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

Effects of electropuncture of acupoints on the foot Yangming stomach meridian on metabolic profiling of gastric mucosa in gastric ulcer rats

目的:采用核磁共振氢谱技术(1H; NMR)研究电针胃经穴对应激性胃溃疡大鼠胃黏膜组织代谢物谱的表达。方法:40只SD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,光镜下观察大鼠胃黏膜组织病理形态学的变化,采用1H NMR技术获取胃黏膜组织1H; NMR谱,并利用模式识别方法分析各组间的代谢物谱的差异。结果:与正常组比较,模型组大鼠胃黏膜损伤和胃黏膜组织代谢轮廓差异明显,胃黏膜组织乙酸、肌; 醇、磷酸胆碱、谷氨酸、甘氨酸、乙醇胺、赖氨酸、3-羟基丁酸、缬氨酸、异亮氨酸、牛磺酸、亮氨酸和甘露醇含量上升(P<0.05),乳酸、甲硫氨酸和天; 冬氨酸含量下降(P<0.05);与模型组比较,胃经组和胆经组大鼠胃黏膜得到明显修复,胃经组大鼠胃黏膜组织代谢轮廓更接近正常大鼠,胃黏膜组织甲硫氨; 酸、天冬氨酸含量上升(P<0.05),乙酸、肌醇、磷酸胆碱、谷氨酸、甘氨酸、赖氨酸、异亮氨酸、牛磺酸、乳酸、亮氨酸和甘露醇含量下降(P<0.05; );胆经组大鼠胃黏膜组织3-羟基丁酸、甲硫氨酸含量上升(P<0.05),肌醇、磷酸胆碱、谷氨酸、乙醇胺、异亮氨酸、乳酸、亮氨酸和甘露醇含量下降(; P<0.05)。结论:电针胃经穴可调节胃溃疡大鼠胃黏膜组织特异代谢物的表达,促进胃黏膜损伤的修复。Objective: To research the effects of electropuncture of acupoints on; the foot Yangming stomach meridian on metabolic profiling of gastric; mucosa in rats of gastric ulcer induced by stressusing the 1H nuclear; magnetic resonance (1H NMR). Methods: Forty SD rats were randomly; divided into the control group, the model group, the stomach meridian; group and the gallbladder meridian group. The rat model of gastric ulcer; was induced by the restrained plus cold stress. The pathological changes; in the rat gastric mucosa tissue were observed by light microscope. The; 1H NMR spectroscopy was determined with the 1H NMR technique. And,; meanwhile the differences in the metabolic profiling were identified; with the pattern recognition method. Results: Compared to normal group,; the gastric mucosa damage and gastric mucosa metabolic profiling are; obvious, the concentration of acetic acid, choline, inositol phosphate,; glutamic acid, glycine, ethanolamine, lysine, 3-hydroxy butyric acid,; valine, isoleucine, taurine, leucine and mannitol increase and the; concentration of lactic acid, methionine and aspartic acid decrease in; model group rats (P<0.05). Compared to model group, the gastric mucosa; damage have been recovered in stomach meridian group and gallbladder; meridian group, the metabolic profiling of gastric mocusa tissue in; stomach meridian group are similar to the normal group. The; concentration of methionine, aspartic acid increase and acetic acid,; choline, inositol phosphate, glutamic acid, glycine, lysine, lactic; acid, leucine, isoleucine, taurine, and mannitol decrease in stomach; meridan group (P<0.05). The concentration of 3-hydroxy butyric acid,; methionine increase and choline, inositol phosphate, lactic acid,; leucine, isoleucine, glutamic acid, ethanol amine and mannitol decrease; in gallbladder meridian group (P<0.05). Conclusion: The electropuncture; of acupoints on the foot Yangming stomach meridian could regulate the; metabolic profiling of gastric mucosa, resulting in improving the; gastric mucosa repair of gastric ulcer rats.国家自然科学基金项目; 国家重点基础研究发展计划(973计划)项目; 广东省自然科学基金项目; 深圳市科技计划项

Effects of electro-acupuncture at stomach meridian acupoints on serum and urine metabolic profiles of gastric ulcer model rats

目的:采用核磁共振氢谱技术(1H NMR)研究电针胃经穴对胃溃疡模型大鼠血清和尿液代谢轮廓的影响。方法:40只SD大鼠随机分为空白组、模型组、胃经穴组和胆经穴组,每组10只。除空白组外,利用束缚冷应激法制备大鼠胃溃疡模型,造模成功后用电针分别刺激胃经组和胆经组相关穴位,收集大鼠的血清和尿液,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:与模型组比较,胃经穴组血清中的肌醇、甘氨酸、谷氨酰胺、异亮氨酸、缬氨酸和尿液中的肌酸酐、乳酸、醋酸盐、肌酸、琥珀酸盐、牛磺酸、甘氨酸、苯乙酰甘氨酸、尿囊素、胆碱、三甲胺、甲酸盐、甜菜碱代谢物的表达水平得到了明显地恢复(P<0.05),更接近空白组的代谢水平。结论:电针胃经穴可通过调节胃溃疡大鼠的氨基酸代谢和脂肪代谢促进胃黏膜损伤的修复。Objective: To explore the effects of electro-acupuncture at stomach meridian acupoints on serum and urine metabolic profiles of gastric ulcer model rats by application of 1H nuclear magnetic resonance(1H NMR) spectroscopy. Methods: Forty SD rats were randomly divided into the control group, model group, stomach meridian group and gallbladder meridian group, with 10 rats in each group. The gastric ulcer rat model was established by the restrained cold stress method. After modeling, the model rats were treated with electro-acupuncture at the stomach meridian and gallbladder meridian respectively, the serum and urine of rats were collected to obtain one dimensional 1H NMR spectrum, and the differences of metabolic profiles between these groups were analyzed by the pattern recognition method. Results: Compared to model group, the concentration of myo-inositol, glycine, glutamine, isoleucine, valine in serum from the stomach meridian group have obviously recovered and returning to normal level. The urinary metabolites such as Creatinine, lactic acid, acetic acid salt, acid, succinic acid salt, taurine, glycine, phenylacetyl glycine, allantoin, choline, trimethylamine, formate, betaine have also partly recovered after electro-acupuncture stimulation in the stomach meridian(P<0.05). Conclusion: Electro-acupuncture at stomach meridian acupoints could repair the gastric ulcer lesion by regulating the metabolism of lipid and amino acid in rats with gastric ulcer.国家自然科学基金项目(No.81473751,No.81260556);; 广东省自然科学基金项目(No.2014 A030313005);; 福建省自然科学基金项目(No.2015J05168);; 福建省卫生厅科研项目(No.wzpw201301)~

双蒲散对慢性萎缩性胃炎大鼠胃黏膜细胞TGF-β1/Smad3信号通路的影响

目的 :观察双蒲散对慢性萎缩性胃炎的治疗作用以及胃黏膜细胞TGF/Smad信号通路的影响。方法 :40只SD大鼠随机分为正常组、模型组、双蒲散组和维酶素组,每组10只。采用N-甲基-N`-硝基-N-亚硝基胍(MNNG)方法复制慢性萎缩性胃炎大鼠模型,正常组不处理,模型组只造模不干预,其余两组造模16周后分别灌胃双蒲散和维酶素4周。光镜观察胃黏膜组织病理学,酶联免疫吸附法测定胃黏膜组织Bcl-2、P53、PCNA、Ag-NORs、EGF、TGF-β1与Smad3蛋白的表达。结果 :与正常组比较,模型组大鼠胃黏膜组织Bcl-2、P53、PCNA、Ag-NORs、EGF、TGF-β1的表达皆明显升高,Smad3皆显著下降(P<0.05);与模型组比较,双蒲散组大鼠胃黏膜组织Bcl-2、P53、PCNA、Ag-NORs、EGF、TGF-β1的表达皆明显下降,Smad3皆显著升高(P<0.05),其效应显著高于维酶素组。结论 :双蒲散可通过调节TGF-β1/Smad3信号通路调节慢性萎缩性胃炎大鼠胃黏膜细胞的增殖和凋亡,抑制胃黏膜细胞的异型增生,阻断慢性萎缩性胃炎向胃癌前病变发展。国家自然科学基金项目(81260556);; 江西省自然科学基金项目(20131512040019