| [Objective]To investigate the genotype and allele frequency distribution characteristics of klotho gene G395A,F352V,and C370S single nucleotide polymorphisms(SNPs)among type 2 diabetes patients and a healthy population of the Yi population in Chuxiong area,to investigate the level of klotho protein and the correlation between gene polymorphisms and type 2 diabetes(T2DM).[Methods]?This study selected yi nationality residents with age over 20 in Chuxiong,Yunnan Province,China as research.The people without cross racial marriage within three generation were recruited as the study participants.They were divided into healthy control group(Group A)and T2DM group(Group B).The average age of Group A with 120 paticipants(82 males and 38 females)was 52.22 ± 5.63 years;there were 128 patients(73 males and 55 females)with an average age of 55.48 ±7.18 years in T2DM group.?The SNPs of three loci of kotho gene were detected by real-time fluorescence quantitative PCR(FQ-PCR),The frequencies of genotype and allele distribution were compared between the two groups.? Olympas AU5400 automatic biochemical analyzer measures serum creatinine(Scr),urea nitrogen(BUN),uric acid(UA),total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),High density lipoprotein cholesterol(HDL-C)detection;glucose oxidase method to measure fasting plasma glucose(FBG)and postprandial two hours blood glucose(2hPBG);latex agglutination reaction method for the determination of glycated hemoglobin(HbA1C);The fasting insulin level(FINS)was measured by radioimmunoassay and the plasma klotho level was measured by ELISA.Compare differences between different genotypes and diabetic and non-diabetic groups;?The ?2test was used to analyze the difference of genotype and allele frequency of klotho gene G395A between two groups;Independent sample t-test was used to analyze the relationship between plasma klotho levels and type 2 diabetes mellitus;Logistic analysis was used to analyze the influencing factors of T2DM.[Results]?The Klotho gene G-395A distribution frequency,GG genotype frequency was 68.5%;GA+AA genotype frequency was 31.5%;allele G and A frequency were 83.8%and 16.2%,respectively.?Klotho gene F352V,C370S polymorphism can not be verified,and all samples were wild type(FF,CC type);?The G/G gene frequency and G allele frequency of Klotho gene G395A were higher in group B than the frequencey of group A(?2?5.107,P = 0.023;?2 4.102,P = 0.043).?Plasma klotho levels(ng/ml)were lower in the type 2 diabetes group(8.66 ? 2.04)than in the healthy control group(11.80 ± 1.69),P<0.01.? Plasma Klotho levels were negatively correlated with age,SBP;DBP,FBG;TC,TG,LDL-C,BUN,Scr,UA(r =-0.164,-0.297,-0.191,-0.560,-0.396,-0.409,respectively).-0.416,-0.269,-0.325,-0.271,P all<0.01),? Logistic regression showed that the occurrence of type 2 diabetes mellitus was positively correlated with FBG,TG,and LDL-C,and negatively correlated with klotho protein level(OR values were 37.013,6.852,41.261,0.259,P<0.05),and G/A with G395A.G genotype has nothing to do(P>0.05).[Conclusion]?The Klotho gene G395A polymorphism existed in the Yi population in Chuxiong area.G/G gene frequency and G allele frequency of Klotho gene G395A were higher in T2DM group than in the healthy control group;? There was no polymorphism in the F352V and C370S loci of Klotho gene.?Plasma klotho levels in patients with T2DM were lower than those in healthy controls.Logistic analysis indicated that susceptibility of T2DM was positively correlated with FBG,TG,and LDL-C,and negatively correlated with the level of klotho protein?and Klotho may be a protective factor for T2DM.[Objective]To study the genotype and allele frequency distribution of Klotho gene G395A single nucleotide polymorphism among healthy population,T2DM without lower extremity atherosclerosis(LEA),T2DM with LEA in Chuxiong area;To investigate the association between klotho protein level and gene polymorphisms in T2DM with LEA of Yi population in Chuxiong area of Yunnan Province.[Methods]? A case-control study was performed about the arteries of both lower extremities using the Aloka Alpha Doppler Color Doppler Ultrasonography to the participants.?The subjects were divided into healthy control group(Group A)of 120 patients(82 males and 38 females)with an average age of 52.22 ±5.63 years,61 patients(40 males and 21 females)with T2DM(Group B1)of mean age 53.79 ± 6.59 years,and T2DM combined with LEA group(Group B2)of 67 cases(33 males and 34 females),with an average age of 57.01 ± 7.39 years.Plasma klotho protein levels and klotho gene G395A genotype,allele distribution frequency were compared among the three groups ? The ?2 test was used to analyze the genotype and allele frequency distributions of the Klotho gene G395A among the three groups.The one-way analysis of variance(ANOVA)was used to correlate plasma Klotho level with frequency of T2DM with LEA;Logistic analysis showed that influencing factors of T2DM combined with LEA rigidification.[Results]? The genotype and allele frequency distribution of Klotho gene G395A was not significantly different among the three groups(?2=5.552,P>0.05;?2=4.289,P>0.05);? Plasma Klotho concentrations in T2DM combined with LEA group and T2DM group were lower than those in normal control group,and plasma Klotho levels in T2DM combined with LEA group were lower than those in type 2 diabetes mellitus group(7.61 ± 1.84 vs 9.82 ± 1.58)(vs 11.80 ± 1.69).By covariance analysis,the difference in klotho concentration remained remarkedly among the three groups(p<0.01);? Linear correlation analysis Plasma klotho protein levels in the Yi population were negatively correlated with age,SBP,DBP,FBG,TC,TG,LDL-C,BUN,Scr,UA,2hPBG,HBAlC,and sclerosis score(r =-0.164,-0.297,respectively).-0.191,-0.560,-0.396,-0.409,-0.416,-0.269,-0.325,-0.271,-0.253,-0.198,-0.530,P all<0.05);? Multivariate stepwise regression analysis showed that age and arteriosclerosis scores are independently related factors affecting plasma Klotho levels;?Logistic regression showed that susceptibility of diabetic LEA was associated with FBG,TG,LDL-C,Scr,and klotho proteins(OR values 1.372,1.827,4.206,1.054,0.401,P<0.05).[Conclusion]? The concentration of plasma klotho in T2DM combined with LEA group was less than that of T2DM group and normal control group gradually,which was negatively correlated with the lo?wer extremity arteriosclerosis score?Klotho protein may have a protective effect on atherosclerosis;? Klotho G395A polymorphism is not associated with the onset of atherosclerosis in type 2 diabetes with lower extremity. |
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