Advanced glycation end‑products affect the cytoskeletal structure of rat glomerular endothelial cells via the Ras‑related C3 botulinum toxin substrate 1 signaling pathway

Lan,Lei; Han,Yongsheng; Ren,Wei; Jiang,Jielong; Wang,Peng; Hu,Zhao

doi:10.3892/mmr.2015.3317

Advanced glycation end‑products affect the cytoskeletal structure of rat glomerular endothelial cells via the Ras‑related C3 botulinum toxin substrate 1 signaling pathway

Authors:
- Lei Lan
- Yongsheng Han
- Wei Ren
- Jielong Jiang
- Peng Wang
- Zhao Hu
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Affiliations: Department of Nephrology, Affiliated Qilu Hospital, Shandong University, Jinan, Shandong 250012, P.R. China, Department of Emergency Internal Medicine, Affiliated Anhui Provincial Hospital, Anhui Medical University, Hefei, Anhui 230001, P.R. China, Department of Nephrology, Affiliated Anhui Provincial Hospital, Anhui Medical University, Hefei, Anhui 230001, P.R. China
Published online on: February 6, 2015 https://doi.org/10.3892/mmr.2015.3317
Pages: 4321-4326

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Abstract

The present study aimed to determine the molecular mechanisms leading to the production of advanced glycation end‑products (AGEs) and their effect on the morphology and function of rat glomerular capillary endothelial cells (GECs). Primary rat GECs were treated with AGE‑modified human serum albumin (AGE‑HSA) and divided into groups according to AGE concentration and treatment time. The structure and distribution of cytoskeletal protein F‑actin and the cortical actin binding protein, cortactin, were analyzed using immunofluorescence and confocal microscopy. As the Ras‑related C3 botulinum toxin substrate 1 (Rac1) signaling pathway was previously identified to be involved in mediating the contraction of endothelial actin‑myosin activity, Rac1 was examined subsequent to treatment of the cells with the Rac1 agonist 2'‑O‑methyladenosine‑3',5'‑cyclic monophosphate (O‑Me‑cAMP) for 1 h using a pull‑down assay. Cell permeability was determined by the leakage rate of a fluorescein isothiocyanate fluorescent marker protein. AGE‑HSA treatment resulted in alterations in the structure and distribution of F‑actin and cortactin in a dose‑ and time‑dependent manner, while no effect was observed with HSA alone. The effect of AGE on the cytoskeleton was inhibited by the addition of O‑Me‑cAMP. AGE‑HSA significantly reduced the level of Rac1 activity (P<0.05); however, no effect was observed on total protein levels. Furthermore, AGE‑HSA treatment led to a significant increase in the permeability of endothelial cells (P<0.01), which was inhibited by O‑Me‑cAMP (P<0.01). The Rac1 signaling pathway is thus suggested to serve an important function in mediating AGE‑induced alterations in GEC morphology and function.

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