Roles of AMP-Activated Protein Kinase in Diabetes-Induced Retinal Inflammation
Roles of AMP-Activated Protein Kinase in Diabetes-Induced Retinal Inflammation
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DOI：
10.1167/iovs.11-8041
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发表时间：
2011-11-01
影响因子：
4.4
通讯作者：
Tsubota, Kazuo
中科院分区：
文献类型：
--
作者：
Kubota, Shunsuke;Ozawa, Yoko;Tsubota, Kazuo
目的。AMP激活的蛋白激酶(AMPK)是细胞能量状态的感受器。本研究的目的是利用已知的AMPK激动剂白藜芦醇和AICAR(5-氨基咪唑-4-甲酰胺核糖核苷)在糖尿病视网膜病变的小鼠模型中阐明AMPK的作用。方法：C57BL/6小鼠死亡前24小时口服白藜芦醇50 mg/kg或AICAR 100 mg/kg，连续7天。用Western印迹分析或酶联免疫吸附试验检测视网膜组织中磷酸化及总AMPK、磷酸化核因子(NF)-kappaBp65、细胞间黏附分子(ICAM)-1和血管内皮生长因子(VEGF)的表达。用脱乙酰酶荧光法测定视网膜sirtuin(SIRT)1活性。用刀豆蛋白A凝集素灌流标记技术检测白细胞与视网膜血管的粘附性。结果：糖尿病诱导的小鼠视网膜AMPK去磷酸化，白藜芦醇或AICAR显著逆转。白藜芦醇和AICAR均能显著逆转糖尿病视网膜SIRT1失活和核因子-kappaB磷酸化。糖尿病小鼠给予白藜芦醇治疗后，可显著降低糖尿病小鼠视网膜白细胞黏附、ICAM-1和VEGF的表达，提示糖尿病视网膜炎症源于AMPK信号通路的下调，导致SIRT1失活和核因子-kappaB激活。这些数据还表明，AMPK激动剂白藜芦醇可能被用作糖尿病视网膜病变的治疗剂。(投资眼科VS科学。2011年；52：9142-9148)doi：10.1167/iovs.11-8041
PURPOSE. AMP-activated protein kinase (AMPK) is a sensor of cellular energy status. The purpose of the present study was to elucidate the roles of AMPK in the pathogenesis of diabetic retinopathy using the known AMPK activators resveratrol and AICAR (5-aminoimidazole-4-carboxamide ribonucleoside) in a mouse model.METHODS. C57BL/6 mice with streptozotocin-induced diabetes were treated with resveratrol orally at 50 mg/kg for 7 days or with AICAR intraperitoneally at 100 mg/kg 24 hours before death. Retinal protein levels of phosphorylated and total AMPK, phosphorylated nuclear factor (NF)-kappa B p65, intercellular adhesion molecule (ICAM)-1, and vascular endothelial growth factor (VEGF) were evaluated by Western blot analysis or enzyme-linked immunosorbent assay. Retinal activity of sirtuin (SIRT) 1 was measured by deacetylase fluorometric assay. Leukocyte adhesion to the retinal vasculature was examined with a concanavalin A lectin perfusion-labeling technique.RESULTS. Induction of diabetes in mice led to retinal AMPK dephosphorylation, which was significantly reversed by either resveratrol or AICAR. Either resveratrol or AICAR significantly reversed SIRT1 deactivation and NF-kappa B phosphorylation, both of which were induced in the diabetic retina. Administration of resveratrol to diabetic mice significantly reduced diabetes-induced retinal leukocyte adhesion, together with retinal expression of ICAM-1 and VEGF.CONCLUSIONS. The present findings reveal that diabetes-induced retinal inflammation stems from downregulation of the AMPK pathway, leading subsequently to SIRT1 deactivation and NF-kappa B activation. The data also suggest the potential use of the AMPK activator resveratrol as a therapeutic agent for diabetic retinopathy. (Invest Ophthalmol Vis Sci. 2011;52:9142-9148) DOI: 10.1167/iovs.11-8041