Study On The Correlation Between Methylation Of Carbohydrate Response Element Binding Protein(ChREBP) Gene Promoter And Type 2 Diabetes Mellitus In Peripheral Blood Mononuclear Cells

Objective This research aims at investigating the methylation level of promoter region of ChREBP gene in patients with type 2 diabetes mellitus(T2DM)and peripheral blood mononuclear cells.The relationships between methylation of ChREBP promoter and T2DM will be studied.Eventually,the effect of cisplatin on the expression of ChREBP could provide new evidence for the molecular mechanism which regulates the development and progression of early diabetes mellitus.Method According to the 1999 WHO diagnostic criteria for diabetes mellitus,we selected the Department of Wuhan university zhangnan hospital Endocrinology was diagnosed as 2 Type of diabetes in 65 cases and the same period the hospital health examination in 25 cases.The general clinical baseline data,including sex,age,height,weight,waist circumference,hip circumference,blood pressure,duration of diabetes mellitus,were calculated according to the formula,group A(control group)and group B(diabetes mellitus)Waist to hip ratio(WHR),body mass index(BMI).(FPG),serum cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),high cholesterol(LDL-C),high cholesterol(LDL)and low density lipoprotein cholesterol(LDL-C)were measured by automatic biochemical analyzer.(HDL),homocysteine(HCY),glycosylated hemoglobin(HbAlc)and other biochemical markers;extraction of peripheral blood leukocyte DNA,the application of methylation-specific PCR bisulfite restriction endonuclease method(BSP)to detect the methylation level of ChR.EBP gene promoter.The mRNA expression of ChREBP gene was detected by RT-PCR.SPSS17.0 statistical software for data processing.Measurement data with mean ± standard deviation,said the two groups were compared between the t test.Methylation rate between the two groups using the chi-square test to P<0.05 for the difference was statistically significant.Results Compared with the normal control group,there was no significant difference in age,gender ratio,SBP,DBP and WHR in T2DM group(P>0.05),which indicates that the baselines were at the same level and the data were comparable(P<0.05).The mean methylation rate of T2DM group was(42.48%),while the average methylation rate in the normal group was(5%),and the mean methylation rate in the T2DM group was higher than that in the normal control group.The rates of methylation of 8 CpG sites in the ChREBP gene were higher than that in the normal control group:CPG sites 1,CPG sites2,CPG sites3,CPG sites4,CPG,CPGsites6,CPGsites7and CPGsites8.There were significant differences between the two groups(P<0.01).MRNA expression of ChREBP gene was detected by RT-PCR method.The results showed that the expression of ChREBP gene in type 2 diabetes group was significantly higher than that in control group(mRNA).Conclusion The ChREBP gene promoter methylation level in patients with Type 2 diabetes mellitus was significantly increased,affecting the expression of ChREBP gene.In this study,I investigated the methylation status of CpG site of ChREBP gene promoter,suggesting that the abnormal methylation level of ChREBP gene in peripheral blood leukocytes of type 2 diabetic patients may be related to the occurrence of diabetes mellitus.