少阳经穴针刺治疗偏头痛的代谢组学研究
摘要
目的：
本研究采用基于核磁共振(NMR)的代谢组学(Metabonomics)方法。分别研究急性偏头痛大鼠模型电针治疗前后的血浆代谢谱图改变和无先兆偏头痛患者针刺治疗前后的血浆代谢谱图改变。通过比较少阳经特定穴与少阳经非穴电针治疗对急性偏头痛大鼠模型血浆~1H-NMR谱图的影响、少阳经特定穴针刺治疗对无先兆偏头痛患者血浆~1H-NMR谱图的影响,探讨少阳经特定穴对偏头痛治疗作用的特异性及其内在生物机制。
方法：
1、50只SD雌性大鼠,随机分为偏头痛模型对照组、循经特定穴组、循经非穴组、正常对照组和生理盐水对照组。按CRISTINA方法皮下注射硝酸甘油(NTG)制作急性偏头痛大鼠模型。循经特定穴组以电针“外关”、“阳陵泉”作为治疗方法；循经非穴组以电针“少阳经非穴”作为治疗方法。运用~1H-NMR技术获取急性偏头痛大鼠血浆~1H-NMR谱图,运用模式识别技术对谱图进行分析,并进行组间比较。
2、纳入无先兆偏头痛女性患者9名,毫针针刺角孙、外关、阳陵泉、丘墟连续治疗5天,运用~1H-NMR技术获取无先兆偏头痛患者针刺治疗前后~1H-NMR谱图,运用模式识别技术对谱图进行分析。比较无先兆偏头痛患者针刺治疗前后血浆~1H-NMR谱图变化,并与健康女性血浆~1H-NMR谱图进行比较。
结果：
1.硝酸甘油注射对大鼠体内的物质代谢有显著影响：与正常组比较,硝酸甘油诱导的偏头痛大鼠模型血浆中Lac(乳酸)、3-HB(β-羟丁酸)、Glucose(葡萄糖)、Glu/Gln(谷氨酸/谷氨酰胺)、Cre(肌酐)等物质含量升高；脂类(Lipid(脂类)、LDL/VLDL(低密度脂蛋白／极低密度脂蛋白))、N-乙酰糖蛋白(Nac)、丙氨酸(Ala)含量下降；
2.生理盐水组大鼠血浆内小分子代谢产物的种类和含量与正常组接近,与模型组存在明显差异；
3.循经特定穴电针能够降低偏头痛大鼠异常升高的Lac、Glucose、Glu/Gln(谷氨酸／谷氨酰胺)含量,使其血浆水平向正常水平靠近；循经非穴电针也能降低偏头痛大鼠异常升高的Lac含量,但对Glucose、Glu/Gln没有调整作用；
4.偏头痛患者与健康人比较,血浆Glu/Gln、Ala(丙氨酸)、UFA(不饱和脂肪酸),Cho(胆碱)、VLDL/LDL含量升高；血浆Lac、Glucose、Nac含量降低；
5.经少阳经特定穴针刺治疗后,偏头痛患者体内的Glu/Gln、Nac、Oac、Ala等物质含量均向对照组靠近,针刺对Lac、Glucose以及脂类物质水平改善不明显。
结论：
1.硝酸甘油皮下注射所引起的血浆小分子物质变化可能与继发于血管扩张、神经源炎症和疼痛等反应。少阳经特定穴电针治疗对急性偏头痛大鼠血浆内小分子物质的改善作用优于少阳经非穴。少阳经特定穴对偏头痛状态下异常升高的血浆谷氨酸、葡萄糖水平具有调整作用,这种作用与少阳经非穴比较显示出特异性。
2.无先兆偏头痛患者体内的高谷氨酸水平可能与无先兆偏头痛患者体内的痛觉敏感和血小板功能异常有关；少阳经特定穴针刺对无先兆偏头痛患者体内的物质代谢具有良性调整作用。偏头痛患者体内的糖类、脂类物质水平是否与偏头痛存在必然联系需要进一步研究证实。
Objective:
Used the nuclear magnetic resonance (NMR) based metabonomic technology to determine the metabolic profiles in the plasma samples of acute migraine rat models and compare the plasma ~1H-NMR metabolic profiles in different acupuncture groups. Used the NMR-based metabonomic technology to determine the metabolic profiles in the migraineurs'plasma samples and observe the changes in plasma ~1H-NMR metabolic profiles after acupuncture therapy. To explore the biologic features of specific points belong Shaoyang Meridians for migraine therapy.
Methods:
1. 50 female SD rats were randomly administrated to five groups:normal control group,sodium injection control group,acute migraine model group, acupuncture group1 (electro-acupuncture therapy with special points belong to Shaoyang Meridian) and acupuncture group 2(electro-acupuncture therapy with none-points along Shaoyang Meridian). The acute migraine model rats were induce by Nitroglycerin (NTG) injection according to the CRISTINA report. Plasma sample was collected after acupuncture treatment and determeined by ~1H-NMR, he metabolic profiles were analyzed by pattern recognition technology.The plasma'H-NMR metabolic profiles were compared between different groups.
2. Including 9 female migraineur without aura, seleced special points belong to Shaoyang Meridian for acupuncture treatments, plasma sample was collected before and after acupuncture treatment and determined by ~1H-NMR, the metabolic profiles were analyzed by pattern recognition technology. The plasma ~1H-NMR metabolic profiles before and after acupuncture treatment had been compared to plasma~1H-NMR metabolic profiles of healthy female.
Result:
1.There are special metabonomics spectras of plasma in acute migraine rats models. The significant different in the plasma ~1H-NMR metabolic profiles resulted in increase in Lac(lactic acid),bata hydroxybutyric acid (3-HB),Glucose,glutamic acid/glutamine (Glu/Gin), creatinine(Cre) level and decrease in lipid, VLDL/LDL, Choline (Cho), N-acetyl-glycoprotein (Nac)
2. The plasma'H-NMR metabolic profiles in sodium injection control group was less abnormal compred to nomal control group, and had obvious difference to NTG injection rats'; 3. The plasma ~1H-NMR metabolic profiles in different acute migraine rats groups indicated that the Lac, Glucose,Glu/Gln content were decreased in acupuncture group 1; the Lac content in in acupuncture group 2 also decreased, but the Glucose,Glu/Gln content had no obvious change in acupuncture group2;
4. The plasma ~1H-NMR metabolic profiles of migraineurs without aura were compared to healthy female'.There were significant decrease in the contents of Lac、Glucose、Nac and increase in Glu/Gln、alanine (Ala)、unsaturated fatty acid (UFA)，Cho、VLDL/LDL in migraineurs'plasma;
5. The acupucnture therapy had effects on adjusting the variation on Glu/Gln、Nac、Oac、Ala in migrainures'plasma to normal control levle, but had few effects on Lac, Glucose and Lipid level.
Conclusion:
1. The NTG injection might cause the blood vessel dilatation and induce the pain and neurogenic inflammation and caused the plasma ~1H-NMR metabolic profiles change.The acupuncture group1 were superior than acupuncture group2 in adjusting the micromolecular substance(Glucose,glutamic acid) content in migraine rats' plasma.Thespecial effects of special points belong to Shaoyang Meridian for migraine might related to the adjusting effects on Glucose,glutamic acid).
2. The abnormal high level of glutamic acid might related to the peripheral algesia sensitization and disfunction of platelet;specific points along Shaoyang Merdian could regulate the variation on micromolecular substance content in migrainures' plasma. The Glucose and Lipid results in migrainures'plasma should be confirmed in the future studies.
本研究采用基于核磁共振(NMR)的代谢组学(Metabonomics)方法。分别研究急性偏头痛大鼠模型电针治疗前后的血浆代谢谱图改变和无先兆偏头痛患者针刺治疗前后的血浆代谢谱图改变。通过比较少阳经特定穴与少阳经非穴电针治疗对急性偏头痛大鼠模型血浆~1H-NMR谱图的影响、少阳经特定穴针刺治疗对无先兆偏头痛患者血浆~1H-NMR谱图的影响,探讨少阳经特定穴对偏头痛治疗作用的特异性及其内在生物机制。
方法：
1、50只SD雌性大鼠,随机分为偏头痛模型对照组、循经特定穴组、循经非穴组、正常对照组和生理盐水对照组。按CRISTINA方法皮下注射硝酸甘油(NTG)制作急性偏头痛大鼠模型。循经特定穴组以电针“外关”、“阳陵泉”作为治疗方法；循经非穴组以电针“少阳经非穴”作为治疗方法。运用~1H-NMR技术获取急性偏头痛大鼠血浆~1H-NMR谱图,运用模式识别技术对谱图进行分析,并进行组间比较。
2、纳入无先兆偏头痛女性患者9名,毫针针刺角孙、外关、阳陵泉、丘墟连续治疗5天,运用~1H-NMR技术获取无先兆偏头痛患者针刺治疗前后~1H-NMR谱图,运用模式识别技术对谱图进行分析。比较无先兆偏头痛患者针刺治疗前后血浆~1H-NMR谱图变化,并与健康女性血浆~1H-NMR谱图进行比较。
结果：
1.硝酸甘油注射对大鼠体内的物质代谢有显著影响：与正常组比较,硝酸甘油诱导的偏头痛大鼠模型血浆中Lac(乳酸)、3-HB(β-羟丁酸)、Glucose(葡萄糖)、Glu/Gln(谷氨酸/谷氨酰胺)、Cre(肌酐)等物质含量升高；脂类(Lipid(脂类)、LDL/VLDL(低密度脂蛋白／极低密度脂蛋白))、N-乙酰糖蛋白(Nac)、丙氨酸(Ala)含量下降；
2.生理盐水组大鼠血浆内小分子代谢产物的种类和含量与正常组接近,与模型组存在明显差异；
3.循经特定穴电针能够降低偏头痛大鼠异常升高的Lac、Glucose、Glu/Gln(谷氨酸／谷氨酰胺)含量,使其血浆水平向正常水平靠近；循经非穴电针也能降低偏头痛大鼠异常升高的Lac含量,但对Glucose、Glu/Gln没有调整作用；
4.偏头痛患者与健康人比较,血浆Glu/Gln、Ala(丙氨酸)、UFA(不饱和脂肪酸),Cho(胆碱)、VLDL/LDL含量升高；血浆Lac、Glucose、Nac含量降低；
5.经少阳经特定穴针刺治疗后,偏头痛患者体内的Glu/Gln、Nac、Oac、Ala等物质含量均向对照组靠近,针刺对Lac、Glucose以及脂类物质水平改善不明显。
结论：
1.硝酸甘油皮下注射所引起的血浆小分子物质变化可能与继发于血管扩张、神经源炎症和疼痛等反应。少阳经特定穴电针治疗对急性偏头痛大鼠血浆内小分子物质的改善作用优于少阳经非穴。少阳经特定穴对偏头痛状态下异常升高的血浆谷氨酸、葡萄糖水平具有调整作用,这种作用与少阳经非穴比较显示出特异性。
2.无先兆偏头痛患者体内的高谷氨酸水平可能与无先兆偏头痛患者体内的痛觉敏感和血小板功能异常有关；少阳经特定穴针刺对无先兆偏头痛患者体内的物质代谢具有良性调整作用。偏头痛患者体内的糖类、脂类物质水平是否与偏头痛存在必然联系需要进一步研究证实。
Objective:
Used the nuclear magnetic resonance (NMR) based metabonomic technology to determine the metabolic profiles in the plasma samples of acute migraine rat models and compare the plasma ~1H-NMR metabolic profiles in different acupuncture groups. Used the NMR-based metabonomic technology to determine the metabolic profiles in the migraineurs'plasma samples and observe the changes in plasma ~1H-NMR metabolic profiles after acupuncture therapy. To explore the biologic features of specific points belong Shaoyang Meridians for migraine therapy.
Methods:
1. 50 female SD rats were randomly administrated to five groups:normal control group,sodium injection control group,acute migraine model group, acupuncture group1 (electro-acupuncture therapy with special points belong to Shaoyang Meridian) and acupuncture group 2(electro-acupuncture therapy with none-points along Shaoyang Meridian). The acute migraine model rats were induce by Nitroglycerin (NTG) injection according to the CRISTINA report. Plasma sample was collected after acupuncture treatment and determeined by ~1H-NMR, he metabolic profiles were analyzed by pattern recognition technology.The plasma'H-NMR metabolic profiles were compared between different groups.
2. Including 9 female migraineur without aura, seleced special points belong to Shaoyang Meridian for acupuncture treatments, plasma sample was collected before and after acupuncture treatment and determined by ~1H-NMR, the metabolic profiles were analyzed by pattern recognition technology. The plasma ~1H-NMR metabolic profiles before and after acupuncture treatment had been compared to plasma~1H-NMR metabolic profiles of healthy female.
Result:
1.There are special metabonomics spectras of plasma in acute migraine rats models. The significant different in the plasma ~1H-NMR metabolic profiles resulted in increase in Lac(lactic acid),bata hydroxybutyric acid (3-HB),Glucose,glutamic acid/glutamine (Glu/Gin), creatinine(Cre) level and decrease in lipid, VLDL/LDL, Choline (Cho), N-acetyl-glycoprotein (Nac)
2. The plasma'H-NMR metabolic profiles in sodium injection control group was less abnormal compred to nomal control group, and had obvious difference to NTG injection rats'; 3. The plasma ~1H-NMR metabolic profiles in different acute migraine rats groups indicated that the Lac, Glucose,Glu/Gln content were decreased in acupuncture group 1; the Lac content in in acupuncture group 2 also decreased, but the Glucose,Glu/Gln content had no obvious change in acupuncture group2;
4. The plasma ~1H-NMR metabolic profiles of migraineurs without aura were compared to healthy female'.There were significant decrease in the contents of Lac、Glucose、Nac and increase in Glu/Gln、alanine (Ala)、unsaturated fatty acid (UFA)，Cho、VLDL/LDL in migraineurs'plasma;
5. The acupucnture therapy had effects on adjusting the variation on Glu/Gln、Nac、Oac、Ala in migrainures'plasma to normal control levle, but had few effects on Lac, Glucose and Lipid level.
Conclusion:
1. The NTG injection might cause the blood vessel dilatation and induce the pain and neurogenic inflammation and caused the plasma ~1H-NMR metabolic profiles change.The acupuncture group1 were superior than acupuncture group2 in adjusting the micromolecular substance(Glucose,glutamic acid) content in migraine rats' plasma.Thespecial effects of special points belong to Shaoyang Meridian for migraine might related to the adjusting effects on Glucose,glutamic acid).
2. The abnormal high level of glutamic acid might related to the peripheral algesia sensitization and disfunction of platelet;specific points along Shaoyang Merdian could regulate the variation on micromolecular substance content in migrainures' plasma. The Glucose and Lipid results in migrainures'plasma should be confirmed in the future studies.
引文
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