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目的研究2型糖尿病大鼠氧化应激与骨骼肌细胞葡萄糖转运蛋白4转位间的相关性。方法雄性SD大鼠40只,按照体重随机分为模型组、对照组和正常组。采用左下腹注射链脲佐菌素(STZ)50 mg·kg~(-1)并结合高脂高糖饮食,制备2型糖尿病大鼠模型。对照组和正常组,注射相同剂量的柠檬酸缓冲液,对照组喂以高脂高糖饲料,正常组喂以普通饲料。3个月后,处死各组动物,心脏取血,分离血清,以正常组和对照组作为对照,检测模型组各项生化指标和氧化应激指标;用免疫印迹法检测骨骼肌细胞GLUT4的转位。结果正常组和模型组的空腹血糖分别为(5.59±1.04),(36.68±11.95)mmol·L~(-1)、三酰甘油分别为(0.86±0.54),(2.73±2.13)mmol·L~(-1)、总胆固醇分别为(1.65±0.29),(3.08±1.35)mmol·L~(-1)、空腹胰岛素分别为(18.09±3.41),(133.68±42.87)m U·L~(-1)、CAT分别为(24.25±2.17),(5.96±2.69)U·mg~(-1)protein、MDA分别为(6.12±1.25),(20.25±0.71)nmol·mg~(-1)protein、Mn-SOD分别为(40.96±1.24),(35.58±1.44)U·mg~(-1)protein、GSH-PX分别为(366.05±29.95),(297.65±10.39)U·mg~(-1)protein,模型组大鼠各项生化指标和氧化应激水平与正常组比较差异有统计学意义(P<0.01)。模型组、对照组和正常组大鼠骨骼肌细胞膜GLUT4表达水平分别为1.03±0.02,0.42±0.13,0.18±0.05,模型组和对照组大鼠骨骼肌细胞膜GLUT4表达水平与正常组比较差异有统计学意义(P<0.01)。模型组大鼠的骨骼肌细胞膜GLUT4含量与CAT之间的相关系数为0.815(P<0.01),与MDA之间的相关系数为-0.798(P<0.01),与Mn-SOD之间的相关系数为0.799(P<0.01),与GSH之间的相关系数为0.752(P<0.01)。结论氧化应激对2型糖尿病大鼠骨骼肌细胞GLUT4转位有抑制作用,并且胞膜GLUT4表达水平与氧化应激存在相关性。
Objective To investigate the correlation between oxidative stress and glucose transporter 4 translocation in type 2 diabetic rats. Methods Forty male SD rats were randomly divided into model group, control group and normal group according to body weight. The rat model of type 2 diabetes was established by injecting streptozotocin (STZ) 50 mg · kg -1 on the left lower abdomen and combining with high-fat and high-sugar diet. The control group and the normal group, the same dose of citrate buffer injection, the control group fed with high-fat, high-sugar diet, normal group fed with normal feed. After 3 months, the animals in each group were sacrificed, the heart blood was taken and the serum was separated. The normal group and the control group were used as controls to detect various biochemical indexes and oxidative stress indexes of the model group. Western blotting was used to detect the changes of GLUT4 Bit. Results The fasting blood glucose of the normal group and the model group were (5.59 ± 1.04) and (36.68 ± 11.95) mmol·L -1, respectively. The values of triglyceride were (0.86 ± 0.54) and (2.73 ± 2.13) mmol·L ~ (-1) and total cholesterol were (1.65 ± 0.29) and (3.08 ± 1.35) mmol·L -1, respectively, and fasting insulin were (18.09 ± 3.41) and (133.68 ± 42.87) m U · L ~ (-1) and CAT were (24.25 ± 2.17) and (5.96 ± 2.69) U · mg -1 protein, MDA were (6.12 ± 1.25) and (20.25 ± 0.71) nmol · mg -1 ) protein and Mn-SOD were (40.96 ± 1.24) and (35.58 ± 1.44) U · mg -1 protein, GSH-PX were (366.05 ± 29.95) and (297.65 ± 10.39) U · mg ~ -1) protein. The biochemical and oxidative stress levels in the model group were significantly different from those in the normal group (P <0.01). The expression of GLUT4 in skeletal muscle cell membrane of model group, control group and normal group were 1.03 ± 0.02, 0.42 ± 0.13 and 0.18 ± 0.05, respectively. The difference of GLUT4 expression in skeletal muscle cell membrane between model group and control group was statistically Significance (P <0.01). The correlation coefficient between the content of GLUT4 and CAT in the model group was 0.815 (P <0.01), the correlation coefficient with MDA was -0.798 (P <0.01), and the correlation coefficient with Mn-SOD Was 0.799 (P <0.01), and the correlation coefficient with GSH was 0.752 (P <0.01). Conclusion Oxidative stress can inhibit the translocation of GLUT4 in skeletal muscle cells of type 2 diabetic rats, and the expression level of GLUT4 in plasma membrane is correlated with oxidative stress.