The Association Of Adiponectin Gene SNP+45, SNP+276 With Metabolic Syndrome And Plasma Adiponectin Concentration

ObjectivesThe purpose of this study was to investigate the genotype distribution of adiponectin gene SNP+45 and SNP+276 T/G polymorphisms,analyse the relationship between adiponectin gene SNP+45,SNP+276 and MS or plasma adiponectin concentration and provide a theoretical basis,valuable genetic markers for MS.Subjects and Methods1.SubjectsAll subjects are Han peoples of Shandong.The metabolic syndrome(MS) patients were recruited from the old people.200 control subjects were selected from healthy old people matching with sex and age.2.Extraction of DNAThe DNA was extracted with the TIANamp Genomic DNA Kit.3.Analysis of adiponectin gene SNP+45 and SNP+276 polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)Gene-specific primers of adiponectin gene exon 2 and intron 2 for polymerase chain reaction was designed using Oligo 6.0 software and adiponectin DNA sequence recorded by NCBI online.Restriction endonuclease Sma I and EcoR I was used to determine the genotype of SNP+45 and SNP+276.4.Assay of plasma adiponeetin concentrationPlasma adiponectin levels were measured with ELISA method.Results 1.Clinical characteristics of the MS group and control groupThere were no significant differences in age and gender between MS group and control group(P＞0.05)).Compared with controls,body mass index(BMI),systolic blood pressure(SBP),diastolic blood pressure(DBP),wist circumference(WC),hip circumference(HC),wist-to-hip ratio(WHR),uric acid(UA),triglycerides(TG),fasting blood glucose(FBG) were significantly higher in the patients with MS(all P＜0.05),whereas high-density lipoprotein cholesterol(HDL-c) was significantly lower in MS group(P＜0.05)).2.Genotype distribution and allele frequencies of MS and control groups2.1 SNP+276 siteThe TT,TG and GG genotype frequencies of MS group were 4.46%,37.05% and 58.48%respectively;The frequencies in control group were TT 11%,TG 37.5%,GG 51.5%respectively.The frequency of GG genotype in MS group was significantly higher,but TT genotype frequency was significantly lower than that of control group.Statistical difference was found in genotypic frequency between the two groups(P=0.031).T and G allele frequencies were 22.99%,77.01%in MS group and 29.75%,70.25%in control group respectively,there were significant difference between the two groups(P=0.025).2.2 SNP+45 siteThe TT,TG and GG genotype frequencies were52.68%,41.07%,6.25%in MS group,and TT54%,TG40%,GG6%in control group respectively.There was no statistical difference in genotypic frequency between these two groups(P＞0.05).T and G allele frequencies were73.21%,26.79%in MS group and 74%,26%in control group respectively,there was no significant difference between the two groups(P＞0.05).3.Comparison of clinical and laboratory characteristics between MS and control group3.1 SNP+276 The levels of WC,HC in MS group GG genotype were significantly higher than those in non-GG genotype,but the levels of UA in GG genotype was significantly lower than that in non-GG genotype,there were significant statistical significance(P＜0.05).In control group,the levels of HC, cholesterol(CH),γ-GT,PLT in GG genotype were significantly higher than those in non-GG genotype,but the levels of WHR,UA were significantly lower than those in non-GG genotype(P＜0.05).3.2 SNP+45 The level of PLT in MS group TT genotype was significantly lower than that in non-TT genotype.In control group,the levels of HC,ALT in TT genotype were significantly higher than those in non-TT genotype,but the levels of WBC,PLT were significantly lower than those in non-TT genotype(P＜0.05).4.The relationship between SNPs and MS4.1 Logistic Regression Model of MS risk factorsStepwise multivariate logistic regression analysis was performed to determine the predictive variables of MS among+45 T/G and+276 G/T polymorphism,age, sex,blood pressure,fasting blood glucose,BMI,which showed that BMI,DBP,FPG, TG,+276G/T polymorphism were the independent predictors.Carriers with GG genotype had higher risk of MS than the carriers with TT genotype.4.2 The affect of SNP+45 T/G and SNP+276 G/T polymorphisms on the components of MSStepwise multivariate logistic regression analysis was used to determine the predictive variables of MS among+45 T/G and+276 G/T polymorphisms,blood pressure,sugar,BMI,CH,TG,HDL-c,LDL-c for the components of MS,which showed that SNP+276 site G allele increased the risk of high TG and FPG.But+45 T/G polymorphisms were not the significant predictors of the components of MS.5.Plasma adiponectin levels5.1 The plasma level of APN in MS group was significantly lower than that in control group.5.2 The plasma level of APN in women was significantly higher than that in men.5.3 According to the cutoff point of components in MS,the group was divided into two subgroups,the plasma levels of APN showed no significant difference between the normal TG group and the high TG group,normal blood pressure group and high blood pressure group.But the plasma levels of APN in normal HDL-c group,normal BMI and normal FBG were significantly higher than those in low HDL-c group,high BMI group and high FBG group respectively.5.4 In the SNP+45 site,the plasma levels of APN in TT genotype and non-TT genotype were 12.87±2.42ug/ml,13.78±2.07ug/ml respectively.The level of APN in TT genotype was significantly lower than that in non-TT genotype.In the SNP+276 site,the plasma levels of APN in GG genotype and non-GG genotype were 13.24±2.30ug/ml,13.38±2.32ug/ml respectively.There was no significant difference between the two genotypes.5.5 Plasma adiponectin concentration was negatively correlated with waist circumference,waist-hip ratio and positively correlated with HDL-c. Conclusions1.There were T/G polymorphisms of apM1 gene +45 site and +276 site in Shandong population.2.The G/T polymorphism of +276 site was associated with the metabolic syndrome.Subjects with the GG genotype at +276 site had a significantly increased risk of the metabolic syndrome,G allele increased the risk of high TG and FPG.3.The adiponectin concentration in MS group was markedly lower than that in control group.The adiponectin concentration in women was markedly higher than that in men.HDL-c,WC,WHR were independent factors of adiponectin concentration.4.SNP+45 T/G polymorphism associated with the level of APN.The APN level of TT genotype was significantly lower than that in non-TT genotype(P＜0.05).5.We didn't find the relationship between SNP +45 site genotype and the incidence of MS,and the association of SNP +276 G/T polymorphism and the level of APN didn't be found.