柴芍六君汤对慢性萎缩性胃炎肝郁脾虚证模型大鼠胃黏膜组织代谢物表达的影响

Effects of Chaishao Liujun Decoction on Metabolite Expression in Gastric Mucosa Tissue of Rats Model of Chronic Atrophic Gastritis with Liver Depression and Spleen Deficiency

目的从代谢组学角度探讨柴芍六君汤改善慢性萎缩性胃炎(CAG)肝郁脾虚证大鼠代谢紊乱的可能作用机制。方法32只SD大鼠随机分为空白组10只、造模组22只。造模组采用化学诱导、饥饱失常和夹尾刺激法建立CAG肝郁脾虚证大鼠模型,造模10周后进行模型评价。将成模大鼠随机分为模型组和柴芍六君汤组,每组10只。柴芍六君汤组予浓度为0.51 g·mL^(-1)柴芍六君汤10 mL·kg^(-1)·天^(-1)灌胃,空白组和模型组予等体积灭菌饮用水灌胃,共给药4周。一般行为学观察,苏木素-伊红(HE)染色观察胃黏膜组织病理形态,核磁共振氢谱技术(^(1)H-NMR)识别胃黏膜组织代谢物表达变化,代谢通路分析。结果造模组大鼠神态疲惫,行为抑郁,大便溏结不调,胃黏膜固有腺体明显萎缩、炎性细胞浸润,符合CAG肝郁脾虚证表现。经柴芍六君汤给药后,模型大鼠一般行为学和胃黏膜组织病理改善。^(1)H-NMR代谢组学检测出15个与CAG肝郁脾虚证密切相关的胃黏膜组织代谢物,柴芍六君汤干预后回调泛酸、2-羟基丁酸、甲基丙二酸、甲硫醇、乳酸、N-甲基-α-氨基异丁酸、溶血磷脂酰胆碱、谷氨酸、N-乙酰基天冬氨酸、顺式5-四氢十六烷基肉碱、腺苷、脂质含量(P>0.05或P>0.01),共得到D-谷氨酰胺和D-谷氨酸代谢,丙氨酸、天冬氨酸和谷氨酸代谢,精氨酸生物合成代谢这3条关键代谢调节通路。结论柴芍六君汤能够保护和修复CAG肝郁脾虚证模型大鼠胃黏膜,回调差异代谢物的紊乱,其主要治疗机制可能与D-谷氨酰胺和D-谷氨酸代谢、丙氨酸、天冬氨酸和谷氨酸代谢、精氨酸生物合成代谢通路调控有关。

Objective To explore the possible mechanism of Chaishao Liujun Decoction in improving the metabolic disorders in rats with chronic atrophic gastritis(CAG)with liver depression and spleen deficiency syndrome from a metabolomic perspective.Methods 32 SD rats were randomly divided into blank group(n=10)and model group(n=22).In the model group,chemical induction,starvation and satiety disorder and tail clamping stimulation were used to establish the rat model of CAG with liver depression and spleen deficiency syndrome.The model rats were randomly divided into model group and Chaishao Liujun Decoction group,with 10 rats in each group.Chaishao Liujun Decoction group was given 0.51 g·mL^(-1)Chaishao Liujun Decoction 10 mL·kg^(-1)·d^(-1)by gavage,and the blank group and model group were given equal volume of sterilized drinking water by gavage for 4 weeks.General behavioral observation,hematoxylin eosin(he)staining was used to observe the pathological morphology of gastric mucosa.^(1)H-NMR was used to identify the changes of metabolites in gastric mucosa,and the metabolic pathway was analyzed.Results The rats in model group showed fatigue,depression,loose stool,obvious atrophy of the intrinsic glands of gastric mucosa and infiltration of inflammatory cells,which were consistent with the syndrome of liver depression and spleen deficiency in CAG.After administration of Chaishao Liujun Decoction,the general behavior and pathological changes of gastric mucosa were improved.Fifteen gastric mucosal metabolites were detected by^(1)H-NMR metabolomics.After intervention,Chaishao Liujun Decoction recalled pantothenic acid,2-Hydroxybutyric acid,methylmalonic acid,methanethiol,LLactic acid,N-methyl-α-aminoisobutyric acid,lysophosphatidylcholine,glutamate,N-acetylaspartate,cis-5-tetrahydrohexadecylcarnitine,adenosine,and lipid content.Three key metabolic regulation pathways were obtained:DGlutamine and D-glutamate metabolism,alanine,aspartate and glutamate metabolism,and arginine biosynthesis(P>0.05 or P>0.01).Conclusion Chaishao Liujun Decoction can protect and repair gastric mucosa of CAG rats with liver depression and spleen deficiency syndrome,and recall the disorder of differential metabolites.Its main therapeutic mechanism may be related to the metabolism of D-Glutamine and D-glutamate metabolism,alanine,aspartate and glutamate metabolism,and arginine biosynthesis pathway.