目的:通过长期注射糖基化终末产物(AGEs),建立一种简单可行的模拟糖尿病血管病变的大鼠模型,并进一步研究profilin-1在糖尿病血管病变中的作用。
方法:Sprague-Dawley大鼠尾静脉注射AGEs-BSA(25mg/kg/天)0、20、30、40和60天。然后,主动脉的形态变化,心,评估肾脏和profilin-1的表达。在培养的内皮细胞中,shRNAprofilin-1用于阐明profilin-1在AGEs诱导的血管内皮损伤和炎症反应中的作用。
结果:主动脉,心,注射AGE的大鼠肾脏有明显的形态学改变。此外,主动脉血管重塑的指标明显增加,伴随着profilin-1在主动脉中的表达增加,心,和肾脏和肾脏基底膜上的多糖含量。此外,在注射AGEs的大鼠主动脉和与AGEs孵育的内皮细胞中,profilin-1蛋白水平显著上调.shRNAprofilin-1显着减弱profilin-1,AGEs受体(RAGE)的上调表达,与AGEs一起孵育的内皮细胞中的NF-κB,以及降低高水平的ICAM-1,IL-8,TNF-α,ROS,和AGEs诱导的细胞凋亡。
结论:外源性AGEs可以在体内一定程度上模拟糖尿病血管病变,并增加糖尿病并发症靶器官中profilin-1的表达。阻断profilin-1减弱血管病变和炎症反应,提示其在AGEs介导的代谢记忆中的关键作用。
To construct a simple and feasible rat model to mimic diabetic vasculopathy by chronic injection of advanced glycation end products (AGEs) and further determine the role of profilin-1 in vasculopathy in AGE-injection rats.
Sprague-Dawley rats were injected with AGEs-BSA (25 mg/kg/day) for 0, 20, 30, 40, and 60 days by caudal vein. Then, the morphological changes in the aorta, heart, and kidney and the expression of profilin-1 were assessed. In cultured endothelial cells, shRNA profilin-1 was used to clarify the role of profilin-1 in AGEs-induced vascular endothelial lesions and inflammatory reactions.
The aorta, heart, and kidney of the AGE-injection rats had obvious morphological changes. Also, the indicators of vascular remodeling in the aorta significantly increased, accompanied by the increased expression of profilin-1 in the aorta, heart, and kidney and polysaccharide content on the kidney basement membrane. In addition, the protein level of profilin-1 was markedly upregulated in the aorta of AGEs-injected rats and endothelial cells incubated with AGEs. shRNA profilin-1 markedly attenuated the upregulated expression of profilin-1, receptor for AGEs (RAGE), and NF-κB in endothelial cells incubated with AGEs, as well as reduced the high levels of ICAM-1, IL-8, TNF-α, ROS, and apoptosis induced by AGEs.
Exogenous AGEs can mimic diabetic vasculopathy in vivo to some extent and increase profilin-1 expression in the target organs of diabetic complications. Blockade of profilin-1 attenuates vascular lesions and inflammatory reactions, suggesting its critical role in the metabolic memory mediated by AGEs.