基于经脉-脏腑相关的慢性萎缩性胃炎模型大鼠足三里穴区代谢组学特点分析
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摘要
目的:建立并验证慢性萎缩性胃炎(CAG)大鼠模型,运用代谢组学的方法研究CAG大鼠模型代谢组学特征及生物标志物,探讨胃经穴位与胃之间是否存在一定的相对特异性联系。方法:将32只SD大鼠随机分为空白组和模型组,通过MNNG联合乙醇灌胃、饥饱失常法复制CAG大鼠模型。HE染色观察大鼠胃组织病理学变化,采用1H-NMR检测足三里穴区及胃组织代谢物并分析所得数据。结果:造模结束后,模型组大鼠胃黏膜色苍白,黏膜层变薄,弹性减少,皱襞变平,黏液减少,有散在出血点,HE染色结果可见大量炎性细胞浸润,大量主细胞坏死空泡样变化,大量纤维组织充填,并有诸多集中出血点,腺体结构紊乱减少,固有腺体出现萎缩。与空白组对比,模型组大鼠胃组织共有17个差异性代谢物,其中,乳酸、谷氨酰胺、谷胱甘肽、N-乙酰天门冬氨酸、尿苷二磷酸葡萄糖、肌糖、腺嘌呤核糖核苷酸7个代谢物浓度显著升高,而亮氨酸、牛磺酸、N,N-二甲基甘氨酸、甜菜碱、丝氨酸、磷酸胆碱、乙酰磷酸胆碱、尿嘧啶、尿苷、苯丙氨酸10个代谢物浓度显著降低。模型组大鼠足三里穴区组织中谷氨酰胺、琥珀酸、赖氨酸、富马酸、次黄嘌呤、一磷酸腺苷显著降低,而N-乙酰天门冬氨酸、天冬氨酸、天冬酰胺、磷酸胆碱/甘磷酸胆碱、酪氨酸、苯丙氨酸、黄嘌呤、肌苷显著升高。结论:代谢组学得出CAG模型大鼠足三里穴区与胃组织之间在病理上相互关联,可初步表明胃经穴位与胃之间存在一定的相对特异性联系。
Objective: To establish and validate a rat model of chronic atrophic gastritis(CAG), metabolomics is applied to study the metabolomics characteristics and biomarkers of CAG rat model, and to explore whether there is a certain relative specificity relationship between gastric acupoints and stomach. Methods: Thirty-two Sprague-Dawley rats were randomly divided into blank group and model group. The CAG rat model was replicated by MNNG combined with ethanol gavage and abnormal of starvation. The pathological changes of gastric tissue were observed by HE staining. The metabolites of Zusanli(ST36) and stomach tissues were detected by 1 H-NMR and the data were analyzed. Results: After the model was established, the gastric mucosa of the model rats was pale, the mucosa was thinned, the elasticity was reduced, the folds were flattened, the mucus was reduced, these was scattered bleeding point. In HE staining, the pathological examination of the stomach tissue under light microscope showed a large infiltration of inflammatory cells, a large number of primary cells necrotic vacuole-like changes, a large number of fibrous tissue filling, and a number of concentrated bleeding points, glandular structural disorders reduced, the inherent gland atrophy. Compared with the blank group, there were 17 differential metabolites in the gastric tissue of the model group, including lactic acid, glutamine, glutathione, N-acetyl aspartate, uridine diphosphate glucose, muscle sugar, and adenine ribonucleotides, these 7 metabolites were significantly increased. Leucine, taurine, N,N-dimethylglycine, betaine, serine, phosphorylcholine, acetylphosphocholine, uracil, uridine, phenylalanine, the concentration of these 10 metabolites were significantly reduced. Compared with the blank group, the glutamine, succinic acid, lysine, fumaric acid, hypoxanthine and adenosine monophosphate in the Zusanli(ST36) area of the model group were significantly reduced, while N-acetylaspartate, aspartic acid, asparagine, choline phosphate/choline phosphate, tyrosine, phenylalanine, xanthine and inosine were significantly elevated. Conclusion: Metabolomics shows that the CAG model rat Zusanli(ST36) and gastric tissue are pathologically correlated, which may indicate that there is a certain relative specific relationship between gastric acupoints and stomach.
Objective: To establish and validate a rat model of chronic atrophic gastritis(CAG), metabolomics is applied to study the metabolomics characteristics and biomarkers of CAG rat model, and to explore whether there is a certain relative specificity relationship between gastric acupoints and stomach. Methods: Thirty-two Sprague-Dawley rats were randomly divided into blank group and model group. The CAG rat model was replicated by MNNG combined with ethanol gavage and abnormal of starvation. The pathological changes of gastric tissue were observed by HE staining. The metabolites of Zusanli(ST36) and stomach tissues were detected by 1 H-NMR and the data were analyzed. Results: After the model was established, the gastric mucosa of the model rats was pale, the mucosa was thinned, the elasticity was reduced, the folds were flattened, the mucus was reduced, these was scattered bleeding point. In HE staining, the pathological examination of the stomach tissue under light microscope showed a large infiltration of inflammatory cells, a large number of primary cells necrotic vacuole-like changes, a large number of fibrous tissue filling, and a number of concentrated bleeding points, glandular structural disorders reduced, the inherent gland atrophy. Compared with the blank group, there were 17 differential metabolites in the gastric tissue of the model group, including lactic acid, glutamine, glutathione, N-acetyl aspartate, uridine diphosphate glucose, muscle sugar, and adenine ribonucleotides, these 7 metabolites were significantly increased. Leucine, taurine, N,N-dimethylglycine, betaine, serine, phosphorylcholine, acetylphosphocholine, uracil, uridine, phenylalanine, the concentration of these 10 metabolites were significantly reduced. Compared with the blank group, the glutamine, succinic acid, lysine, fumaric acid, hypoxanthine and adenosine monophosphate in the Zusanli(ST36) area of the model group were significantly reduced, while N-acetylaspartate, aspartic acid, asparagine, choline phosphate/choline phosphate, tyrosine, phenylalanine, xanthine and inosine were significantly elevated. Conclusion: Metabolomics shows that the CAG model rat Zusanli(ST36) and gastric tissue are pathologically correlated, which may indicate that there is a certain relative specific relationship between gastric acupoints and stomach.
引文
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[13]吴巧凤,颜贤忠,唐勇,等.代谢组学在我国中医药领域的应用.四川中医,2008,26(3):7-9
[14]李清林,王玲,尹华.基于代谢组学技术的中医药现代研究.中华中医药杂志,2013,28(11):3149-3152
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[17]Takahashi T,Sasaki K,Somfai T,et al.N,N-Dimethylglycine decreases oxidative stress and improves in vitro development of bovine embryos.J Reprod Dev,2016,62(2):209-212
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[20]ChakrabortyG,MekalaP,YahyaD,etal.Intraneuronal N-acetylaspartate supplies acetyl groups for myelin lipid synthesis:Evidence for myelin-associated aspartoacylase.J Neurochem,2001,78(4):736-745
[2]周逸平.经脉-脏腑相关是经络理论的核心.针刺研究,1999,24(3):238-241,197
[3]房静远,刘文忠,李兆申,等.中国慢性胃炎共识意见.中华消化杂志,2007,18(3):674-684
[4]路敏敏,金世禄,刘宝珍,等.慢性萎缩性胃炎的诊断进展.中华全科医学,2015,13(5):823-825,836
[5]佘畅.艾灸与针刺对慢性萎缩性胃炎大鼠胃黏膜保护作用和胃组织代谢组学的对比研究.长沙:湖南中医药大学,2017
[6]钟欢.艾灸与针刺对局部穴区组织免疫及代谢调节机制的对比研究.长沙:湖南中医药大学,2017
[7]徐珊,周嘉鹤,王常松,等.慢性萎缩性胃炎证病结合模型的复制.中国中医药科技,2008,15(1):6-8,5
[8]朱萱萱,史淋峰,吴旭彤,等.实验性慢性萎缩性胃炎脾气虚证模型的建立及不同时期病理形态学的改变.中华中医药学刊,2012,30(2):231-233,449-450
[9]张夏毅,张天生,王海军,等.穴位病理反应与脏腑经络相关研究概况.针刺研究,2007,32(5):355-358
[10]夏建飞,梁琼麟,胡坪,等.代谢组学研究策略与方法的新进展.分析化学,2009,37(1):136-143
[11]陈勤,吴巧凤,方剑乔.代谢组学技术及其在针灸临床疗效机制研究中的进展.中国医学创新,2016,13(4):138-141
[12]王金海,杜小正,方晓丽,等.代谢组学技术在针灸研究中的应用探讨.甘肃中医学院学报,2012,29(1):18-20
[13]吴巧凤,颜贤忠,唐勇,等.代谢组学在我国中医药领域的应用.四川中医,2008,26(3):7-9
[14]李清林,王玲,尹华.基于代谢组学技术的中医药现代研究.中华中医药杂志,2013,28(11):3149-3152
[15]Wiggins T,Kumar S,Markar S R,et al.Tyrosine,phenylalanine,and tryptophan in gastroesophageal malignancy:a systematic review.Cancer Epidemiol Biomarkers Prev,2015,24(1):32-38
[16]Lee M Y,Lin Y R,Tu Y S,et al.Effects of sarcosine and N,Ndimethylglycine on NMDA receptor-mediated excitatory field potentials.J Biomed Sci,2017,24(1):18
[17]Takahashi T,Sasaki K,Somfai T,et al.N,N-Dimethylglycine decreases oxidative stress and improves in vitro development of bovine embryos.J Reprod Dev,2016,62(2):209-212
[18]Moffeti J R,Ross B,Arun P,et al.N-Acetylaspartate in the CNS:From neurodiagnostics to neurobiology.Prog Neurobiol,2007,81(2):89-131
[19]Simone I L,Ruggieri M,Tortelil R,et al.Serum Nacetylaspartate level in amyotrophic lateral sclerosis.Arch Neurol,2011,68(10):1308-1312
[20]ChakrabortyG,MekalaP,YahyaD,etal.Intraneuronal N-acetylaspartate supplies acetyl groups for myelin lipid synthesis:Evidence for myelin-associated aspartoacylase.J Neurochem,2001,78(4):736-745