【作者】 何媛媛；

【导师】 李素梅；

【作者基本信息】 安徽医科大学 ， 内科学， 2008， 硕士

【摘要】 目的:通过比较糖尿病有大血管病变病人和糖尿病无大血管病变病人的外周血单核细胞中核因子κB (NF-κB)的表达水平,来探讨NF-κB在糖尿病大血管病变发生、发展中的作用。方法:本实验收集了正常对照组20例(A组),其中男性11例,女性9例,平均年龄为(54.30±7.39)岁;2型糖尿病无大血管病变组(B组)30例,其中男性17例,女性13例,平均年龄为(54.53±9.19)岁,病程(4.50±3.50)年;2型糖尿病有大血管病变组(C组)22例,其中男性12例,女性10例,平均年龄为(56.45±7.30岁),病程(6.21±3.88)年。留取清晨空腹肘静脉血5ml,分离出单核细胞,采用酶联免疫吸附法、流式细胞仪计数法检测单核细胞中NF-κB表达水平。结果:1.组间流式细胞仪测得NF-κB表达水平比较:与正常对照组【(3.667±0.376)%】比较,糖尿病无大血管病变组单核细胞中NF-κB表达水平【(10.198±0.467)%】显著增高,伴大血管病变者进一步明显升高【(15.270±0.820)%】,p<0.01;与不伴大血管病变者比较,伴大血管病变者单核细胞中NF-κB水平明显升高,p<0.01。2.组间ELISA法测得NF-κB表达水平比较:与正常对照组【(263.695±45.746)pg/ml】比较,糖尿病不伴大血管病变组NF-κB水平【(362.777±27.900)pg/ml】显著增高,伴大血管病变者进一步显著升高【(450.800±24.320)pg/ml】,p<0.01;与不伴大血管病变者比较,伴大血管病变者NF-κB水平显著增高,(p<0.01)。3.相关性分析:直线相关分析显示流式细胞仪和ELISA法检测单核细胞中NF-κB的表达水平呈显著正相关(r=0.793,p<0.01);在糖尿病无大血管病变和糖尿病合并大血管病变2组中,流式细胞仪检测法测得的NF-κB与空腹血糖(FBG)、糖化血红蛋白(HbA1c)和HOMA-IR指数呈显著正相关(r,p分别为0.736, p<0.01; 0.827, p<0.01; 0.843, p<0.01); ELISA法测得的NF-κB与与FBG、HbA1c和HOMA-IR指数呈显著正相关(r, p分别为0.526, p<0.01; 0.531, p<0.01; 0.76, p<0.01)。结论:糖尿病患者外周血单核细胞中核因子NF-KB表达的增加,并参与了糖尿病并大血管病变的发生发展过程。目的:通过比较短期胰岛素强化治疗前后的初发2型糖尿病病人外周血单核细胞中核因子-κB (NF-κB)的表达水平的变化,来探讨胰岛素的抗炎作用和对大血管的保护作用的机制。方法:初发2型糖尿病人20例,男性14例,女性6例,平均年龄为(51.25±5.71)岁,空腹血糖(FBG)为(11.18±1.73)mmol/L,餐后血糖(P2hBG)为(17.92±2.08)mmol/L,给予胰岛素强化治疗2周,根据血糖水平调整胰岛素用量,将空腹及睡前血糖控制在3.9～8.3mmol/L,三餐2h后血糖<10mmol/L,血糖达标时间为(3.40±1.76)d。强化治疗前和治疗后分别测FBG、P2hBG、血白细胞计数(WBC),中性粒细胞百分比(N%)和空腹胰岛素,治疗前后留取清晨空腹肘静脉血5ml,分离出单核细胞,采用酶联免疫吸附法(ELISA)、流式细胞仪计数法(FCM)检测血单核细胞中NF-κB表达水平,选择同期参加门诊体检的正常对照者20例,男11例,女9例,平均年龄(54.30±7.39)岁。结果:1与正常对照组比较用ELISA法检测的糖尿病组单核细胞中NF-κB蛋白表达水平【(352.11±20.04)pg/mlVS(263.70±45.75)pg/ml】和用FCM法检测的糖尿病组单核细胞中NF-κB蛋白表达水平【(15.53±2.49)%VS(3.67±1.68)%】比较显著增高,p<0.01。2与胰岛素强化治疗前比较胰岛素强化治疗2周后,①FBG、P2hBG【治疗前分别为(11.18±1.73)和(17.92±2.08)mmol/L;治疗后分别为(6.37±1.07)和(8.27±2.12)mmol/L】显著下降(p<0.01);②WBC、N%【治疗前分别为(7.39±1.56)×109/L和(64.61±5.45)%;治疗后分别为(5.52±1.12)×109/L和(56.23±4.71)%】明显降低(p<0.01);③用FCM法和ELISA法检测的单核细胞中NF-κB蛋白表达水平【治疗前分别为(15.53±2.49)%和(352.11±20.04)pg/ml;治疗后分别为(12.22±2.80)%和(331.75±19.20)pg/ml】明显降低,p<0.01。3相关性分析采用流式细胞仪和ELASA两种方法测定的单核细胞内NF-κB蛋白表达水平呈显著正相关(r=0.642,p<0.01);强化治疗后FCM法检测的单核细胞中NF-κB水平降低与FBG的降低与FBG、P2hBG和LgHOMA-IR的下降均无相关性(r分别为0.02,0.10,0.23, p﹥0.05)。强化治疗后ELISA法检测NF-κB水平降低与FBG、P2hBG和LgHOMA-IR的下降均无相关性(r分别为0.37,0.25,0.14, p﹥0.05)。结论: 2型糖尿病患者体内存在低度炎症,胰岛素除降糖作用外,胰岛素强化治疗可降低单核细胞NF-κB表达,减轻炎症反应进而发挥其抗动脉粥样硬化的作用。更多还原

【Abstract】 Objective: To investigate the relationship between NF kappa-B (NF-κB) and the development of diabetic macroangiopathy in patients with type 2 diabetes, we compare the expression level of Nuclear Factor-κB in mononuclear cell of type 2 diabetes patients with and without macroangiopathy.Methods: Total 20 normal controls(group A, 45%women, mean age, 54.30±7.39years) and 30 diabetic patients without macro-angiopathy (group B, 43%women, mean age, 54.53±9.19years, course, 4.50±3.50years) and 22 diabetic patients with(group C, 45%women, mean age, 56.45±7.30years, course, 6.21±3.88years) macro-angiopathy were recruited, mononuclear cell were separated from whole blood, NF kappa-B (NF-κB) expression level in mononuclear cells were detected by enzyme-linked immunosorbent assay(ELISA) and flow cytometry (FCM).Results: 1.The expression of NF-κB detected by FCM group comparison: Compared with the control group【(3.667±0.376)%】, the expression of NF-κB in diabetics without macro-angiopathy【(10.198±0.467)%】were significantly higher(p<0.01), the expression of NF-κB in diabetics with macro-angiopathy【(15.270±0.820)%】were further higher; compared with the diabetics without macro-angiopathy group, the expression of NF-κB in diabetics with macro-angiopathy were significantly higher(p<0.01).2.The expression of NF-κB detected by ELISA group comparison: Compared with the control group【(263.695±45.746)pg/ml】, the expression of NF-κB in diabetics without macro-angiopathy【(362.777±27.900)pg/ml】were significantly higher(p<0.01), the expression of NF-κB in diabetics with macro-angiopathy【(450.800±24.320)pg/ml】 were further higher; compared with the diabetics without macro-angiopathy group, the expression of NF-κB in diabetics with macro-angiopathy were significantly higher(p<0.01).3.Correlation analysis: Linear correlation analysis showed that the expression level of NF-κB detected by FCM had positive relationship with the expression level of NF-κB detected by ELISA in diabetics(r=0.793, p<0.01); the expression level of NF-κB detected by FCM had positive relationship with FBG, HbA1c and LgHoma-IR(r=0.736, p<0.01; r=0.827, p<0.01; r=0.843, p<0.01); the expression level of NF-κB detected by ELISA had positive relationship with FBG, HbA1c and LgHoma-IR(r=0.526, p<0.01; r=0.531, p<0.01; r=0.76, p<0.01).Conclusion: Increased level of NF-κB in peripheral blood mononuclear might be risk factor for peripheral macroangiopathy in patients with diabetes. Objective: To investigate the anti-inflammatory effects of insulin by observing the effect of short-term intensive insulin therapy on the expression level of Nuclear Factor-κB in peripheral blood mononuclear cell of newly-diagnosed type 2 diabetics.Method: 20 newly-diagnosed type 2 diabetic patients(30%women, mean age (51.25±5.71)years,fasting blood glucose(FBG)(11.18±1.73)mmol/L,postprandial blood glucose(P2hBG)(17.92±2.08)mmol/L) )were treated by intensive insulin therapy for 2 weeks, mononuclear cell were separated from whole blood, NF-κB expression level in mononuclear cells were detected by enzyme-linked immunosorbent assay(ELISA) and flow cytometry (FCM), FBG , P2hBG , white blood cell count(WBC), neutrocyte percents(N%) and fasting serum insulin(FINS) were measured pre-treatment and post- treatment. The other 20 healthy volunteers(45%women; mean age, 54.30±7.39years) served as control subjects.Results1 The expression levels of Nuclear Factor-κB detected by ELISA【(352.11±20.04)pg/mlVS(263.70±45.75)pg/ml】and FCM【(15.53±2.49)%VS (3.67±1.68)%】in mononuclear cell of type 2 diabetes patients were significantly higher than those of control subjects(p<0.01).2 After short-term intensive insulin therapy for 2 weeks,①FBG and P2hBG were significantly decreased, which were(11.18±1.73)mmol/L VS (6.37±1.07)mmol/L and(17.92±2.08)mmol/L VS(8.27±2.12)mmol/L, p<0.01;②WBC and N% were much lower than those of pre-treatment, which were(7.39±1.56)×109/L VS (5.52±1.12)×109/L and (64.61±5.45)% VS(56.23±4.71)%, p<0.01;③Compared to those of pre-treatment, the expression level of Nuclear Factor-κB detected by ELISA【(352.11±20.04)pg/ml VS (331.75±19.20)pg/ml】and FCM【(15.53±2.49)% VS (12.22±2.80)%】in mononuclear cell of type 2 diabetes patients were significantly decreased(p<0.01).3 Correlation analysis: Linear correlation analysis showed that The expression levels of Nuclear Factor-κB detected by ELISA had significant and positive relationship with .The expression levels of Nuclear Factor-κB detected by FCM (r=0.642,p<0.01) ; after 2 weeks of insulin treatment, the descent of Nuclear Factor-κB level detected by FCM had had no relationship with the descent of FBG(r=0.02,p>0.05), P2hBG(r=0.10,p>0.05) and LgHOMA-IR(r=0.23,p>0.05); after 2 weeks of insulin treatment, the descent of Nuclear Factor-κB detected by ELISA had no relationship with the descent of FBG(r=0.37,p>0.05), P2hBG(r=0.25,p>0.05) and LgHOMA-IR(r=0.14,p>0.05).Conclusion: There are existed low-degree of inflammation reaction in type 2 diabetics, Intensive Insulin Therapy can decrease the level of NF-κB in peripheral blood mononuclear cell and reduce the inflammatory reaction.更多还原