Association Of The Rs2268388 Polymorphism Of ACACB Gene With Diabetic Mellitus And Diabetic Kidney Disease

Part 1:The Experimental Conditions for ACACB gene rs2268388 Polymorphism DetectionObjective:To establish a rapid and specific quantitative PCR assay for detecting of ACACB gene rs2268388 polymorphism.Methods:The primers and Taqman probe targeted at signature sequence of ACACB gene rs2268388 polymorphism were designed and applied to real time PCR assays. The different concentrations were set up, and the sensitivity and specificity were evaluated.Result:Initial denaturation time of PCR was 10min; annealing and extending time were lmin; probe concentration of Taqman PCR was 0.5ul (0.2uM), TaqMan Genotyping Master Mix was 5ul, dH2O was 2.5ul,total volume was 10ul. and the result of ACACB gene rs2268388 polymorphism showed the high specificity of the reaction.Conclusions:the assay was sensitive and specific for rapid identification and quantitative detection of ACACB gene rs2268388 polymorphism. It will be an effective method for the surveillance and diagnosis of ACACB gene rs2268388 polymorphism.Part 2:The Polymorphism of ACACB gene Related to Type 2 diabetes mellitusObjective:To investigate the relationship of rs2268388,rs2268393 polymorphism of ACACB gene with Type 2 diabetes mellitus (T2DM) in Kunming Han Chinese.Methods:rs2268388 and rs2268393 polymorphism of ACACB gene were detected respectively by Taqman PCR(Three-star) and PCR-RFLP in 184 untreated Kunming han patienes with type 2 diabetes mellitus (T2DM) and 70 normal controls (NC). The genotype, allele frequency, risk factors and related clinical and biochemical data were compared among groups.Results:(1)There were no significant differences in rs2268393 genotype and allele frequency between T2DM group and NC group (χ2=4.810,p=0.09, χ2=1.29,p=0.24); C/C genetype frequency of rs2268388 in T2DM group was significantly higher than those in NC group(χ2= 10.469, p=0.005); C allele frequency of rs2268388 in T2DM group was significantly higher than those in NC group (χ2=4.71, p=0.007). (2) Logistic regression analysis showed that in patients with T2DM, rs2268388 C/C genotype and higher hepatic triglyceride (TG) were risk factors for occurrence of T2DM.Conclusions:rs2268388 C/C genotype of ACACB gene may be associated with T2DM in Kunming Han Chinese.Part 3:Association of The rs2268388 Polymorphism of ACACB gene With Diabetic kidney diseaseObjective:To explore the association of the rs2268388 polymorphism of ACACB gene with Diabetic kidney disease (DKD) in Kunming Han Chinese.Methods:The genotype was detected by Taqman PCR(Three-star) in 248 Kunming han patients with type 2 diabetes mellitus (T2DM),including 85 normal albuminuria (DKDo group),99 microalbuminuria (DKD1 group),64 macroalbuminuria (DKD2 group), and 70 normal controls (NC). The genotype and allele frequencies and relative clinic data were compared among groups.Results:(1) C/T genotype frequencies in DKD0、DKD1 and DKD2 group were significantly lower than those in NC group (Z=-2.391, P=0.017); T/T genetype frequencies in DKD] group and DKD2 group were significantly higher than those in DKDo group(Z=-2.522, P=0.012;Z=-4.981, P=0.001) T/T genetype frequencies in DKD2 group were significantly higher than those in DKD1 group and NC group(Z=-4.981, P=0.001), but nor in allele frequencies among the four groups(x2=0.099, P=0.753). (2) Logistic regression analysis showed that in patients with T2DM, diabetic duration, rs2268388 T/T genotype, higher blood triglycerides(TG), higher Glycated hemoglobin Alc (HbAlc%) and higher uric acid (HUA) hematic disease were risk factors for the occurrence of DKD.Conclusions:rs2268388 T/T genotype of ACACB gene may be related with DKD in Kunming Han Chinese.