Chromatography Mass Spectrometry-based Metabolomics Approach To Study Phlegm-dampness Constitution Of Traditional Chinese Medicine

Objective In the present study,we use high performance liquid chromatography coupled with Q-TOF/MS-based metabolomics method,in order to find the symbolic serum me tabolites and the changes in Metabolic pathways of Phlegm-Dampness constitution of TCM,combing glucose and lipid metabolism,homeostasis model assessment insulin resis tance,to explore the formation,development and pathological characteristics of the TCM phlegmy-dampness constitution,and pathogenesis of "obesity was prone to phlegm-dam pness constitution".Methods According to diagnostic criteria of obesity and TCM phlegm-dampness consti tution,collecting normal control group peace constitution 8 cases,overweight group non-phlegm-dampness constitution 18 cases and phlegm-dampness constitution 30 cases,obes ity group phlegm-dampness constitution 25 cases.comparing body fat parameter(Body Mass Index-BMI,waist circumference-WC,waist-to-hipratio-WHR),blood lipid(TG,CHOL,HDL-C,LDL-C),fasting plasma glucose(FPG),fasting serum insulin(FINS)and homeostas is model assessment insulin resistance(Homa-IR)between the 4 groups,and making cor relation analysis with TCM phlegmy-dampness scores.Based on HPLC-Q-TOF/MS,using multivariate statistical analysis to find the difference serum metabolites of phlegm-da mpness constitution,to explore internal pathological features of TCM phlegm-dampness constitution and the pathogenesis of related-disease further.Results 1.Along with 4 groups,BMI,WC,WHR increase in turn basically;BMI,WC,WH R in overweight group phlegm-dampness constitution and obesity group phlegm-damp ness constitution was significantly higher than that in normal control group peace cons titution(P<0.01,P<0.05);BMI,WC in overweight group non-phlegm-dampness constitutio n was significantly higher than that in normal control group peace constitution(P<0.01,P<0.05);BMI,WC,WHR in obesity group phlegm-dampness constitution was significant ly higher than that in overweight group non-phlegm-dampness constitution(P<0.01);Phle gm-Dampness intergral had a positive correlation to BMI,WC,WHR(P<0.01).2.Along with 4 groups,TG increase in turn,while HDL-C decrease;TG in overweight group phle gm-dampness constitution and obesity group phlegm-dampness constitution was signifi cantly higher than that in normal control group peace constitution and overweight grou p non-phlegm-dampness constitution(P<0.05,P<0.01);CHOL in obesity phlegm-dampness constitution was significantly higher than that in normal control group peace constitut ion and overweight group non-phlegm-dampness constitution(P<0.05);HDL-C in overwe ight group phlegm-dampness constitution and obesity group phlegm-dampness constitut ion was significantly higher than that in normal control group peace constitution(P<0.01,P<0.05);LDL-C in each group had no difference;Phlegm-Dampness intergral had a po sitive correlation to TG(P<0.05),had a genetic correlation to HDL-C(P<0.01),while had no correlation to CHOL.3.FINS,Homa-IR in obesity group phlegm-dampness consti tution was significantly higher than that in normal control group peace constitution(P<0.01);Phlegm-Dampness intergral had no correlation to FPG,FINS,Homa-IR.4.Accordin g to metabolomics method,we are incline to regard that:scatter plot,PCA(principal co mponet analysis)score chart of Each group serum sample distribution can distinguish obviously(P<0.05).①Compared with normal control group peace constitution,8 differenti ally expressed metabolites in overweight group non-phlegm-dampness constitution can be identified(P<0.05),Alpha-N-Phenylacetyl-L-glutamine,PGF2a diethyl amide,Propionyl-L-carnitine,Propionyl-L-carnitine,CARYOPHYLLENYL ACETATE were up-regulated exp ression,3-(Uracil-1-yl)-L-alanine,LysoPC(18:3(6Z,9Z,12Z))were down-regulated expressio n.②Compared with normal control group peace constitution,9 differentially expressed metabolites in overweight group phlegm-dampness constitution can be identified(P<0.05),CARYOPHYLLENYL ACETATE,acetonide were up-regulated expression,N-gamma-L-Glutamyl-L-Phenylalanine,Decanoyl-L-carnitine,L-Arginine,O-octanoyl-R-carnitine,N-palm itoyl serine,Phytosphingosine,C 17 Sphinganine were down-regulated expression.③Compa red with normal control group peace constitution,9 differentially expressed metabolites in obesity group phlegm-dampness constitution can be identified(P<0.05),Propionyl-L-ca rnitine,Alpha-N-Phenylacetyl-L-Glutamine,PGF2a diethyl amide were up-regulated expres sion,LysoPC(18:3(6Z,9Z,12Z)),PC(O-18:1(9Z)/0:0),N-Methylethanolamine phosphate,Lyso PE(0:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z)),PE(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/0:0),LysoPC(18:3(9Z,12Z,15Z))were down-regulated expression.④Compared with overweight group non-p hlegm-dampness constitution,11 differentially expressed metabolites in overweight group phlegm-dampness constitution can be identified(P<0.05),L-Carnitine,acetonide were u p-regulated expression,Hydroxyprolyl-Tyrosine,N-Oleoyl-L-Serine,PE(20:4(5Z,8Z,11Z,14Z)/0:0),N-palmitoyl serine,PS(O-18:0/0:0),Decanoyl-L-carnitine,LysoPE(20:3(11Z,14Z,17Z)/0:0)were down-regulated expression.⑤Compared with overweight group phlegm-dampnes s constitution,10 differentially expressed metabolites in obesity group phlegm-dampness constitution can be identified(P<0.05),N-Oleoyl-L-Serine,Alpha-N-Phenylacetyl-L-Glut amine,were up-regulated expression,CARYOPHYLLENYL ACETATE,LysoPC(18:3(6Z,9Z,12Z)),acetonide were down-regulated expression.Conclusion ①Phlegm-dampness constitution of obesity has obviously pathological char acteristics in abnormal body fat distribution,Lipid metabolism disorders,high-FINS leve 1 and insulin resistance;Phlegm-Dampness intergral had a positive correlation to BMI(P<0.01),Phlegm-Dampness of TCM is the susceptible Constitution of obesity.②Accordi ng to metabolomics method,we are incline to regard that:scatter plot of Each group se rum sample distribution can distinguish obviously(P<0,05).③Through the metabolomics study of phlegm-dampness constitution of obesity and overweight,it shows:The symb olic serum metabolites of phlegm-dampness constitution of obesity:Alpha-N-Phenylacety 1-L-Glutamine,Propionyl-L-carnitine,PGF2α diethyl amide were up-regulated expression;L ysoPC(18:3(6Z,9Z,12Z)),PC(O-18:1(9Z)/0:0),LysoPC(18:3(9Z,12Z,15Z)),PE(22:6(4Z,7Z,10 Z,13Z,16Z,19Z)/0:0),N-Methylethanolamine phosphate,LysoPE(0:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z))were down-regulated expression(P<0.05).The symbolic serum metaboli tes of phlegm-dampness constitution of overweight:CARYOPHYLLENYL ACETATE,ace tonide were up-regulated expression;N-gamma-L-Glutamyl-L-Phenylalanine,Decanoyl-L-ca nitine,N-palmitoyl serine,C17 Sphinganine,Phytosphingosine,O-octanoyl-R-carnitine,L-Arg inine were down-regulated expression(P<0.05).④Through the metabolomics study of o verweight group non-phlegm-dampness and phlegm-dampness constitution,it shows:acet onide,N-palmitoyl serine,Decanoyl-L-camitine were common symbolic serum metabolites of normal control group peace constitution,non-phlegm-dampness and phlegm-dampnes s constitution of overweight group,acetonide were up-regulated expression in phlegm-da mpness constitution of overweight group,while others were down-regulated expression i n phlegm-dampness constitution of overweight group(P<0.05),it indicates that people of TCM phlegm-dampness constitution indeed exit the difference serum metabolites.