Role of oxidative stress in advanced glycation end product-induced mesangial cell activation
Role of oxidative stress in advanced glycation end product-induced mesangial cell activation
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DOI：
10.1046/j.1523-1755.2002.00367.x
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发表时间：
2002-06-01
影响因子：
19.6
通讯作者：
Aperia, A
中科院分区：
文献类型：
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作者：
Lal, MA;Brismar, H;Aperia, A
背景晚期糖基化终产物（AGE）水平在年龄增长、肾衰竭和糖尿病患者中升高，这些分子的积累可能导致疾病进展。然而，AGE蛋白改变肾小球系膜细胞功能的机制尚不完全清楚。本研究评估了氧化应激在AGE依赖性肾小球系膜细胞信号转导中的作用。将原代培养的大鼠肾系膜细胞暴露于体外AGE-BSA和H-2 O-2。核因子-κ B（NF-κ B）和蛋白激酶C（PKC）亚型的激活进行了研究，使用共聚焦显微镜和蛋白质印迹。采用定量聚合酶链反应（PCR）测定转化生长因子-β 1（TGF-β 1）水平。通过补充或损害细胞抗氧化能力来评估氧化应激的参与。NF-κ B被AGE剂量依赖性地激活。PKC激活不参与这种反应，但PKC-β 1激活的分析显示AGE蛋白对这种亚型的刺激作用。TGF-β 1的转录被AGE刺激，并被PKC抑制剂阻止。H2 O2刺激对系膜细胞信号传导有类似的下游效应.抗氧化剂，维生素E和硝替卡朋，防止AGE依赖性NF-κ B激活和正常化PKC活性和相关的TGF-β 1转录。细胞内抗氧化剂谷胱甘肽的耗尽有效地降低了系膜细胞激活NF-κ B和PKC-β 1所需的AGE浓度。在谷胱甘肽耗竭的条件下，用次优剂量的AGE治疗，显示了对两个参数的协同作用。这些结果支持了氧化应激在AGE依赖性系膜细胞信号传导中的核心作用，并强调了ROS在决定细胞反应性中的重要性。
Background. Levels of advanced glycation end products (AGE) are elevated in individuals with advancing age, renal failure, and diabetes, and accumulation of these molecules may contribute to disease progression. The mechanism by which AGE proteins alter glomerular mesangial cell function, however, is not completely understood. The present study assessed the involvement of oxidative stress in AGE-dependent mesangial cell signaling events.Methods. Primary cultures of rat renal mesangial cells were exposed to in vitro AGE-BSA and H-2 O-2 . Nuclear factor-kappaB (NF-kappaB) and protein kinase C (PKC) isoform activation were studied using confocal microscopy and Western blotting. Quantitative polymerase chain reaction (PCR) was used to measure transforming growth factor-beta1 (TGF-beta1) levels. The involvement of oxidative stress was assessed by supplementing or compromising cellular antioxidant capacity.Results. NF-kappaB was dose-dependently activated by AGE. PKC activation was not involved in this response, but analysis of PKC-beta1 activation showed a stimulatory effect of AGE proteins on this isoform. Transcription of TGF-beta1 was stimulated by AGE and was prevented by PKC inhibition. Challenge with H-2 O-2 had similar downstream effects on mesangial cell signaling. Antioxidants, vitamin E and nitecapone, prevented AGE-dependent NF-kappaB activation and normalized PKC activity and associated TGF-beta1 transcription. Depletion of the intracellular antioxidant, glutathione, effectively lowered the AGE concentration needed for mesangial cell activation of NF-kappaB and PKC-beta1. Treatment with a suboptimal AGE dose, under glutathione-depleted conditions, revealed a synergistic effect on both parameters.Conclusion. The results support a central role for oxidative stress in AGE-dependent mesangial cell signaling and emphasize the importance of ROS in determining cell responsiveness.