The Clinical Features And Urinary Metabolomics Studies On Patients With Phlegm Dampness Syndrome Of Polycystic Ovary Syndrome

Objective:Polycystic ovary syndrome(PCOS)is a common endocrine disorder of premenopausal women,the etiology and pathogenesis still remain uncertain.The prevalence of PCOS with phlegm-dampness syndrome is high,its characteristics were more severe than those of other syndromes and is prone to IR.The present study was to analyze the clinical features of phlegm-dampness syndrome of PCOS,and detect the urinary metabolic characteristics of PCOS with phlegm-dampness syndrome based on ultra performance liquid chromatography mass spectrometry technology,explored the pathogenesis,pathogical changes and related biomarkers of phlegm-dampness syndrome of PCOS,which might reveal biological basis and provide evidence for clinical diagnosis and treatment of PCOS with phlegm-dampness syndrome.Methods:The study included clinical research and metabolomics research.617 PCOS patients were collected to conduct a retrospective analysis,which were divided into 393 phlegm-dampness syndrome and 224 nonphlegm-dampness syndrome two groups according to diagnostic criteria of phlegm-dampness syndrome.Based on the results of retrospective research,94 PCOS women with phlegm-dampness syndrome were selected as PCOS group of phlegm-dampness syndrome,and PCOS group with phlegm-dampness syndrome was divided into 48 PIR and 46 PNIR subgroups,50 healthy women were recruited as control group to conduct a prospective study.Clinical parameters of all subjects were assessed,fasting serum samples and urine samples were also collected,sex hormones,insulin,blood glucose and lipid concentrations were measured,and ultra performance liquid chromatography mass spectrometry technology was used to detected the urinary metabolites.Results:1.Results of retrospective study:among 617 PCOS patients,the prevalence of PCOS with phlegm-dampness syndrome was 63.7%,the prevalence of PCOS with nonphlegm-dampness syndrome was 36.3%.The prevalence of IR was 70.48%and 29.02%among PCOS patients of phlegm-dampness syndrome and nonphlegm-dampness syndrome respectively.Compared to PCOS group of nonphlegm-dampness syndrome,PCOS group with phlegm-dampness syndrome had significantly higher levels of BMI,WHR,systolic blood pressure(BP),diastolic BP,prevalence of acanthosis nigricans,seborrhea prevalence,IR prevalence,IFG prevalence,T,FBG,FINS,HOMA-IR,TG,TC,LDL,TC/HDL,TG/HDL,LDL/HDL,LAP(P<0.05);Hirsutism prevalence,LH,FSH,LH/FSH,HDL,SHBG level in PCOS group with phlegm-dampness syndrome were significantly lower than those of PCOS group with nonphlegm-dampness syndrome(P<0.05);The two groups had no significant difference in age,F-G score,acne prevalence,DHEAS,DHEAS/TT and AND level(P>0.05).2.Results of prospective study:PCOS group with phlegm-dampness syndrome and control group had no significant difference in age,menarche age,BMI,HC,systolic BP,diastolic BP(P>0.05);A significant increase of WC,WHR,F-G score,prevalence of hirsutism,prevalence of acanthosis nigricans,acne prevalence,seborrhea prevalence,MS prevalence,IGR prevalence in PCOS group with phlegm-dampness syndrome when compared to control group(P<0.05);PIR group had significantly higher WC,HC,seborrhea prevalence,MS prevalence than those of PNIR group with phlegmdampness syndrome(P<0.05).FSH,DHEAS,DHEAS/TT were no significant difference between PCOS group with phlegm-dampness syndrome and control group(P>0.05);A significant increase of LH,LH/FSH,T,AND,FAI levels and a significant decrease of SHBG level in PCOS group with phlegm-dampness syndrome when compared to control group(P<0.05);PIR women had significantly higher FAI and lower SHBG level than those of PNIR patients with phlegm-dampness syndrome(P<0.05).PCOS patients of phlegm-dampness syndrome and healthy women had no significant difference in 2h glucose,TG,HDL,ApoAI,LDL/ApoB(P>0.05);FBG,FINS,2h insulin,HOMA-IR,TC,LDL,ApoB,LAP,TC/HDL,TG/HDL,LDL/HDL,ApoB/ApoAI levels were significantly increased in PCOS group with phlegm-dampness syndrome than those of control group,and its HDL/ApoAI was significantly decreased than that of control group(P<0.05);FBG,FINS,2h insulin,HOMA-IR,LDL,ApoB,LAP,TC/HDL,TG/HDL,LDL/HDL,ApoB/ApoAI in PIR group were significantly increased than those of PNIR group of phlegm-dampness syndrome,HDL,HDL/ApoAI level were significantly lower than those of PNIR group with phlegm-dampness syndrome(P<0.05).3.Results of urinary metabolomics:Based on ultra performance liquid chromatography coupled with mass spectrometry and statistical methods,the 23 differential metabolites were identified and divided into steroids,fatty acids,carboxylic acids,nucleosides,purines,pyrimidines and“others”seven categories,the differential metabolites involved in steroid hormone metabolism,lipid metabolism,pantothenate and CoA biosynthesis,amino acid metabolism,TCA cycle,vitamin B6 metabolism and so on.Compared to control group,17-hydroxyprogesterone,21-deoxycortisol,cortexolone,tetradecenoylcarnitine,octanoylcarnitine,decanoylcarnitine,hexanoylcarnitine,methylsuccinic acid,3-hydroxy-3-methyl-glutaric acid,L-histidine,pantothenic acid,N-acetylputrescine,aconitic acid,2-methylguanosine,1-methylguanosine,N4-acetylcytidine,2’-deoxyinosine,xanthosine,hypoxanthine,cAMP,4-pyridoxic acid,Ile-Phe-Gly were significantly upregulated,3-hydroxypicolinic acid was significantly downregulated in PCOS group with phlegm-dampness syndrome(P<0.05);Tetradecenoylcarnitine,3-hydroxy-3-methyl-glutaric acid,pantothenic acid,4-pyridoxic acid in PIR group were significantly downregulated than those of PNIR group with phlegm-dampness syndrome(P<0.05).Among MetPA signaling pathways,impact value of pantothenate and CoA biosynthesis,histidine metabolism,steroid hormone biosynthesis,vitamin B6 metabolism and TCA cycles metabolism were 0.18,0.14,0.09,0.06,0.06 respectively,the five metabolic pathways were the potential signaling pathway involved in PCOS with phlegm-dampness syndrome.Cortexolone,21-deoxy cortisol,17-hydroxyprogesterone,tetradecenoylcarnitine,L-histidine,xanthosine,N4-acetylcytidine,2-methylguanosine were the potential diagnostic biomarkers of phlegm-dampness syndrome of PCOS,the ROC value was 0.79,0.77,0.75,0.74,0.73,0.71,0,71,0.702 respectively.Conclusion:1.PCOS patients with phlegm-dampness syndrome demonstrated disorder of sex hormone,had severe abnormality of glucose and lipid metabolism,especially IR patients,and had an increasing risk for MS,T2DM and cardiovascular diseases.2.Based on LC-MS metabolomics approach,the changes of metabolites of PCOS patients with phlegm-dampness syndrome revealed disorder of steroid hormone biosynthesis metabolism,pantothnate and CoA biosynthesis,histidine metabolism,vitamin B6 metabolism,accumulation of acylcarnitines,increased nucleosides metabolism and TCA cycle;The abnormalities of steroid hormone biosynthesis,pantothenate and CoA biosynthesis,histidine metabolism,Vitamin B6 metabolism,TCA cycles and accumulation of acylcartines might involve in PCOS of phlegm-dampness syndrome.Urinary cortexolone,21-deoxycortisol,17-hydroxyprogesterone,tetradecenoylcarnitine,L-histidine,xanthosine,N4-acetylcytidine,2-methylguanosine were defined as biomarkers for distinguishing PCOS patients with phlegm-dampness syndrome from healthy controls.Futhermore,these findings suggest that LC-MS metabolic approach was helpful to understand the pathogenesis and provide a promising strategy for PCOS diagnosis,changes of metabolites might be effective indicators of therapeutic intervations for PCOS with phlegm-dampness syndrome in future.