| 摘要: |
| 目的 观察通过抑制视网膜上葡萄糖转运蛋白1(glucose transporter-1,GLUT1)表达而减少葡萄糖转运进入视网膜对糖尿病小鼠微血管病变的影响。方法 36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,每组12只。腹腔注射链脲佐菌素建立糖尿病模型后,GLUT1siRNA组予以玻璃体腔注射1 μL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。建模成功后第21周,对3组小鼠行免疫印迹法检查视网膜GLUT1的表达并测定比较视网膜的含糖量,通过测定视网膜ICAM-1和TNF-α的表达及检查视网膜血管白细胞黏滞和血-视网膜内屏障渗漏以比较微血管病变程度。结果 GLUT1siRNA组GLUT1在神经视网膜层的表达较正常对照组表达约下降77.00%,仅为糖尿病对照组的8.07%,差异有统计学意义(P<0.01)。尽管糖尿病对照组和GLUT1siRNA组视网膜组织含糖量均高于正常对照组,但GLUT1siRNA组小鼠视网膜含糖量为糖尿病对照组的50.05%,差异有统计学意义(P<0.01);糖尿病对照组和GLUT1siRNA组小鼠视网膜ICAM-1和TNF-α的表达均高于正常对照组(P<0.01),但这两个炎症因子在GLUT1siRNA组表达仅分别为糖尿病对照组的66.14%(P<0.05)和54.76%(P<0.01);同时我们发现糖尿病对照组较GLUT1siRNA组有更多的白细胞黏附在视网膜血管中,视网膜铺片发现糖尿病对照组较GLUT1siRNA组其荧光渗漏区域较多,且渗漏面积也较大。结论 GLUT1siRNA通过抑制GLUT1表达,限制葡萄糖转运进入视网膜,从而降低视网膜含糖量,减轻了糖尿病视网膜的炎症反应和血-视网膜内屏障渗漏,对糖尿病视网膜病变可产生缓解作用。 |
| 关键词: 葡萄糖转运蛋白1 小分子干扰RNA 糖尿病视网膜病变 |
| DOI：10.3724/SP.J.1008.2015.00147 |
| 投稿时间：2014-11-10修订日期：2014-12-03 |
| 基金项目:国家自然科学基金(81160123),江西省卫计委中医药科研计划(2013B016),江西省教育厅青年科学基金项目(GJJ13046). |
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| Negative regulation of glucose transport alleviates microvasculature pathological changes of retinopathy in diabetic mice |
| SHI Ke*,ZHANG Yue-zhi,XIE Lin,YOU Zhi-peng,ZHOU Yue,WANG Chang-yun |
(Department of Ophthalmology, the Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the effect of glucose transporter-1(GLUT1) suppression on microvasculature pathological changes in diabetic mice. Methods Thirty-six 8-week-old C57BL/6 mice were randomly divided into normal control, diabetic control and GLUT1siRNA treatment groups. Diabetic model was established by intraperitoneal injection of streptozotocin. GLUT1siRNA treatment group received intravitreal injection of siRNA-mediated GLUT1, and the other two groups received equal amount of non-specific siRNA. Twenty-one weeks after diabetic induction, immunoblotting was conducted to examine the expression of GLUT1 and two inflammation factors: ICAM-1 and TNF-α. We also calculated the retinal glucose concentration. Leukostasis assay and vascular leakage assay were utilized to compare the microvasculature pathological changes between the two groups. Results The expression of GLUT1 was significantly down-regulated in GLUT1siRNA treatment group compared with the other two groups (P<0.01), decreased by 77.00% compared with the normal control group and only being 8.07% of the diabetic control group. Though retinal glucose concentration of diabetic control and GLUT1siRNA treatment groups were higher than that of normal control, that in GLUT1siRNA treatment group was only 50.05% that of the diabetic control group (P<0.01). The expressions of ICAM-1 and TNF-α in GLUT1siRNA treatment and diabetic control were significantly higher than that in the normal control group, and their expressions in GLUT1siRNA treatment group were 66.14% (P<0.05)and 54.76% (P<0.01)those of the diabetic control group, respectively. Moreover, compared with GLUT1siRNA treatment group, diabetic control group had more adherent leukocytes in the retinal microvasculature and larger areas of leaked FITC-labeled albumin retina. Conclusion GLUT1siRNA can limit glucose transport into the retina by inhibiting GLUT1 expression, thus decrease retinal glucose concentration and alleviate microvasculature changes of diabetic retinopathy. |
| Key words: glucose transporter type 1 small interfering RNA diabetic retinopathy |
