pain related to occupational stress among female factory workers in Tianjin,China[J]. Int J Occup Environ Health,2014,20(1):33.
[4] Nasehi M,Sehhatie F,Zamanzadeh V,et al. Comparison of the effectiveness of combination of fennel extract/vitamin E with ibuprofen on the pain intensity in students with primary dysmenorrhea[J]. Iran J Nurs Midwifery Res,2013,18(5):355.
[5] Subasinghe AK,Happo L,Jayasinghe YL,et al. Prevalence and severity of dysmenorrhea,and management options reported by young Australian women[J]. Aust Fam Physician,2016,45(11):829.
[6]王艳英.原发性痛经发病机制及治疗的研究进展[J].中华中医药杂志,2015,30(7):2447.
[7]李姝婧,张晓宁,马凤君,等.脐疗对寒凝血瘀型痛经大鼠神阙穴区肥大细胞的影响[J].时珍国医国药,2017,28(9):2288.
[8]杜冬青.隔药灸脐法治疗原发性痛经的经穴效应特异性研究[D].山东中医药大学博士学位论文,2013:93.
[9]盖晓丽.隔药灸脐与艾条灸脐对原发性痛经患者症状及子宫动脉血流动力学的影响[D].山东中医药大学硕士学位论文,2013:8.
[10]薛哲,刘存志,高树中,等.隔药灸治疗原发性痛经及对患者生殖内分泌的影响[J].中国针灸,2014,34(3):209.
[11]许咏思,嵇波,胡琅琳,等.寒凝血瘀证痛经大鼠模型制备和评价方法述评[J].中医药学报,2013,41(4):25.
[12] Schmauss C,Yaksh TL. In vivo studies on spinal opiate receptor systems mediating antinociception. II. Pharmacological profiles suggestinga differential association of mu,delta and kappa receptors with visceralchemical and cutaneous thermal stimuli in the rat[J]. Pharmacol ExpTher,1984,228(1):1.
[13] Stokes AJ,Shimoda LM,Koblan-Huberson M,et al. A TRPV2-PKA signaling module for transduction of physical stimuli in mast cells[J]. J Exp Med,2004,200(2):137.
[14] Turner H,del Carmen KA,Stokes A. Link between TRPV channels and mast cell function[J]. Springer Berlin Heidelberg,2007,179(179):457.
[15]刘珍洪.基于热敏通道TRPV1、TRPA1探讨吴茱萸汤的散寒止痛机理[D].北京中医药大学硕士学位论文,2017:9.
[16] Vay L Gu C,Mc Naughton PA. The thermo-TRP ion channel family:properties and therapeutic implications[J]. Br J Pharmacol,2012,165(4):787.
[17] Ramsey I S,Dellng M,Clapham D E. An introduction to TRP channels[J]. Annu Rev Physiol,2006,68(68):619.
[18]黄海燕,张世忠. TRPV1和TRPV4通道参与缺血/氧处理心血管保护的研究进展[J].药学研究,2016,35(1):37.
[19] ZHENG J. Molechanism of TRP Channels[J]. ComprPhysiol,2013,3(1):221.
[20] NILIUS B. TRP channels in disease[J]. Biochimica EtBiophysica Acta,2007,1772(8):805.
[21]卢圣锋,尹海燕,唐勇,等.瞬时感受器电位及其与艾灸温热效应机制研究结合的思考[J].针刺研究,2012,37(2):151.
[22]朱琏.新针灸学[M].北京:人民卫生出版社,1954.