| Objective: This study was designed to explore the metabolic pathways of PCOS by detecting blood and urine samples from rats with polycystic ovary syndrome(PCOS)using nuclear magnetic resonance spectroscopy(NMR)detection techniques,and to search for potential biomarkers of PCOS to construct PCOS.Early diagnosis.Methods: Thirty-three SD rats aged 23 days were used to construct PCOS model with dehydroepiandrosterone(DHEA).Rats were randomly divided into 2 groups,group A PCOS model group and group B oil control group.Group A rats were given DHEA(6mg/100g)daily for 20 days,group B rats were given 0.2ml of oil,and the last injection was fasted.The urine was collected in a sodium azide solution collection bottle and centrifuged at low temperature.The supernatant was collected in a cryotube,stored at-80 ° C,and the rats were fixed under isoflurane anesthesia.Blood was collected from the tail vein blood.The blood was collected in a centrifuge tube,centrifuged,and the supernatant was collected.Store at-80 ° C.HE staining was performed on one side of the ovary,and the other side was placed in a cryotube and stored at-80 °C.The serum was taken for sex hormone and insulin detection,and the urine and the urine were separately analyzed by NMR.Result: 1.Results of PCOS model: The results of HE staining of ovaries in the two groups were observed.It was found that the follicles of ovarian cystic dilatation increased in the PCOS group,the number of follicular granulosa cells decreased,and no dominant follicles were observed,while the control group observed different developmental stages.In the follicles,multiple layers of granule cells are visible in the follicle,and dominant follicles are visible.The ovaries,uterus and body weight of the two groups were compared.It was found that the weight of the PCOS group was heavier than that of the control group(P<0.05),but there was no significant difference between the two groups(P>0.05).Compared with the control group,the levels of androgen(T)and luteinizing hormone(LH)were higher than those in the control group,but there was no significant difference in LH,and the level of estrogen(E2)in the PCOS group was increased(P<0.05)..Compared with fasting blood glucose(FPG)and fasting insulin(INS),the levels of INS in PCOS group were decreased(P<0.05),FPG and HOMA-IR levels were increased,but FPG and HOMA-IR were not statistically significant.difference.2.Serum NMR analysis results: 70 metabolites were identified in the two groups.Compared with the control group,the serum metabolites in the PCOS model group were found to have differences in 31 metabolites(P<0.05).Significantly different metabolites were creatine,proline,and isoleucine(P < 0.01).The serum levels of glucose and pyruvic acid in the PCOS group were significantly higher than those in the control group,and the intermediate metabolites(citrate)in the tricarboxylic acid cycle were significantly different(P<0.05).From the perspective of lipid metabolism,the levels of cholesterol,lipid,glycerol,acetoacetic acid and β-hydroxybutyrate in the PCOS group were significantly higher than those in the control group(P<0.05).It can also find the levels of raw sugar amino acids(alanine,glutamic acid,glutamine,proline,aspartic acid,asparagine,threonine)and essential amino acids(isoleucine,leucine,The levels of proline,lysine and methionine were higher than those of the control group,indicating that amino acid catabolism and anabolism were abnormal,while arginine,phenylalanine and tyrosine levels were decreased,and the difference was statistically significant(P< 0.05).In addition,the levels of dimethylamine,trimethylamine,creatine,and acetyl-glycoprotein were found to be higher than those in the control group,and the difference was statistically significant(P<0.05).3.Urine NMR results: 32 metabolites can be found to be different,among which there are significant differences in metabolites such as choline,glucose,dimethylamine,aspartic acid,inositol,isoleucine,styrene Amino acid,valine,acetoacetic acid,creatinine,trimethylamine,glycine,glycerol + lipid,glycoprotein,creatine(P <0.01).Compared with the control group,the levels of glucose and intermediate metabolites in the urine of the two groups were higher than those in the control group(P<0.05).The levels of choline,glycerol and ketone in the lipids were glycerol + lipid,acetoacetate,choline,The level of 3-hydroxybutyrate was elevated(P<0.05).For amino acid levels,elevated levels of various amino acids(isoleucine,leucine,valine,alanine,glycine,phenylalanine)were found(P < 0.05).In addition,trimethylamine,creatine,creatinine,glycoprotein,and inositol levels were found to be elevated(P < 0.05).Conclusion:1.There is a metabolic difference between PCOS and normal rats,and metabolic disorders occur in carbohydrates,lipids and amino acids.2.There is also a difference in metabolism between different body fluids in blood and urine of PCOS rats. |
