The Effects Of Electroacupuncture On Serum Metabolism In Mice With Stress-related Gastric Mucosal Lesions Based On Widely Targeted Metabolomics

Objective To compare the repair effect of electroacupuncture(EA)at Zusanli(ST36)with that of EA at Shousanli(LI10)on Stress-Related Gastric Mucosal Lesion(SGML)in mice,and excavate potential mechanism of EA at ST36 and LI10 on the repair of SGML.Methods26 SPF-grade Kunming mice were randomly divided into the control group(Group C),the model group(Group SU),the EA at ST36 group(Group ST36)and the EA at LI10group(Group LI10),with 7 mice in each of Group C and Group SU,and 6 mice in each of Group ST36 and Group LI10.First day,All mice except for Group C were established SGML models by the Restraint Water-Immersion Stress(RWIS)method.At the end of modelling,one mouse was randomly selected from each of the C and SU groups and executed for pathological observation of gastric mucosa by HE staining.From the 2nd day on,mice in groups ST36 and LI10 were subjected to binding and fixation as well as bilateral EA intervention,for 15 min/time,once/d for 7d,while mice in groups C and SU were simply bound and fixed without EA intervention.All mice’s muscle strength were tested after the intervention,then samples of serum and gastric tissue were collected from the mice.Samples of gastric tissues for light microscopy were prepared by paraffin embedding and HE staining,while samples of gastric tissues for transmission electron microscopy were prepared by Spurr resin embedding.Then the morphology and structure of the mucosal layer of gastric tissues were observed under light microscopy and transmission electron microscopy respectively.Finally,LC-MS metabolomics technique was used to detect the metabolic changes of serum in SGML mice,and SIMCA 14.1 software,Metabo Analyst website,HMDB and KEGG database were used to screen the metabolites and analyse related metabolic pathways to excavate important biological information.Results1.Behavioral observation : Compared with group C,there was no significant change in grip strength of mice’s limb in group SU(P > 0.05).Compared with group SU,the grip strength in group ST36 and group LI10 both decreased significantly(P < 0.01).2.Histomorphological observation under light microscope: Compared with group C,the gastric mucosal epithelial cells of mice in group SU were obviously missing,the arrangement of gastric glands was disordered,the tissue was deeply stained and the eosinophilic infiltration was serious.Compared with group SU,the extent of epithelial loss and eosinophil infiltration in the gastric mucosa of mice in both group ST36 and LI10 were reduced,while the gastric glands of mice in group ST36 were more neatly arranged,and the gastric glands of mice in group LI10 were significantly dilated,with an increase in the intercellular space and abnormal hyperplasia in the muscle layer.3.Histomorphological observation under transmission electron microscope: compared with group C,the gastric mucosal epithelial cells in group SU were broken,with cytoplasmic outflow and cells’ loose arrangement;compared with group SU,the degree of damage of gastric mucosal epithelial cells in ST36 and LI10 groups were both reduced,but the cells in group ST36 were arranged more neatly,while the arrangement of cells in group LI10 were disordered and the gap of cells was enlarged.4.Extensive targeted metabolomic assay base on LC-MS : the RWIS method induced imbalance of metabolic concentration of L-histidine,2-guanidino-succinic acid,1-methyl-L-histidine,Trimethylamine,Ornithine and α-ketoglutarate in serum and disturbance of two metabolic pathways,Histidine metabolism and Arginine synthesis,in mice with SGML.EA at the ST36 acupoint could regulate the imbalance of L-histidine,1-methyl-L-histidine and Histidine synthesis in serum of mice with SGML.EA at the LI10 acupoint could regulate the imbalance of Arginine synthesis.ConclusionsThe Restraint Water-Immersion Stress(RWIS)method induced Stress-Related Gastric Mucosal Lesion(SGML)and disturbance of energy metabolism and neurotransmitter-related metabolism in the serum of mice.The repair effect of EA at ST36 on mice of SGML was better than that of EA at LI10,showing effective restoration capabilities on the integrity of mucus layer and epithelial layer on the surface of gastric mucosa,tightness and orderliness of glandular and cellular arrangement,which may be related with its regulation of metabolic disorders of L-histidine and histidine metabolism caused by SGML.The repair effect of EA at LI10 on SGML was mainly reflected in the integrity of the surface mucus layer and epithelial layer,and its repair effect may be related to the regulation of metabolic disorders in the arginine synthesis pathway.