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Abstract
Abstract
Dexlipotam and Formation of AGEs in RPE-cells
Akkoyun I., Lin J., Feng J., Jonas J. B., Hammes H. P.
Universitäts-Augenklinik, V. Med. Klinik, Universitätsklinikum Mannheim der Universität Heidelberg, Mannheim
Purpose: Dexlipotam has been designed as drug for the treatment of diabetic polyneuropathy and vascular dysfunction. To investigate the effect of dexlipotam on advanced glycation endproducts (AGEs) in human retinal pigment epithelium (RPE) cells regarding the possible role of oxidative stress in the pathogenesis in age-related macular degeneration (AMD).
Method: Cell cultures of the human RPE-K1034 cell line were incubated with various concentrations of dexlipotam (0.0, 1.0, 3.0, 10.0 mM) for 1 and 6 hrs, respectively, under baseline conditions (5 mM glucose), in a medium containing 5mM glucose + 30 mM H2O2 for 1 hr, and in a medium containing 25mM glucose. Westernblot analysis was performed to detect N-carboxymethyl-lysine (CML) as a marker of AGEs/glycoxidation.
Results: Both, under baseline conditions, and upon additional exposure to 30 mM H2O2 for 1 hr, a ~80 kD CML-modified protein band was present. Co-incubation with increasing concentrations of dexlipotam up to 10.0 mM did not alter this modification. Unexpectedly, this modification was not observed when cells were incubated in high glucose for 72 hrs. However, adding dexlipotam (1.0, 3.0, 10.0 mM) induced a stronger CML modification of the ~80 kD protein band. Longer exposure to dexlipotam (6 hrs, 1.0 and