糖尿病患者外周血白细胞端粒长度与端粒酶活性变化的研究

发布时间：2018-04-20 23:01

本文选题：糖尿病 + 端粒　；参考：《河北大学》2017年硕士论文

【摘要】：背景:糖尿病是一种以高血糖为病理特征的代谢性疾病,若体内长期维持高血糖状态会引起一系列的并发症,例如:肾脏、肝脏、心脏、血管、眼等的损伤,对人体健康产生非常严重的影响。糖尿病的发病率也呈逐年上升状态,并且,全球没有非常有效的医疗手段治愈此类疾病。糖尿病患病初始没有任何表征,当病人到医院就诊时,大部分胰岛细胞已经失去功能,只能依靠药物维持血糖平衡。糖尿病早期发现、早期治疗成为当今的一项研究热点,通过分子生物学手段对外周血白细胞端粒长度与端粒酶活性进行检测以探讨白细胞端粒长度变化、端粒酶活性变化与糖尿病的相关性。方法:提取30例1型糖尿病(T1DM)、60例2型糖尿病(T2DM)、40例2型伴动脉硬化糖尿病(DAS)患者外周血白细胞基因组DNA,利用实时荧光定量PCR的方法检测基因组DNA的端粒长度,以36B4基因作为对照,检测不同分型糖尿病间以及糖尿病患者与正常健康人间的端粒长度变化情况。从而探究糖尿病对人类外周血白细胞端粒长度的影响;利用多元线性回归分析探究影响端粒长度的具体因素。分别提取40例T1DM患者、80例T2DM患者的外周血白细胞与血浆,利用TRAP-PCR的方法检测白细胞与血浆中是否存在端粒酶活性;再利用荧光定量PCR的方法检测端粒酶的活性;从而探究糖尿病对外周血白细胞、血浆的端粒酶活性影响情况,利用多元线性回归分析探究影响端粒酶活性的主要因素。结果:建立了检测端粒长度与端粒酶活性的可靠办法。T1DM、T2DM、DAS患者外周血白细胞端粒长度均小于健康对照(NC)组(P0.05);在不同分型的糖尿病对比中T1DM组端粒长度短于T2DM组与DAS组,DAS组比T2DM组更短。通过多元线性回归分析得到:在T1DM组中,年龄与白细胞端粒长度呈负相关(P0.05);T2DM组中,年龄、BMI与白细胞端粒长度呈负相关(P0.05);DAS组中,BMI、患病时间与端粒长度呈负相关(P0.05)。在糖尿病患者外周血白细胞、血浆的端粒酶活性检测试验中,根据TRAP-PCR与RT-q PCR的结果显示,在T1DM组、T2DM组与NC组中的外周血血浆中并没有发现端粒酶活性。在外周血白细胞中均发现了端粒酶活性,T1DM患者的外周血白细胞端粒酶活性与T2DM患者均显著高于NC组的(P0.05);并且T1DM患者的端粒酶活性高于T2DM患者(P0.05)。多元线性回归分析显示:在T2DM组中,性别与吸烟状况与白细胞端粒酶活性有显著相关性(P0.05),得到男性与吸烟患者的白细胞端粒酶活性强于女性与不吸烟者。结论:糖尿病患者的外周血白细胞端粒长度明显短于正常人,并且T1DM患者的端粒长度显著短于T2DM;在2型糖尿病的对比中,DAS患者的端粒长度明显短于T2DM。患者的年龄、患病时间、BMI值与端粒长度的缩短有密切的关系。在T1DM与T2DM患者的外周血血浆中都没有发现端粒酶活性存在。T1DM与T2DM患者的外周血白细胞端粒酶活性显著高于正常人;并且,T1DM患者的白细胞端粒酶活性高于T2DM患者。在T2DM组中发现性别与吸烟状况与白细胞端粒酶活性有显著相关性。糖尿病对于外周血白细胞的端粒长度、端粒酶活性均有明显影响;对于二者的相关检查可能对于糖尿病的早发现、早治疗提供合理依据。
[Abstract]:Background: diabetes is a metabolic disease characterized by hyperglycemia. The long-term maintenance of hyperglycemia in the body causes a series of complications, such as kidney, liver, heart, blood vessels, and eye injuries, which have a very serious impact on human health. The incidence of glycuria is also rising year by year, and the world has not It is very effective to cure such diseases. There is no sign of diabetes at the beginning. When the patient goes to the hospital, most of the islet cells have lost their function and can only rely on drugs to maintain the blood sugar balance. Early diabetes has been found, and early treatment has become a hot spot of research, by means of molecular biological means. Telomere length and telomerase activity were detected to investigate the changes of telomere length and telomerase activity and the correlation between telomerase activity and diabetes. Methods: 30 cases of type 1 diabetes mellitus (T1DM), 60 cases of type 2 diabetes (T2DM), 40 cases of type 2 atherosclerosis diabetes (DAS) patients with peripheral blood leukocyte genome DNA were used, and real-time quantitative PCR was used. The telomere length of genomic DNA was detected and the 36B4 gene was used as the control to detect the change of telomere length between diabetic patients and diabetic patients and normal healthy people. The effect of diabetes on the telomere length of human peripheral blood white blood cells was explored, and the multielement linear regression analysis was used to explore the effect of telomere length. Body factors. The peripheral blood leukocytes and plasma were extracted from 40 patients with T1DM and 80 patients with T2DM respectively. The telomerase activity in white cells and plasma was detected by TRAP-PCR, and the activity of telomerase was detected by fluorescence quantitative PCR, and the telomerase activity in peripheral blood leukocytes and plasma was investigated. The main factors affecting telomerase activity were analyzed by multiple linear regression analysis. Results: a reliable method for detecting telomere length and telomerase activity was established. The telomere length of peripheral blood leukocytes in patients with T2DM and DAS was less than that of the healthy control group (NC) (P0.05), and the telomere length of group T1DM in different types of diabetes was shorter than that of group T2DM and D (T2DM and D). Group AS, group DAS was shorter than group T2DM. By multiple linear regression analysis, age was negatively correlated with telomere length in leukocyte (P0.05) in group T1DM; in group T2DM, age, BMI was negatively correlated with telomere length of white blood cells (P0.05); in group DAS, BMI, negative correlation with end grain length (P0.05). In diabetic patients, peripheral blood leukocytes, plasma In the test of telomerase activity, according to the results of TRAP-PCR and RT-q PCR, the telomerase activity was not found in the T1DM group, in the peripheral blood plasma of the T2DM group and the NC group. The telomerase activity was found in the peripheral blood leukocytes, and the activity of the peripheral leukocyte telomeric enzyme in the peripheral blood of the T1DM patients was significantly higher than that of the NC group (P0.05). The telomerase activity of T1DM patients was higher than that of T2DM patients (P0.05). Multiple linear regression analysis showed that in group T2DM, there was a significant correlation between sex and smoking status and leukocyte telomerase activity (P0.05). The telomerase activity of white blood cells in male and smoking patients was stronger than that of women and non smokers. Conclusion: the peripheral blood of diabetic patients is white. Telomere length was significantly shorter than that of normal people, and the telomere length of T1DM patients was significantly shorter than that of T2DM. In the contrast of type 2 diabetes, the telomere length of DAS patients was significantly shorter than the age of T2DM. patients, the time of the disease, the BMI value was closely related to the shortened telomere length. No telomerase was found in the peripheral blood plasma of the patients with T1DM and T2DM. Telomerase activity in peripheral blood leukocytes in patients with active.T1DM and T2DM was significantly higher than that of normal people; and the telomerase activity of leukocytes in T1DM patients was higher than that of T2DM patients. In T2DM group, there was a significant correlation between sex and smoking status and telomerase activity in leukocytes. Telomere length and telomerase activity in peripheral blood leukocytes were found in diabetic patients. There is a significant effect; the relevant examination of the two may provide a reasonable basis for early detection and early treatment of diabetes.

【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

【参考文献】