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目的观察抑制视网膜上葡萄糖转运蛋白-1(glucose transporter-1,GLUT1)对糖尿病视网膜病变中视锥细胞的影响。方法 27只8周龄C57BL/6小鼠按随机数字表法分为正常对照组、糖尿病对照组和GLUT1小干扰核糖核酸(siRNA)治疗组。腹腔注射链脲佐菌素建立糖尿病模型后,GLUT1 siRNA治疗组予以玻璃体腔注射靶向GLUT1的siRNA,正常对照组和糖尿病对照组注射等量非靶向性siRNA,以上操作每2周重复注射1次,共注射9次。建模第18周3组小鼠行明适应视网膜电图检查视锥细胞功能,免疫荧光共定位法和免疫印迹法检查视网膜GLUT1的表达,测定比较视网膜含糖量,通过免疫荧光共定位法检查视锥细胞的密度及形态改变。结果与糖尿病对照组相比,GLUT1 siRNA治疗组小鼠视网膜GLUT1表达明显下调,较正常对照组下降68.51%(P<0.01)。尽管糖尿病对照组和GLUT1 siRNA治疗组视网膜组织含糖量均高于正常对照组,但GLUT1 siRNA治疗组小鼠视网膜含糖量比糖尿病对照组低42.67%,差异有统计学意义(P<0.01);糖尿病对照组和GLUT1 siRNA治疗组小鼠的明适应视网膜电图的a波及b波振幅均低于正常对照组,但GLUT1 siRNA治疗组较糖尿病对照组分别高47.59%和42.61%,差异有统计学意义(P<0.01);形态学检查发现糖尿病对照组较GLUT1 siRNA组视锥细胞排列稀疏,外节形态更为短小。结论 GLUT1 siRNA通过抑制GLUT1表达,限制转运葡萄糖进入视网膜,降低视网膜局部含糖量从而对光感受器视锥细胞产生保护作用。
Objective To observe the effect of inhibiting glucose transporter-1 (GLUT1) on the cones in diabetic retinopathy. Methods 27 8-week-old C57BL / 6 mice were divided into normal control group, diabetic control group and GLUT1 small interfering RNA (siRNA) treatment group by random number table. After intraperitoneal injection of streptozotocin to establish a diabetic model, GLUT1 siRNA treatment group was given intravitreal injection of siRNA targeting GLUT1, and normal control group and diabetic control group were injected with an equal amount of non-targeted siRNA. The above procedure was repeated every two weeks for 1 Times, a total of 9 times. At the 18th week of modeling, the mice in each group were exposed to the retinal electroretinogram to examine the function of the cones. Immunofluorescent co-localization and Western blotting were used to examine the expression of GLUT1 in the retina. The retina sugar content was measured and compared by immunofluorescence co-localization Cone density and morphological changes. Results Compared with the diabetic control group, the GLUT1 siRNA treatment group showed a significant decrease in the expression of GLUT1 in the retina, which was 68.51% lower than that in the normal control group (P <0.01). Although the diabetic group and GLUT1 siRNA treatment group retina tissue sugar content were higher than the normal control group, GLUT1 siRNA treatment group mice retina sugar content 42.67% lower than the diabetic control group, the difference was statistically significant (P <0.01) The amplitude of a-wave and b-wave of bright adapted ERG in diabetic control group and GLUT1 siRNA-treated mice were lower than those in normal control group, but GLUT1 siRNA treatment group was 47.59% and 42.61% higher than diabetic control group respectively (P <0.01). Morphological examination showed that the cones of diabetic control group were sparsely arranged and the morphology of outer segments was shorter than that of GLUT1 siRNA group. Conclusion GLUT1 siRNA can protect the photoreceptor cone cells by inhibiting the expression of GLUT1, limiting the transport of glucose into the retina and reducing the local sugar content in the retina.