【作者】 杜冬青；

【导师】 高树中；

【作者基本信息】 山东中医药大学 ， 针灸推拿学， 2013， 博士

【摘要】 目的：本研究在前期已证实隔药灸脐治疗原发性痛经有效的基础上，通过进一步临床试验探讨神阙穴与相同神经节段非任脉穴位相比治疗原发性痛经是否具有特异性、药物在隔药灸脐治疗过程中的作用和地位。同时运用LC/MS联用的代谢组学技术检测患者治疗前后血浆和尿液代谢产物的变化，通过比较与对照组代谢产物和途径的差异，初步探明隔药灸脐特异性治疗原发性痛经的效应机制、药物在隔药灸脐治疗原发性痛经中发挥作用的生物学机制。方法：1.将90例原发性痛经患者随机分为隔药灸脐组、隔药灸大横组、艾条灸脐组各30例，采用视觉模拟评分法Visual Analogue Scale(VAS)评定痛经平均腹痛程度，通过COX痛经症状量表（the Cox Menstrual Symptom Scale）进行痛经临床症状评分，比较三组的疗效差异。2.分别从隔药灸脐组、隔药灸大横组、艾条灸脐组中各随机选取10例患者，另选取正常受试者10例作为对照，运用LC/MS联用的代谢组学技术检测患者治疗前后以及正常受试者血浆和尿液的代谢产物，获得隔药灸脐治疗原发性痛经的相关代谢物质，推导其相关代谢通路，并通过比较与对照组代谢产物和途径的差异，建立临床症状、特征代谢产物和途径与神阙穴效应特异性的关系。结果：1.临床试验：一疗程后，三组均能显著降低VAS读数（P<0.01），隔药灸脐组和艾条灸脐组均能显著降低下腹部疼痛总频率（持续总时间）和严重程度评分（P<0.01）；三组下腹部疼痛总频率（持续总时间）、下腹部疼痛严重程度比较显示，隔药灸脐组、艾条灸脐组与隔药灸大横组均有极其显著性差异（P<0.01），隔药灸脐组与艾条灸脐组比较有极其显著性差异（P<0.01）。2.代谢组学研究：三组治疗前后代谢产物均有明显改变。隔药灸脐治疗后雌酮、16-oxestrone、20a-dihydroprogesterone、15-酮-前列腺素F2α、methylimidazoleacetic acid、L-苯丙氨酸、L-酪氨酸、多巴胺-4-硫酸盐、γ-氨基丁酸的含量发生了变化；隔药灸大横组治疗后5-羟基-L-色氨酸、N-acetylvanilalanine、3,4-dihydroxyphenylacetaldehyde、L-酪氨酸、3,4–二羟基苯乙酸的含量发生了变化；艾条灸脐组治疗后脱氢表雄酮硫酸盐、15-keto-13,14-dihydroprostaglandin A2、5’-磷酸吡哆醛、多巴胺-4-硫酸盐、3,4-二羟基苯甲酸、高香草酸、3,4-dihydroxybenzylamine的含量发生了变化。结论：1.神阙穴对胞宫的效应具有特异性，其特异性体现为循经性和相对性；药物是影响神阙穴效应的因素之一。2.对类固醇激素代谢相关的雌酮、16-oxestrone、20a-dihydroprogesterone、花生四烯酸代谢相关的15-酮-前列腺素F2α、免疫相关物质组胺、中枢神经递质γ-氨基丁酸的调节可能是神阙穴特异性调控胞宫的机理所在；隔药灸脐通过对生殖内分泌激素、局部致痛因子、免疫系统相关物质、神经递质的调节，影响机体神经、内分泌、免疫网络从而发挥对痛经的治疗作用，体现了神阙穴的特异性具有系统性的特点。3.应用药物增强了灸脐对生殖内分泌激素中的孕激素、免疫相关物质组胺、中枢神经递质γ-氨基丁酸的调节，可能是药物在隔药灸脐治疗原发性痛经中的作用机制。更多还原

【Abstract】 Objective:Based on the previous studies of herbs-partitioned moxibustion on RN8for primarydysmenorrhea (PD), further studies were carried out. We focused on specificity of acupointeffect of RN8comparing with other non-RN acupoints in the same nerve segment fortreating primary dysmenorrhea, and we also focused on the functions of herbs during thetreatment. Differences of metabolites in plasma and urine of PD patients were compared byusing LC/MS metabonomics technology as to try to explore the specific effect mechanismof herbs-partitioned moxibustion on RN8treatment for PD and the biological mechanismof herbs during the treatment.Methods:a.90cases of PD patients were randomly divided into three groups (herbs-partitionedmoxibustion on RN8group, herbs-partitioned moxibustion on SP15group and moxa stickmoxibustion on RN8group), with30cases in each group. Visual Analogue Scale (VAS)and the Cox Menstrual Symptom Scale (CMSS) were used to evaluate the treatment effectof three groups.b. Plasma and urine samples of30cases (10cases randomly chosen from each group)were tested by using LC/MS metabonomics technology, comparing with samples of10normal persons as control group. Related metabolites were obtained, and the metabolicpathways were deduced. Relationships among clinical symptoms, specific metabolites,metabolic pathways, and specific effect of acupoint RN8were built by comparing thedifferences of metabolites and metabolic pathways.Results:a. Clinical trials: After treatment, scores of VAS were reduced significantly in all threegroups (P<0.01), and scores of total frequency and average severity of lower abdominal pain in the CMSS were reduced significantly in both herbs-partitioned moxibustion on RN8group and moxa stick moxibustion on RN8group (P<0.01). Scores of herbs-partitionedmoxibustion on RN8group and moxa stick moxibustion on RN8group were significantlydifferent from herbs-partitioned moxibustion on SP15group in the CMSS (P<0.01).Scores of herbs-partitioned moxibustion on RN8group were significantly different frommoxa stick moxibustion on RN8group in the CMSS (P<0.01).b. Metabonomics tests: After treatment, metabolites in all three groups variedsignificantly from base lines. Metabolites related to estrone,16-oxestrone,20a-Dihydroprogesterone,15-keto-prostaglandin F2α, methylimidazoleacetic acid,L-phenylalanine L-tyrosine, dopamine-4-sulfate and gamma-aminobutyric acid changed inherbs-partitioned moxibustion on RN8group. Metabolites related to5-hydroxy-L-tryptophan, N-acetylvanilalanine,3,4-dihydroxyphenylacetaldehyde,L-tyrosine and3,4-dihydroxybenzeneacetic acid changed in herbs-partitioned moxibustionon SP15group. Metabolites related to DHEA sulfate,15-keto-13,14-dihydroprostaglandinA2, pyridoxal5’-phosphate, dopamine-4-sulfate,3,4-dihydroxymandelic acid, homovanillicacid,3,4-dihydroxybenzylamine changed in moxa stick moxibustion on RN8group.Conclusion:a. It manifests that the meridian course specificity and relativity of acupoint RN8. Italso induces that the herbs are one of factors affecting the effect of RN8.b. Estrone,16-oxestrone and20a-dihydroprogesterone associated with arachidonicacid metabolism,15-keto-prostaglandin F2αassociated with steroid hormone metabolism,histamine, and gamma-aminobutyric acid may be the mechanism of regulating the functionof the uterus specifically. Herbs-partitioned moxibustion on RN8group affects thenetwork of nervous system, endocrine system and immune system by regulating theendocrine hormones, local algogenic factors and substances associated with immunesystem. These mentioned above may be the biological mechanism of RN8for specificallyregulating the uterus function, and may also reflect the systematic specificity of RN8.c. Herbs applied during the treatment can strengthen the change of progesterone,histamine, and gamma-aminobutyric acid. This may due to the mechanism effect of herbs.更多还原