Study On The Mechanism Of Effect Of Compatibility Of Magnolia Officinalis And Polygala Tenuifolia On Metabolites In Rats

Objective:To investigate the effects of the aqueous and the alcohol extracts of Mangnolia officinalis,Polygala tenuifolia and their compatibility on the metabolites of urine and feces in rats through metabolomics techniques and multivariate statistical data analysis techniques,and provide a reference to clarify the mechanism of Mangnolia officinalis improving the gastrointestinal motility caused by Polygala tenuifolia.Methods:252 mice were randomly divided into blank control group,solvent control group,Polygala tenuifolia aqueous extract group?1.25,2.50,5.00 g/kg?,Polygala tenuifolia alcohol extract group?1.25,2.50,5.00 g/kg?,Mangnolia officinalis aqueous extract group?2.50,5.00,10.00g/kg?,Mangnolia officinalis alcohol extract group?2.50,5.00,10.00g/kg?,their compatibility with aqueous extract group?3.75,7.50,15.00g/kg?,their compatibility with alcohol extract group?3.75,7.50,15.00g/kg?,to investigate the effects of the aqueous and the alcohol extracts of Mangnolia officinalis,Polygala tenuifolia and their compatibility on gastric emptying rate and intestinal propulsion rate in mice,which would determine the dose of metabolomics experimental drugs.SD rats were randomly divided into blank control group,solvent control group,Polygala tenuifolia aqueous extract group 1.75g/kg,Polygala tenuifolia alcohol extract group 1.75g/kg,Mangnolia officinalis aqueous extract group 3.50g/kg,Mangnolia officinalis alcohol extract group 3.50g/kg,their compatibility with aqueous extract group 5.25g/kg,their compatibility with alcohol extract group 5.25g/kg?based on body surface area?,continuous intragastric administration for 3d,urine and feces within 24 hours after the last administration of each rat were collected.Urine and fecal sample of each rat were detected by ultra-high performance liquid chromatography quadrupole-time of flight-mass spectrometry?UPLC-Q-TOF-MS?.The data of urine and fecal test data in each group were analyzed and the characteristic biomarkers were screened by Progenesis QI V2.0,SIMCA-P 14.0,etc.The online metabolic analysis software Metaboanalyst 4.0 was used to enrich and analyze the related metabolic pathways.Results:?1?The effect of 2:1 ratio of different extracts of Magnolia officinalis and Polygala tenuifolia on gastrointestinal motility in mice,the results showed that,compared with the blank control group,the solvent 5%Tween-80 had no significant effect on gastrointestinal motility in mice?P>0.05?;the domperidone group significantly promoted gastric emptying and intestinal propulsion in mice?P<0.05,P<0.01?;Polygala tenuifolia aqueous extract,alcohol extract with the dose of1.25,2.50,5.00g/kg could all significantly reduce the gastric emptying rate in mice?P<0.01?,Polygala tenuifolia aqueous extract with the dose of 5.00g/kg and Polygala tenuifolia alcohol extract with the dose of 1.25,2.50,5.00g/kg could significantly inhibit the intestinal propulsion?P<0.05,P<0.01?;Mangnolia officinalis aqueous extracts with the dose of 2.50,5.00 and 10g/kg could promote gastric emptying and intestinal propulsion in different degrees?P<0.05,P<0.01?;Mangnolia officinalis alcohol extracts with the dose of 2.50g/kg had no significant effect on gastric emptying and intestinal propulsion in mice?P>0.05?,and Mangnolia officinalis alcohol extracts with the dose of 5.00 and 10g/kg could significantly inhibit gastric emptying?P<0.05,P<0.01?.Mangnolia officinalis alcohol extracts with the dose of2.50,5.00g/kg had no significant effect on intestinal propulsion in mice?P>0.05?,Mangnolia officinalis alcohol extracts with the dose of 10.00 g/kg could significantly inhibited intestinal propulsion in mice?P<0.01?.Compared with the Polygala tenuifolia group,each dose group of their compatibility?Mangnolia officinalis:Polygala tenuifolia=2:1?could alleviate the gastrointestinal motility disorder caused by Polygala.?2?The effect of 2:1 ratio of different extracts of Magnolia officinalis and Polygala on urine and fecal metabolites in rats,the results showed that,20 characteristic metabolic markers such as L-fucose were identified in the urine metabolites of the aqueous extract group.Among them,L-fucose,hydroxyphenylacetic acid,sphingomyelin?d18:1/23:0?and phosphatidylinositol were significantly increased in the Polygala tenuifolia group?P<0.05,P<0.01?;3-methoxy-4-Hydroxyphenyl glycol glucuronide,ceramide?N-sphingosine?was significantly decreased?P<0.05,P<0.01?,and their compatibility significantly reversed the change of the above metabolites?P<0.05,P<0.01?.Their metabolites involved 15 metabolic pathways such as pantothenic acid and CoA biosynthesis,tyrosine metabolism,and starch and sucrose metabolism.16 characteristic metabolic markers such as phenylacetaldehyde were identified in fecal metabolites.Among them,in Polygala tenuifolia group,5,6:8,9-Diepoxyergost-22 ene 3,7beta diol,5b-cholestane-3?,7a,12a,24,25-pentanol,3a,7a,12a-trihydroxy-5b-biliary-24-ene,3a,7?-dihydroxyprostase was significantly up-regulated?P<0.05,P<0.01?;phenylacetaldehyde,3,4-dihydroxyphenylethylene glycol,S-acetyl dihydrogen sulfide Octanamide-E,3-methoxy-4-hydroxyphenylacetaldehyde,3a,6a,7b-trihydroxy-5b-cholanoic acid,stearic acid were significantly decreased,and their compatibility significantly reversed the above metabolites?P<0.05,P<0.01?.Its metabolites involved 10 metabolic pathways such as tyrosine metabolism,bile acid biosynthesis,and vitamin B₆ metabolism.16 metabolites such as 2-aminobenzoic acid were identified in the urine metabolite of the alcohol extract group.Among them,2-hydroxy-3-?4-hydroxyphenyl?acrylic acid,3a,7a,12a-trihydroxy-5b-cholestanoic acid,palmitic acid,and octadecanoic acid were significantly increased in the Polygala tenuifolia group?P<0.01?;these metabolites were significantly decreased after compatibility with Mangnolia officinalis?P<0.01?;the metabolites involved 14metabolic pathways such as biosynthesis of unsaturated fatty acids and linoleic acid metabolism.A total of 14 metabolic markers such as 5-formylaminotetrahydrofolate were identified in fecal metabolites.The 3a,7a,12a-trihydroxy-5-cholestanoic acid was significantly increased in the Polygala tenuifolia group?P<0.05?;the metabolite was significantly decreased in the compatibility group?P<0.01?.Its metabolites involved 14 metabolic pathways such as biosynthesis of fatty acids,linoleic acid metabolism,and vitamin B₆ metabolism.In addition,some metabolites were not detected in each group.The RSD of all identified metabolites in 10 QC samples was1.94%¹0.00%,indicating that the instrument had good stability and the data results are reliable.Conclusions:?1?Polygala tenuifolia aqueous extract and alcohol extract could inhibit gastrointestinal motility in varying degrees,and the alcohol extract had more significant inhibition than aqueous extract.The compatibility of Mangnolia officinalis and Polygala tenuifolia with 2:1?3.75,7.50,15.00 g/kg?could significantly alleviate gastrointestinal motility disorders caused by Polygala tenuifolia.,it presents a"killing"compatibility.?2?The compatibility of Mangnolia officinalis and Polygala tenuifolia with 2:1 relieving gastrointestinal motility disorders caused by Polygala tenuifolia may be related to vitamin B₆ metabolism,tyrosine metabolism,tryptophan metabolism,primary bile acid biosynthesis and energy metabolism.?3?The common regulation mechanism of the compatibility of Mangnolia officinalis,Polygala tenuifolia on urine and fecal metabolism is related to vitamin B₆ metabolism,purine metabolism,tyrosine metabolism,phenylalanine metabolism,and primary bile acid biosynthesis;?4?The differences in the gastrointestinal motility between Mangnolia officinalis,Polygala tenuifolia and their compatibility in aqueous extracts and alcohol extracts may be related to the metabolic pathways such as pantothenate and CoA biosynthesis and sphingolipid metabolism,vitamin B₆ metabolism,primary bile acid biosynthesis.