Metabolomic Study On Biomarker Discovery And Pathogenesis Mechanism Of Pelvic Organ Prolapse

Objectives:Previous studies suggest the existence of many differentially expressed genes and proteins involved in POP Metabolites are the end products of cellular regulatory processes,and their levels can be regarded as the ultimate response of biological systems to genetic or proteinic changes.So it will be informative to investigate the potential machanisms and biomarkers of POP by metabolomics.Methods:We applied a non-targeted metabolomics approach using ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UHPLC-Q-TOF-MS),on and above the POP group II degree as well as the control group,serum and urine samples were detected for endogenous small molecule metabolites,combined with chemometrics and multiple statistical analysis to find differences in the metabolic production of POP development related,finally the existence of significant differences metabolites,ROC curve analysis and primary cultured vaginal wall fibroblasts as experimental model in vitro,the original generation of vaginal wall fibroblast cultivation,SABC method was used to identify vimentin,keratin and smooth muscle actin by immunocytochemical staining.In addition,these functions will be examined afterover express or silence related metabolic genes for further validate the results of metabolomics.Results:1.Metabolomics study of serum samples from patients with POP(n = 24)and controls(n = 22)revealed a total of 59 metabolites that are significantly different between the two groups.2.Between urine samples from POP patients(n = 45)and controls(n= 59),33 metabolites differed significantly.3.Metabolic pathways affected by these differentially expressed metabolites were analyzed.In both serum and urine samples,three pathways including arginine biosynthesis and purine metabolism were found to be significantly related to POP.4.Six metabolites including GPC,1-methyladenosine,maleic acid,Lpyroglutamic acid,inosine,and citrate are significantly changed in both serum and urine samples from patients with POP.Receiver operating characteristics(ROC)curve analysis showed that using these six metabolites as a biomarker could distinguish patients with POP from controls with good accuracy in both serum(AUC=1)and urine samples(AUC=0.854).5.Cultivation from POP vaginal wall tissue of patients with success in the generation of vaginal wall fibroblasts,and USES the SABC method to identify the immunocytochemistry staining,further study found that patients with POP raised differences in serum metabolites phosphatidyl inositol vaginal wall fibroblasts induced matrix metalloproteinase-1(MMP-1)mRNA expression levels of tissue matrix metalloproteinases inhibitor-1(TIMP-1)mRNA expression levels.Conclusions:we were able to develop a six-metabolite panel consistingof GPC,1-methyladenosine,maleic acid,L-pyroglutamic acid,inosine,and citrate for diagnosis of POP.these results further extended our understanding of key regulatory metabolic pathways involved in the pathophysiology of POP,as well as provided some promisingbiomarkers for effective POP diagnosis.