Study On The Changes Of Telomere Length And Telomerase Activity In Peripheral Blood Leukocytes Of Patients With Diabetes Mellitus

Background: Diabetes mellitus is a metabolic disease characterized by high blood glucose,and if the long-term maintenance of hyperglycemia in the body can cause a series of complications,such as kidney,liver,heart,blood vessels,eyes and other damage.So diabetes has a very serious impact on human health.The incidence of diabetes is increasing year by year,and there is no effective medical treatment to cure this disease in the world.There is no characterization of the initial diabetes,but when the patient to the hospital,most of the islet cells have lost their function,and they can only rely on drugs to maintain blood glucose balance.Early detection and early treatment of diabetes has become a hotspot of the research.We detect telomere length and telomerase activity in peripheral blood leukocytes by means of molecular biology,and using this method to study the relationship between the changes of telomere length and telomerase activity in patients with diabetes mellitus.Method:The extraction of 30 cases of type 1 diabetes mellitus(T1DM),60 cases of type 2 diabetes mellitus(T2DM),40 cases of type 2 diabetes with atherosclerosis(DAS)in patients with peripheral white blood cell genomic DNA.Using the real-time fluorescence quantitative PCR method to test the length of telomere.And using the 36B4 gene as the control,detection of different types of diabetes and telomere length changes in diabetic patients and normal human.So we can explore the effect of diabetes on human peripheral blood leukocyte telomere length and using multiple linear regression analysis to explore the impact of specific factors of telomere length.The peripheral blood white blood cells and plasma were extracted from 40 patients with T1 DM and 80 patients with T2 DM,and the presence of telomerase activity in leukocytes and plasma was detected by TRAP-PCR;reuse of fluorescent quantitative PCR method for the detection of telomerase activity;so as to explore the effect of the telomerase activity in peripheral blood leukocytes and plasma of diabetic patients,and then use multiple linear regression analysis to explore the main factors that influence the activity of telomerase.Results:It establishes a reliable method for the detection of telomerase activity and telomere length.The telomere length of peripheral blood leukocytes in patients with T1 DM,T2DM and DAS were all less than that of healthy control(NC)group(P<0.05);in the different types of diabetes,the T1 DM group was shorter than the DAS group and the T2 DM group,the DAS group was shorter than the T2 DM group.By using multivariate linear regression analysis obtained: In the T1 DM group,the age was negatively correlated with the length of the leukocyte telomere(P<0.05);in the T2 DM group,the age and BMI was negatively correlated with the telomere length of the leukocyte(P<0.05);in the DAS group,BMI and the time of illness was negatively correlated with the length of the leukocyte telomere(P<0.05).Detection of telomerase activity in peripheral blood leukocytes and plasma from patients with diabetes mellitus,according to the results of trap-pcr and RT q PCR showed that telomerase activity has not been found in T1 DM group and T2 DM group and NC group in peripheral blood plasma.Telomerase activity was found in peripheral blood leukocytes.The telomerase activity in peripheral blood leukocytes of T1 DM and T2 DMpatients were significantly lower than that of NC group(P<0.05);and the telomerase activity of T1 DM patients was higher than that of T2 DM patients(P<0.05).Multiple linear regression analysis showed that in the T2 DM group,there was a significant correlation between the gender and smoking status and the telomerase activity of leukocytes(P<0.05).So the telomerase activity of leukocytes in male and smoking patients was stronger than that of female and non-smokers.Conclusions: The telomere length of peripheral blood leukocytes from patients with diabetes was significantly shorter than that of normal people,and the length of the telomere of T1 DM patients was significantly shorter than that of T2DM;in contrast to type 2 diabetes,the telomere length of DAS patients was significantly shorter than that of T2 DM.The age,time of illness,and the BMI value of the patients were closely related to the shortening of telomere length.No telomerase activity was detected in the plasma of peripheral blood of patients with T1 DM and T2 DM.The telomerase activity in peripheral blood leukocytes of patients with T1 DM and T2 DM was significantly higher than that in normal controls.And the telomerase activity in T1 DM patients was higher than that in patients with T2 DM.In the T2 DM group,there was a significant correlation between gender,smoking status and the telomerase activity.Diabetes has a significant effect on the telomere length and telomerase activity in peripheral blood white blood cells.The examination of telomeres and telomerase may provide a reasonable basis for the early detection and early treatment.