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目的 探讨肿瘤坏死因子α(TNF-α)及其第 30 8位 G→ A变异、循环瘦素与 2型糖尿病 (2型 DM)胰岛素抵抗的相关情况。 方法 (1)测定全部受试对象的体质量指数 (BMI)及空腹静脉血糖 (FPG)、血清甘油三酯 (TG)、总胆固醇 (TC)等指标。 (2 )采用放射免疫的方法检测 2型 DM患者 70例和健康对照组 6 0例空腹血清胰岛素 (FINS)、TNF-α及瘦素水平。用HOMA胰岛素抵抗指数 (HOMA- IR=FPG× FINS/ 2 2 .5 )评价 2型 DM患者的胰岛素抵抗情况。 (3)采用饱和氯化钠法提取所有受试者外周血基因组 DNA。 (4 )以聚合酶链反应(PCR)扩增 TNF- α基因第 30 8位特异性片段 ,扩增产物用限制性内切酶 N co 酶切 ,酶切产物用 12 %聚丙烯酸胺凝胶电泳 ,于紫外灯下确定基因型。 (5 )以 HOMA- IR为因变量 ,以年龄、BMI、TG、TC、HDL、TNF- α基因型、血清 TNF- α及瘦素水平为自变量进行多元逐步回归分析 ,探讨胰岛素抵抗的影响因素。以瘦素为因变量 ,以年龄、BMI、TG、TC、HDL、FPG、FINS、TNF-α及 TNF-α基因型为自变量进行多元逐步回归分析 ,探讨瘦素的影响因素。 结果 (1) 2型 DM组与正常对照组 FPG (mmol/ L )分别为 11.0 3± 4 .89,5 .2 0± 5 .14 (P <0 .0 1) ,TNF-α(ng/ m L )为 1.2 6± 0 .2 8,1.15± 0 .2 4 (P
Objective To investigate the relationship between tumor necrosis factor-α (TNF-α) and G-A mutation at the 308th position, and the relationship between circulating leptin and type 2 diabetes mellitus (type 2 DM). Methods (1) The body mass index (BMI), fasting venous blood glucose (FPG), serum triglyceride (TG) and total cholesterol (TC) were measured in all the subjects. (2) The levels of fasting serum insulin (FINS), TNF-α and leptin in 70 DM patients and 60 healthy people were detected by radioimmunoassay. Insulin resistance in patients with type 2 DM was assessed using the HOMA insulin resistance index (HOMA-IR = FPG × FINS / 2 2 .5). (3) Peripheral blood genomic DNA was extracted from all subjects by saturated sodium chloride method. (4) Amplification of the 30 8th specific fragment of TNF-α gene by polymerase chain reaction (PCR), amplification products were digested with restriction endonuclease N co, the digested products were treated with 12% polyacrylamide gel Electrophoresis, in the UV light to determine the genotype. (5) By using HOMA-IR as the dependent variable, multiple stepwise regression analysis was conducted with age, BMI, TG, TC, HDL, TNF-α genotype, serum TNF- α and leptin as independent variables to explore the impact of insulin resistance factor. Using leptin as a dependent variable, multiple stepwise regression analysis of age, BMI, TG, TC, HDL, FPG, FINS, TNF-α and TNF-α genotypes were performed to explore the influencing factors of leptin. Results (1) The FPG (mmol / L) in type 2 DM group and normal control group were 11.0 3 ± 4.89, 5 .2 ± 5 .14, respectively (P < m L) was 1.2 6 ± 0.28, 8.15 ± 0.24 (P