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

Analysis of the metabolomics characteristics in Zusanli(ST36) area of CAG model rats based on meridian-viscera

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.