Metabolic Response of Basal Ganglia in Copper-laden Rat to Gandouling Based on ~1H NMR Spectroscopy

采用核磁共振氢谱(1H NMR)技术与多元统计分析相结合的方法,对肝豆灵对大鼠基底节铜损伤的调节机制进行研究。24只雄性Wistar大鼠随机分为正常组、模型组和肝豆灵组,每组8只,以铜负荷法造模。从造模第7周开始,肝豆灵组大鼠以肝豆灵灌胃。数据分析显示:与正常组比较,模型组的基底节细胞凋亡指数显著增高(p<0.01);基底节组织中的谷氨酰胺、尿苷、苏氨酸含量升高(p<0.05),甘露醇、腺苷、N-乙酰天冬氨酸、谷氨酸、N-乙酰谷氨酸(NAG)、天冬酰胺、乙酸、天冬氨酸、肌醇、三磷酸腺苷(ATP)含量降低(p<0.05);与模型组相比,肝豆灵组的基底节细胞凋亡指数显著降低(p<0.01);基底节组织中的谷氨酰胺、尿苷、乙酸、天冬氨酸含量降低(p<0.05),甘露醇、腺苷、苏氨酸、N-乙酰天冬氨酸、谷氨酸、N-乙酰谷氨酸、天冬酰胺、肌醇、ATP含量升高(p<0.05)。研究结果表明,肝豆灵可影响铜负荷大鼠基底节的代谢,对铜损伤具有一定的修复作用,其机制可能是通过调节氨的解毒和兴奋性氨基酸类神经递质的代谢,干预能量代谢,恢复神经元和神经胶质细胞功能。1H NMR spectroscopy combined with multivariate statistical method was introduced in this paper to explore the regulation effect of Gandouling on basal ganglia injury in copper-laden rats. 24 male Wistar rats were randomly divided into control group,model group and Gandouling group,with8 rats in each group. The rat model was established by copper-laden method. From the 7th week of modeling,Gandouling group was given Gandouling formula via gavage. Compared with the control group,the index of apoptosis cells in basal ganglia of model group increased significantly( p < 0. 01),and glutamine,uridine and threonine in basal ganglia were increased( p < 0. 05) while mannitol,adenosine,N-acetylaspartate, glutamate, N-acetylglutamine asparagine, acetate, aspartate, myoinositol and ATP were decreased( p < 0. 05). Compared with the model group,the index of apoptosis cells in basal ganglia of Gandouling group was significantly decreased( p < 0. 01),and glutamine,uridine,acetate and aspartate were reduced( p < 0. 05) while mannitol, adenosine, threonine,N-acetylaspartate,glutamate,N-acetylglutamine,asparagine,myo-inositol and ATP were elevated( p < 0. 05). The results showed that Gandouling could influence the metabolism of basal ganglia in copper-laden rats,which has a certain therapeutic effect on damage caused by copper. The mechanism may be restoration of the functions of neurons and glial cells,through adjusting the detoxification of ammonia and metabolism of excitatory amino acid neurotransmitter and interfering the energy metab-olism.国家自然科学基金(81202691,81201143,81371639);; 福建省自然科学基金(2015Y0032

Study on the central response mechanism in gastric ulcer rats treated by electroacupuncture using ~1H NMR metabolomics techniques

目的:采用核磁共振氢谱技术(~1H; NMR)研究电针胃经穴治疗应激性胃溃疡大鼠大脑皮质、下丘脑、延髓代谢物谱的表达。方法:40只Wistar大鼠随机分为正常组、模型组、胃经组和胆经; 组。利用束缚冷应激法制备应激性胃溃疡模型大鼠,造模成功后用电针分别刺激胃经组和胆经组相关穴位。收集大鼠大脑皮质、下丘脑、延髓,获取一维~1H; NMR谱,并利用模式识别方法分析各组间的代谢轮廓差异。结果:模型组与正常组、胃经组、胆经组大鼠大脑皮质、下丘脑和延髓代谢物谱均有明显区分,其中正; 常组与模型组分离最明显,胃经组最接近正常组,说明其代谢物谱趋向正常。与模型组比较,胃经组大鼠大脑皮质乳酸、萘乙酸、甘露醇等表达上升(P<0.05; ),而肌酸、腺苷、肌苷等表达下降(P<0.05),下丘脑乳酸、肌醇、谷氨酸酯等表达上升(P<0.05),gamma-氨基丁酸等表达下降(P<0.; 05),延髓乳酸、谷氨酸脂、肌醇、丙氨酸等表达上升(P<0.05),而萘乙酸、肌酸等表达下降(P<0.05)。结论:电针胃经穴可调节胃溃疡大鼠大; 脑皮质、下丘脑、延髓的代谢产物表达,大脑皮质、下丘脑和延髓是电针胃经穴治疗胃溃疡的中枢响应位点。Objective: To study the metabolic profiling of the cerebral cortex,; hypothalamus and medulla oblongata from irritable gastric ulcer rats; treated with electroacupuncture on acupoints of the stomach meridian; with 1H nuclear magnetic resonance (~1H NMR) spectroscopy. Methods:; Forty SD rats are randomly divided into the control group, the model; group, the stomach meridian group and the gallbladder meridian group.; The irritable gastric ulcer rat model was established by water-immersion; and restraint stress method. After modeling, the rats from the stomach; meridian group and the gallbladder meridian group were treated with; electroacupuncture on the stomach meridian and gallbladder meridian; respectively. ~1H NMR spectroscopy was applied to detect the metabolic; profiling of the cerebral cortex, hypothalamus and medulla oblongata,; then analyzed metabolic contour differences among the groups using; pattern recognition method. Results: Obvious distinctions were found; between the metabolites spectrum of the cerebral cortex, hypothalamus; and medulla oblongata from the model group, the normal group, the; stomach meridian group and the gallbladder meridian group. More; significant differences were found between the normal group and the; model group, while the metabolites spectrum of the stomach meridian; group was more distinctly close to that in the normal group, indicating; that metabolites spectrum of the stomach meridian group was likely to; normal. Compared with the model group, the concentration of lactic acid,; acetic acid, mannitol increased while creatine, adenosine and inosine; decreased in cerebral cortex of the stomach meridian group (P<0.05), the; concentration of lactic acid, glutamic acid, inositol and alanine; increased while gamma aminobutyric acid decreased in hypothalamus of the; stomach meridian group (P<0.05), the concentration of lactate,; glutamate, myo-lnositol, alanine increased (P<0.05) and naphthylacetic; acid and creatine decreased in medulla oblongata of the stomach meridian; group (P<0.05). Conclusion: Electroacupuncture on acupoints of the; stomach meridian could regulate specific metabolites in the cerebral; cortex, hypothalamus and medulla oblongata, indicating that the cerebral; cortex, hypothalamus and medulla oblongata as the central response site; in the treatment of gastric ulcer by electroacupuncture on acupoints of; the stomach meridian.国家自然科学基金项目; 广东省自然科学基金项目; 深圳市科技计划项