基于LC-MS技术的连朴饮干预慢性萎缩性胃炎模型大鼠血清代谢组学分析

LC-MS Based Serum Metabolomics Analysis of Lian Pu Yin Decoction on Intervention in Rats with Chronic Atrophic Gastritis

目的研究连朴饮(Lian Pu Yin Decoction,LPY)对慢性萎缩性胃炎(Chronic atrophic gastritis,CAG)模型大鼠的干预作用,并探索其作用机制。方法采用1-甲基-3-硝基-1-亚硝基胍(MNNG)自由饮用,雷尼替丁合并酒精灌服刺激方法,同时施以饥饱失常等,复制大鼠CAG模型,通过观察大鼠一般状况、胃窦组织形态病理变化评价LPY对模型大鼠的干预作用;采用高效液相色谱串联质谱(High Performance Liquid Chromatography Tandem Mass Spectrometry,HPLC-MS)技术,结合多元统计分析方法,对大鼠血清进行代谢组学分析,探讨LPY干预CAG的作用机制。结果LPY可改善模型大鼠一般状况,胃窦黏膜组织病理学情况。与正常组相比,模型组筛选得到显著差异代谢物有9种,主要使维生素B6代谢、氨基酸代谢发生紊乱。LPY可显著调节模型组差异代谢产物8种:Kynurenic acid,1-Methylnicotinamide、Indole-3-carboxyaldehyde和5′-S-Methylthioadenosine在LPY组中含量显著上调,Guanosine,Indoleacetic acid,2-Hydroxyisocaproic acid和Uracil在LPY组中含量显著下调。特别是上调了色氨酸肠道微生物代谢物Indole-3-carboxyaldehyde调节了机体的炎症免疫反应;LPY可调节烟酸和烟酰胺代谢、氨基酸代谢等,尤其与半胱氨酸和蛋氨酸代谢高度相关。结论LPY对CAG具有治疗作用,其机制可能与调节烟酸盐代谢通路、氨基酸代谢通路有关。其中,半胱氨酸和蛋氨酸代谢可能是连朴饮治疗CAG作用机制的核心通路。

Objective To investigate the effect and the mechanisms of Lian Pu Yin Decoction(LPY)on chronic atrophic gastritis(CAG)model rats.Methods The CAG model of rats was replicated with 1-Methyl-3-nitro-1-nitrosoguanidine(MNNG)by free drinking,ranitidine+alcohol stimulation by gastric perfusion,and random starvation and satiety.The effect of LPY on CAG was evaluated by assessing the general conditions of rats and the morphological and pathological changes of gastric anther.Rat serum was metabolomically analyzed with high performance liquid chromatography tandem mass spectrometry(HPLC-MS)and multivariate statistical analysis to explore the mechanisms.Results LPY decoction improved the general condition and the histopathological condition of gastric antrum in CAG rats.Compared with the normal group,9 major differential metabolites,which mainly disrupted the metabolism of VB6 and amino acid.LPY decoction significantly regulated the content of 8 different metabolites in the model group:Kynurenic acid,1-methylnicotinamide and indole-3-carboxyaldehyde,5′-s-methylthioadenosine,Guanosine,Indoleacetic acid,2-hydroxyisocaproic acid and Uracil.The contents of Kynurenic acid,1-methylnicotinamide and indole-3-carboxyaldehyde,5′-s-methylthioadenosine were significantly elevated,while significant decrease in the contents of Guanosine,Indoleacetic acid,2-hydroxyisocaproic acid and Uracil were observed in the LPY group.Specifically,aldehyde(I3 A)was upregulated for tryptophan intestinal microbial metabolite indole-3-carboxyaldehyde(CMP),which resulted in regulating the inflammatory immune response in the body.LPY regulated nicotinate and nicotinamide metabolism and amino acid metabolism pathways.In particular,cysteine and methionine metabolism are highly related.Conclusion LPY is therapeutically effective on treating CAG.Its mechanism may be related to the regulation of amino acid and nicotinate metabolic pathways.Among them,cysteine and methionine metabolism may be the core pathway of the CAG mechanism of LPY treatment.