The Efficacy And Mechanism Of Qilingwenshenxiaonang Decoction On Patients With Phlegm-dampness Syndrome Of Polysystic Ovary Syndrome Based On Metabonomics

Objective:1.Search the serum differential metabolites between PCOS patients with phlegm-dampness syndrome and healthy women,and the potential biomarkers of PCOS patients with phlegm-dampness syndrome by metabonomics technology,analyze the relevant metabolic pathways,to provide basis for its clinical diagnosis and treatment.2.Treating PCOS patients with phlegm-dampness syndrome with Qilingwenshenxiaonang decoction for six months,observe the clinical efficacy and changes in serum metabolites,analyze the relevant metabolic pathways and explore the possible mechanism of Qilingwenshenxiaonang decoction,to provide a methodological exploration for the modernization of TCM compound prescriptions.Methods:80 cases of PCOS patients with phlegm-dampness syndrome and 40 cases of BMI matched healthy women were included in this study.Collect their serum samples and detect serum differential metabolites between the two groups based on UPLC-MS.The 80 cases of PCOS patients with phlegm-dampness syndrome were randomly divided into treatment group and placebo group with 40 cases each group.This research only studied the clinical efficacy and changes of serum metabolites in the treatment group before and after three months and six months of treatment.Patients in the treatment group were treated with Qilingwenshenxiaonang decoction for six months.Observe the change of TCM syndrome scores,anthropometrics,sex hormones,glucose and lipid metabolism indexes in the treatment group before and after treatment,and evaluate clinical efficacy and safety.At the same time,collect the serum of patients in the treatment group before and after three and six months of treatment for UPLC-MS metabolomic analysis.Results:1.Serum metabonomics results of PCOS patients with phlegm-dampness syndrome:① Differential metabolites:A total of 63 differential metabolites were identified between the PCOS phlegm-dampness syndrome group and the healthy control group.The differential metabolites were involved in androgen metabolism,fatty acid metabolism,fatty amide metabolism,carnitine metabolism,phospholipid metabolism,amino acid metabolism and so on.Compared with the healthy control group,serum 5α-dihydrotestosterone sulfate,FFA,fatty acid amide,carnitine,PC 30:0,PC 32:1,PC 32:2,PC 34:4,PC 36:5,SM 36:1,glutamic acid,KYN,phenylalanine proline,lactic acid and uric acid were significantly upregulated(P<0.05);serum PC 33:2,PC 40:7,LPC,LPE,arginine,glutamine,hippuric acid,phenylacetylglutamine,sulfuric acid,p-cresol,GDCA,GUDCA and TDCA were significantly downregulated(P<0.05).②MetPA signal pathways:Potential signal pathways involved in PCOS with phlegm-dampness syndrome include alanine,aspartate and glutamate metabolism,arachidonic acid metabolism,arginine and proline metabolism,D-glutamine and D-glutamate metabolism,pyruvate metabolism,glycerophospholipid metabolism,and aminoacyl-tRNA biosynthesis.③Potential biomarkers:Uric acid,palmitoyl carnitine,nervonic acid,octadecenoyl carnitine,palmitic acid,glutamic acid,PC.32:1,eicosadienoic acid,eicosatrienoic acid and phenylalanineproline were the potential biomarkers of PCOS with phlegm-dampness syndrome.2.Clinical research results of Qilingwenshenxiaonang decoction intervene in PCOS patients with phlegm-dampness syndrome:Of the 40 patients in the treatment group,37 patients completed three months of treatment and 35 patients completed six months of treatment.①Clinical characteristics:After three months and six months of treatment the mF-G,acanthosis and acne scores,and seborrhea prevalence were significantly lower than before treatment,and the improvement of mF-G score after six months of treatment was better than after three months of treatment(P<0.05).②TCM syndrome scores:After three months and six months of treatment,the total scores of patients decreased significantly,and the efficacy after six months of treatment was better than after three months of treatment(P<0.05).③Sex hormones levels:After three months of treatment,the levels of SHBG and FAI of the patients were significantly improved than before treatment.After six months of treatment,the levels of AND and DHEAS were significantly lower than before treatment,and the effect is better than after three months of treatment.After six months of treatment,the levels of LH of the patients was significantly lower than after three months of treatment(P<0.05).④ The levels of glucose and lipid metabolism:After six months of treatment,the level of GLU2h was significantly higher than before treatment.After three and six months of treatment,the levels of TG and LDL were significantly lower than before treatment(P<0.05).3.Serum metabonomics results of Qilingwenshenxiaonang decoction intervene in PCOS patients with phlegm-dampness syndrome:① Differential metabolites:In the treatment group,serum palmitic acid,oleic acid,eicosadienoic acid,docosadienoic acid,carnitine and Cr were significantly downregulated after three months of treatment with Qilingwenshenxiaonang decoction(P<0.05);serum ornithine,arginine,hippuric acid and proline were significantly upregulated after three months of treatment(P<0.05);serum palmitoleic acid,oleic acid,linoleic acid,palmitoyl carnitine,11Z-octadecenyl carnitine,Cr and uric acid were significantly downregulated after six months of treatment(P<0.05),serum LPC 17:0,LPC 22:4,LPC O-16:0,LPC O-16:1,LPC O-18:0,LPC O-18:1,LPE 18:lsn-1,ornithine,arginine,hippuric acid and phenylalanineproline were significantly upregulated after six months of treatment(P<0.05).②MetPA signal pathways:The signal pathways related to the mechanism of Qilingwenshenxiaonang decoction include linoleic acid metabolism and arginine and proline metabolism.Conclusion:1.The metabolic changes of PCOS patients with phlegm-dampness syndrome were mainly accumulation of FFA and fatty acid amides,reduction of fatty acid β oxidation,phospholipid metabolism and amino acid metabolism disorder,increased androgen synthesis,intestinal flora disorder,and reduced synthesis of bile acids,enhanced glycolysis,inhibited glycogen,abnormal purine metabolism.The abnormalities of alanine,aspartate and glutamate metabolism,arachidonic acid metabolism,arginine and proline metabolism,D-glutamine and D-glutamic acid metabolism,pyruvate metabolism,glycerophospholipid metabolism and aminoacyl-tRNA biosynthesis might involve in PCOS of phlegm-dampness syndrome.Serum uric acid,palmitoyl carnitine,nervonic acid,octadecenoyl carnitine,palmitic acid,glutamic acid,PC 32:1,eicosadienoic acid,eicosatrienoic acid and phenylalanineproline were defined as biomarkers for distinhuishing PCOS patients with phlegm-dampness syndrome from healthy controls.2.Qilingwenshenxiaonang decoction can significantly improve the patient’s clinical symptoms and signs,regulate the patient’s sex hormone disorders and glucose and lipid metabolism disorders.The efficacy of Qilingwenshenxiaonang decoction on treating PCOS patients with phlegm-dampness syndrome after six months is better than three months,and the metabolites changed more significantly.This decoction may promote fatty acid β oxidation,regulate FFA metabolism,phospholipid metabolism and amino acid metabolism,regulate intestinal ecosystems,and promote Cr and uric acid excretion through interventing the signal pathway of linoleic acid metabolism and arginine and proline metabolism.