摘要
[目的]分析补中益气汤对高原应激性胃溃疡小鼠胃黏膜损伤的作用,并基于代谢组学探究补中益气汤预防小鼠高原应激性胃溃疡的机制.[方法]使用低压氧舱构建小鼠高原应激性胃溃疡的模型,60只KM小鼠随机分为模型组、正常组、补中益气汤低剂量组、补中益气汤高剂量组、阳性药物组;通过比较各组小鼠的体质量变化、胃黏膜表观形态及胃黏膜苏木精-伊红染色,分析补中益气汤对高原应激性胃溃疡胃黏膜损伤的影响;通过蛋白免疫印迹法检测各组葡萄糖-6-磷酸脱氢酶(G6PDH)、转录因子YY1蛋白的表达情况,分析补中益气汤对低氧相关的G6PDH和YY1蛋白表达的调控作用;基于液相色谱-质谱联用技术对小鼠胃组织样本进行代谢组学分析,统计分析差异代谢物,进行代谢通路富集分析.[结果]与正常组相比,模型组的胃黏膜表观出现红肿,有大量出血点,出现腺体细胞脱落、细胞坏死、炎细胞浸润等现象,经过补中益气汤的预防,胃溃疡现象有较好的改善;模型组中G6PDH和YY1蛋白表达量增加,而补中益气汤处理后表达量明显减少;小鼠胃组织样本代谢组学检测结果表明差异代谢物参与花生四烯酸代谢、5-羟色胺能突触以及环磷酸腺苷信号等通路.[结论]补中益气汤通过调节5-羟色胺能突触、组氨酸代谢、环磷酸腺苷信号、花生四烯酸代谢等通路以及G6PDH和YY1蛋白的表达缓解高原应激性胃溃疡的发生.
[Objective]High-altitude stress ulcer(HSU)is mainly caused by the unique plateau hypoxic environment,which increases the viscosity of gastric mucosal blood,leads to anaerobic metabolism in the stomach,increases the acidity of gastric juice,and aggravates capillaries damage,eventually resulting in gastric mucosal ulcers.The clinical manifestations of HSU include epigastric pain and discomfort,fullness,pantothenic acid,epigastric burning sensation,nausea,belching,and other symptoms.In traditional Chinese medicine,HSU falls under the categories of"epigastric pain"and"stomach bloating"in traditional Chinese medicine.Buzhong Yiqi decoction,recorded in"PI WEI LUN",is an effective prescription for treating HSU,though its therapeutic mechanism remains unclear.Investigating the mechanism of HSU and developing related preventive drugs are of great significance for the health of people living at high altitudes and for foreign tourists.This study analyzes the effect of Buzhong Yiqi decoction on gastric mucosal damage in mice with HSU,and explores the mechanism by which Buzhong Yiqi decoction prevents plateau stress gastric ulcer in mice,based on metabolomics.[Methods]A hypobaric oxygen chamber was used to construct a mouse model of HSU.60 KM mice were randomly divided into five groups:model group,normal group,Buzhong Yiqi decoction low-dose group,Buzhong Yiqi decoction high-dose group,and positive drug group.The effect of Buzhong Yiqi decoction on gastric mucosal damage caused by plateau stress gastric ulcer was analyzed by comparing body mass changes,apparent morphology of gastric mucosa,and hematoxylin-eosin staining of gastric mucosa of mice in each group.Protein expression of glucose-6-phosphate dehydrogenase(G6PDH)and transcription factor YY1 was detected using Western blotting to verify whether Buzhong Yiqi decoction regulates the expression of G6PDH and YY1 proteins related to hypoxia.Metabolite analysis of mouse gastric tissue samples was performed using liquid chromatography-mass spectrometry technology,with differential metabolites statistically analyzed,and metabolic pathway enrichment analysis conducted.[Results]Pathological analysis showed that compared with the normal group,the gastric mucosa of the model group appeared red and swollen,with a large number of blood stasis spots.Pathological sections showed gland cell detachment,cell necrosis,inflammatory cell infiltration,and other phenomena.In the Buzhong Yiqi low-dose decoction group,the gastric mucosa of mice had a small number of petechiae and slight swelling;in the Buzhong Yiqi high-dose decoction group,the gastric mucosa appeared light pink,with no blood stasis observed.Pathological sections showed that the gastric mucosa of the mice in the Buzhong Yiqi decoction group was relatively intact,with glandular tissue relatively neatly arranged,fewer glands shed,and a reduced number of necrotic cells and inflammatory cells.Western blotting results showed increased expression of G6PDH and YY1 proteins in the model group,while the expressions significantly dereased in the Buzhong Yiqi decoction group.Metabolomics analysis identified a total of 715 metabolites in the normal group,model group,and Buzhong Yiqi decoction group.According to principal component analysis and partial least squares-discriminant analysis,the metabolic profiles of the three groups were different.Further analysis revealed 50 differential metabolites in the Buzhong Yiqi decoction group,which had regulatory effects on the model group.The differential metabolites between each group were further enriched and analyzed using the KEGG Pathway database.The results showed that the differential metabolites were involved in serotonergic synapse,histidine metabolism,cyclic adenosine monophosphate(cAMP)signaling pathway,arachidonic acid metabolism,and adipocyte lipolysis regulation,sphingolipid signaling pathways,and other pathways.[Conclusions]Buzhong Yiqi decoction can promote the repair of gastric mucosal damage and relieve plateau stress by regulating 5-hydroxytryptamine synapses,histidine metabolism,cAMP signaling pathways,arachidonic acid metabolism,and other pathways,as well as by regulating the expression of G6PDH and YY1 proteins to alleviate the occurrence of plateau stress gastric ulcer.Due to the plateau environment and local eating habits,HSU often suffers from complex conditions such as blood stasis and phlegm-dampness.This study focused only on Qi deficiency type and conducted a preliminary study on the mechanism of action of Buzhong Yiqi decoction.In the future,further in-depth research is needed on the detailed mechanism of Buzhong Yiqi decoction s action and its effectiveness in treating different conditions such as blood stasis and phlegm.
作者
刘娜
梁佳林
李积良
王芬祥
奚胜艳
赵成周
王英杰
LIU Na;LIANG Jialin;LI Jiliang;WANG Fenxiang;XI Shengyan;ZHAO Chengzhou;WANG Yingjie(College of Medicine,Qinghai University,Xining 810016,China;College of Ecological and Environmental Engineering,Qinghai University,Xining 810016,China;Department of Anus and Intestines,Qinghai Provincial Hospital of Traditional Chinese Medicine,Xining 810016,China;College of Tibetan Medicine,Qinghai University,Xining 810016,China;School of Medicine,Xiamen University,Xiamen 361102,China)
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2024年第5期943-953,共11页
Journal of Xiamen University：Natural Science
基金
青海省基础研究计划(2023-ZJ-765)。
关键词
代谢组学
高原应激性胃溃疡
补中益气汤
metabolomics
high-altitude stress ulcer
Buzhong Yiqi decoction
