| Objective To investigate the association between single nucleotide polymorphism(SNP)of transcription factor forkhead box protein 3(FOXP3)gene and the susceptibility to type 2 diabetes mellitus(T2DM)and type 2 diabetic nephropathy(T2DN)in Gansu Han population.Methods Total of 200 patients with type 2 diabetes mellitus(T2DM group)and 200patients with type 2 diabetic nephropathy(T2DN group)were selected as the patient group(Patient group),and 200 healthy subjects as the control group(HC group)from Lanzhou University Second Hospital from March 2021 to April 2022.PCR-HRM and Sanger sequencing were performed to identify the genotypes of FOXP3polymorphisms(rs3761548 C>A,rs2294021 C>T,rs2232365 G>A,and rs3761547T>C).Statistical analysis was calculated by IBM SPSS statistics,chi-square test was used to analyze the Hardy-Weinberg Equilibrium,the distribution differences of alleles and allele frequencies between the case group and the healthy control group,as well as between the T2DM and T2DN groups;using unconditional logistic regression to analyze the association between FOXP3 gene polymorphism and susceptibility to T2DM and T2DN.A gender-stratified evaluation was also conducted.SHEsis online software was used to analyze linkage disequilibrium and construct haplotypes of the four SNPs of the FOXP3 gene.Results(1)The genotype frequencies of the four SNPs conformed to Hardy-Weinberg Equilibrium in the healthy control group.(2)The association analysis of the genotype frequency,and allele frequency distribution,as well as the susceptibility to T2DM and T2DN was as follows:(1)rs3761548 G>A:The genotype AA and allele C,A have significant statistical differences between the patient group and the control group,the T2DM group and the T2DN group(P=0.005,P<0.001;P=0.004,P<0.001).There were statistically significant differences in the distribution of dominant mode[CC vs(CA+AA)],recessive mode[AA vs(CC+CA)],and cumulative mode[CC vs CA vs AA]between the healthy control group and the case group(OR:0.530,95%CI:0.354-0.791,P=0.002;OR:4.897,95%CI:2.398-10.003,P<0.001;OR:4.897,95%CI:2.398-10.003,P<0.001;P=0.005).While between the T2DM group and the T2DN group,the distribution of dominant mode[CC vs(CA+AA)],recessive mode[AA vs(CC+CA)],overdominant mode[CA vs(CC+AA)]and cumulative mode[CC vs CA vs AA]were statistically significant(OR:0.648,95%CI:0.425-0.987,P=0.043;OR:4.030,95%CI:2.272-7.151,P<0.001;OR:0.422,95%CI:0.232-0.768,P=0.005;P<0.001).(2)rs2294021 C>T:There was a statistical difference in the distribution of genotype TT between the control group and the patient group,while there was no significant difference in the distribution of allele C and T(P=0.005,P=0.121).The distribution differences of genotype TT and alleles C and T were both statistically significant between the groups of T2DM and T2DN(P=0.002,P=0.002).The distribution of recessive mode[TT vs(CC+CT)],overdominant mode[CT vs(CC+TT)]and cumulative mode[CC vs CT vs TT]were statistically different between the control group and the patient group(OR:1.543,95%CI:1.078～2.208,P=0.018;OR:0.540,95%CI:0.360～0.810,P=0.003;P=0.005).The distribution of recessive mode[TT vs(CC+CT)],overdominant mode[CT vs(CC+TT)]and cumulative mode[CC vs CT vs TT]were statistically different between the groups of T2DM and T2DN(OR:1.946,95%CI:1.299-2.914,P=0.001;OR:0.483,95%CI:0.281-0.831,P=0.009;P=0.002).(3)rs2232365 G>A:There were statistically significant differences in genotype AA and alleles G and A between the patient group and the control group(P<0.001;P<0.001).The distribution of genotype AA showed statistical differences between the T2DM and T2DN groups,while there was no significant difference in alleles G and A(P=0.005,P=0.706).A statistical difference was shown in the distribution of recessive mode[AA vs.(GG+GA)]and cumulative mode[GG vs.GA vs.AA]between the groups of control and the patient(OR:3.244,95%CI:1.847-5.695,P<0.001;P<0.001).The distribution of overdominant mode[GA vs(GG+AA)]and cumulative mode[GG vs GA vs AA]was statistically different between the groups of T2DM and T2DN(OR:0.420,95%CI:0.246～0.717,P=0.001;P=0.005).(4)A significant statistical difference was shown in the distribution of genotype CC between the T2DM group and the T2DN group(P=0.015).There was a significant difference in the distribution of recessive mode[CC vs(TT+TC)]and cumulative mode[CC vs TC vs CC]between the groups of T2DM and T2DN(OR:3.162,95%CI:1.127-8.874,P=0.029;P=0.015).(3)Further stratified analyze based on gender differences showed that in female subjects,the risk of rs3761548 increased around 3-fold and 4.5-fold with T2DM and T2DN,respectively,while there was no significant correlation with rs2294021.In males,the rs3761548 polymorphism increased the susceptibility to T2DM and T2DN around 5.4-fold and 3.4-fold,respectively,while the rs2294021 polymorphism increased the susceptibility to T2DN nearly 2-fold.The AA genotype at the rs2232365 polymorphism increased the risk of developing T2DM and T2DN in women by approximately 6-fold and 2-fold,respectively,while men carrying the AA genotype have a 2-fold increased risk of T2DM.Male T2DM patients carrying the C allele at the rs3761547 locus may be more inclined towards the development of kidney disease.(4)Linkage disequilibrium analysis:In the patient group,there was a linkage disequilibrium between rs3761548and rs2232365(D’=0.227,r~2=0.007),rs3761548 and rs3761547(D’=0.358,r~2=0.005),rs2294021 and rs3761547(D’=0.286,r~2=0.009).While there is no linkage disequilibrium between rs3761548 and rs2294021(D’=0.136,r~2=0.007),rs2294021and rs2232365(D’=0.132,r~2=0.007),rs2232365 and rs3761547(D’=0.058,r~2=0.001).In the healthy control group,there was a linkage disequilibrium between rs3761548and rs2294021(D’=0.320,r~2=0.012),rs3761548 and rs2232365(D’=0.277,r~2=0.043),rs2294021 and rs2232365(D’=0.572,r~2=0.102),rs2294021 and rs3761547(D’=0.403,r~2=0.015),rs2232365 and rs3761547(D’=0.249,r~2=0.027),while there was no linkage disequilibrium between rs3761548 and rs3761547(D’=0.000,r~2=0.000).Between the case group and the healthy control group,there was a linkage disequilibrium between rs3761548 and rs3761547(D’=0.301,r~2=0.003),rs2294021and rs2232365(D’=0.219,r~2=0.017),rs2294021 and rs3761547(D’=0.307,r~2=0.010),while between rs3761548 and rs2294021(D’=0.141,r~2=0.005),rs3761548 and rs2232365(D’=0.024,r~2=0.000),rs2232365 and rs3761547(D’=0.110,r~2=0.004)had no linkage disequilibrium.(5)Haplotype analysis:There were significant differences in the distribution of ACGT,ATAT,ATGT,CCGC,CCGT,CTAC and CTAT haplotype frequencies in the control group and patient group(P<0.05).It is suggested that haplotype ACGT(OR:3.650;95%CI:2.097～6.351),ATGT(OR:9.579;95%CI:3.102～29.575),CCGC(OR:2.980;95%CI:1.291～6.877),CTAT(OR:1.878;95%CI:1.187～2.972)may be the risk factors of T2DM,while haplotype ATAT(OR:0.577;95%CI:0.330～1.007),CCGT(OR:0.332;95%CI:0.255～0.433)and CTAC(OR:0.295;95%CI:0.127～0.689)may be the protective factors of T2DM in Gansu Han population.Conclusion Polymorphisms of the FOXP3 gene(rs3761548 C>A,rs2294021 C>T,rs2232365 G>A,and rs3761547 T>C)may be associated with susceptibility to T2DM and T2DN in Gansu Han population. |
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