Study On Level Of Bone Morphogenetic Protein-9 In Polycystic Ovarian Syndrome And The Relationship With Insulin Resistance

Objective To investigate the changes of BMP-9 in Polycystic Ovary Syndrom(PCOS)and its relationship with insulin resistance(IR)and glycolipid metabolism.Methods(1)High-fat feed and Dehydroepiandrosterone(DHEA)were subcutaneously injected into PCOS model rats,and then fed with ordinary feed,treated with gastric perfusion of metformin(MF),subcutaneous injection of Exenatide(EX).The levels of insulin resistance and liver glucose metabolism were evaluated by Euglycemic-Hyperinsulin Clamp(EHC)and 3-H tracer technique.Levels of sex hormones,serum glucose and lipid were measured.(2)The rat serum,ovaries and liver were retained,the serum BMP-9 levels measured with ELISA,BMP-9 changes in rat ovaries were observed by immunohistochemistry(IHC).The protein expressions of BMP-9,AKT,insulin receptor(IR)and Androgen Racceptor(AR)in the ovaries and the livers were determined by Western Blotting(WB).The gene expression of insulin like growth factor-1(IGF-1),PEPCK and G-6-P in the livers were determined by RT-PCR.(3)PCOS patients were selected,and normal controls and insulin resistance patients were involved in.PCOS patients were treated with Exenatide for 3 months.All subjects were given EHC to evaluate the degree of IR,the levels of BMP-9 in each subjects were determined.The correlation between BMP-9 and clinical indicators was studied.Results(1)The weights,blood pressures,levels of serum glucose and lipid of the rats with high fat diet and PCOS model increased significantly(P<0.01).The levels of MF and EX group rats were significantly decreased(P<0.01).During the clamp,basal serum glucose and FFA in HFD and PCOS rats increased significantly(P<0.01),while those in ND,MF and EX groups were decreased significantly(P<0.01).Concentrations of serum glucose and lipids were significantly lower than those in the basal state(P<0.01).The concentrations of insulin at steady-state were significantly increased in HFD and PCOS groups than at basal state(P<0.01),and in group ND,MF and EX significantly decreased(P<0.01).GIR and GRd in group HFD and PCOS groupdecreased significantly(P<0.01,P<0.05)while increased in MF and EX.HGP significantly increased(P<0.05),while decreased in MF and EX group(P<0.05).SUPP HGP significantly decreased(P<0.01,P<0.05)while increased in group MF and EX.(2)Serum BMP-9 levels decreased in HFD and PCOS group rats,increased in MF and EX group rats(P<0.05).The numbers of granulosa cells identified by anti BMP-9 antibody in the ovarian tissues of group HFD and PCOS rats decreased,and increased in group ND,MF and EX groups.The Western blotting results of rat ovarian tissue showed that the expression of BMP-9 and p-AKT proteins in the ovaries of HFD and PCOS model rats decreased significantly(P<0.05),group ND,MF group and EX group increased significantly((P<0.05)),and the expressions of AR and p-IR proteins in group HFD and PCOS model rats increased,the expressions of ND group,MF group and EX group significant decreased(P<0.05).The results of Western blotting in rat liver tissues were similar to those in the ovaries.The expressions of IGF-1 m RNA expression in PCOS group increased(P<0.05),ND,MF and EX group significantly reduced(P<0.05).Compared with group NC,PCOS group PEPCK m RNA increased.G-6-P m RNA in group PCOS were increased(P<0.05),and in group ND,MF and EX significantly decreased(P<0.05).(3)the body weight of PCOS patients decreased significantly(P<0.01),M-value,levels of BMP-9 significantly increased(P<0.01).Levels of BMP-9of IR and central obesity patients were significantly lower than non IR and non central obesity patients(P<0.01).Correlation analysis showed that BMP-9 was significantly negatively correlated with weight,BMI,waist circumference,hip circumference,waist hip ratio,OGTT 0 'blood glucose,OGTT 60' blood glucose,OGTT 2H blood glucose,OGTT 0 'insulin,OGTT 2H insulin,triglyceride,total cholesterol,low density lipoprotein cholesterol,and a significant positive correlation with M value and progesterone.With BMP-9 as the dependent variable and its related other indexes as independent variables,multiple linear stepwise regression analysis is carried out.Only the M value is included in the regression equation,and Y(BMP-9)=19.667+2.206X(M).Conclusion(1)Exenatide can reduce body weight and insulin resistance in PCOS rats,and improve liver glucose metabolism and glucose and lipid metabolism disorders.(2)Serum BMP-9 levels and the protein expressions decreased in livers and ovaries of PCOS rats.It may be involved in glucose and lipid metabolism and PI3K/AKT insulin signaling pathway in PCOS rats.(3)The serum BMP-9 levels of PCOS patients with insulin resistance were decreased,and that of BMP-9 increased after treatment.BMP-9 was an independent influence factor of insulin resistance in PCOS patients with insulin resistance.