Research Of Metabonomics For The Acupuncture Treatment Of Functional Dyspepsia By He-Mu Acupoint Combination Based On Ultra Performance Liquid Chromatography-Quadrupole-Exactive Orbitrap-Mass Spectrum Technology

OBJECTIVEThis study aims to explore the metabolites changes (Mol.wt.< 1000) in plasma of factional dyspepsia (FD) patients after acupuncture treatment by using UPLC-Q-Exactive Orbitrap-MS technique, therefor to provide bioinformatics evidence for The metabolomics mechanism of He-Mu point prescription in treating FD.METHOD1.Charaterization of plasma metabonomics in FD patients:UPLC-Q-Exactive Orbitrap-MS- based metabonomic technique was used to explore the potential biomarkers in plasma of FD patients, comparing with the health group. MetPA database was used to analyze the metabolic pathways of the above potential biomarkers.2.Metabonomics research of single-acupoint and He-Mu acupoint combination in the treatment of FD:FD patients were randomly allocated to one of the three acupuncture groups (the lower He-sea point (ST36) group, Front-mu acupoint group (CV12) or He-Mu combination group),with a treatment period of 4 weeks. Plasma were collected and analyzed from every subjects at the beginning,2 weeks after treatment and the end of treatment. Metabonomics analysis was used to find out plasma metabolites with statistically significant change before and after treatment in each group, based on UPLC-Q-Exactive Orbitrap-MS metabonomic technique.3.Content analysis of plasma metabolites with statistically significant change before and after treatment among groups:the change of metabolites in each group were analyzed by comparing with those mean content in health group. The trend of the change would contribute to explain the effect of He-Mu point prescription in treating FD.RESULTS1.Compared with health control group, the content of Phenylethylamine and (3R,7R)-1,3,7-Octanetriol decreased obviously in FD patients(P<0.01); the content of Isoamyl nitrite,2-Phenylacetamide, L-Alloisoleucine significantly increased (P>0.01);the content of 2-Hexylglycerol,4-Hydroxy-L-glutamic acid decreased obviously (P<0.05);the content of 1-Pyrroline-2-carboxylic acid, D-Proline significantly increased(P>0.05), The above metabolites are FD related potential biomarkers.The related pathways of FD are phenylalanine metabolism, arginine and proline metabolism.2.In the He-Mu combination group, fifteen biomarkers which had statistically different changes (P<0.05) after two and four weeks of treatment were detected, and ten of these were identified. The level of Isoamyl nitrite、2-Phenylacetamide、 D-Proline、(3R,7R)-1,3,7-Octanetriol、L-Alloisoleucine decreased; the level of 2-Hexylglycerol、4-Hydroxy-L-glutamic acid、12-Ketodeoxycholic acid、 Niacinamide increased.The related pathways of FD treated by He-Mu acupoint combination are phenylalanine metabolism, arginine and proline metabolism.3.In the lower He-sea point group, eleven biomarkers which had statistically different changes (P<0.05) after two and four weeks of treatment were detected, and nine of these were identified. The level of Isoamyl nitrite、2-Phenylacetamide、D-Proline、 (3R,7R)-1,3,7-Octanetriol、L-Alloisoleucine decreased; the level of 2-Hexylglycerol、4-Hydroxy-L-glutamic acid、12-Ketodeoxycholic acid、 Niacinamide increased. The related pathway of FD treated by lower He-sea point is phenylalanine metabolism.4.In the Front-mu acupoint group, eleven biomarkers which had statistically different changes (P<0.05) after two and four weeks of treatment were detected, and ten of these were identified. The level of Isoamyl nitrite、2-Phenylacetamide、 D-Proline、 (3R,7R)-1,3,7-Octanetriol、L-Alloisoleucine decreased; the level of 2-Hexylglycerol、4-Hydroxy-L-glutamic acid、12-Ketodeoxycholic acid、 Niacinamide increased.The related pathways of FD treated by Front-mu acupoint are phenylalanine metabolism, arginine and proline metabolism.CONCLUSION1.For FD patients, many plasma metabolites deviate from the health, among them, Phenylethylamine, (3R,7R)-1,3,7-Octanetriol, Isoamyl nitrite,2-Phenylacetamide, L-Alloisoleucine,2-Hexylglycerol,4-Hydroxy-L- glutamic acid,1-Pyrroline-2-carboxylic acid, D-Proline are FD related potential biomarkers.2.The phenylalanine metabolism, arginine and proline metabolism maybe the potential target metabolic pathways of FD3.The metabolomics mechanism of He-Mu acupoint combination in FD treatment maybe related to regulating the phenylalanine metabolism, arginine and proline metabolism.4.In all groups, acupuncture benignly regulates the plasma metabolites of FD, but He-Mu acupoint combination shows priority in the total amount and concentration.