Effects of a diet rich in advanced glycation end products in the rat remnant kidney model.

BACKGROUND: Food-derived advanced glycation end product (AGE)-analogues, the Maillard reaction products (MRPs), are formed during heat processing. Mainly low molecular weight MRPs are absorbed partially into the circulation and subsequently excreted in urine. In the presence of renal insufficiency, their removal is impaired, with a prolonged increase in plasma levels. Although bioactivity of orally absorbed MRPs has been shown in both experimental and human studies, its relevance in renal insufficiency still is unclear. METHODS: In the rat remnant-kidney model (five-sixth nephrectomy [5/6NX]), effects of an AGE-rich and an AGE-poor diet were investigated during a period of 6 weeks and compared with effects in sham-operated healthy (control [CTRL]) rats on renal function (serum creatinine level and proteinuria). In the AGE-rich diet, 25% wt/wt of cornstarch was replaced by bread crusts. RESULTS: Despite pair feeding, the AGE-rich diet resulted in a significant increase in body weight, including weight of the kidney, liver, and heart, in both the CTRL and experimental groups. The AGE-rich diet also enhanced proteinuria in CTRL rats by a factor of 2 and in 5/6NX rats by a factor of 8. Renal function (serum creatinine level and creatinine clearance) in healthy CTRLs did not change significantly. In the 5/6NX group, glomerular filtration rate (GFR) tended to even higher levels. CONCLUSION: Administration of an AGE-rich diet for 6 weeks does not impair GFR, but induces an increase in proteinuria, in particular, in the 5/6NX rats, indicating detrimental effects on the kidney.

RAGE-mediated MAPK activation by food-derived AGE and non-AGE products.

Investigating the cellular effects of food compounds formed by heat treatment during processing, we recently demonstrated the expression of the receptor for advanced glycation endproducts (RAGE) and the p44/42 MAP kinase activation by casein-N(epsilon )-(carboxymethyl)lysine (casein-CML), a food-derived AGE, in the intestinal cell line Caco-2. In this work, we report a Caco-2 p44/42 MAP kinase activation by bread crust and coffee extract. After identification, quantification, and synthesis of two key compounds formed in association with the process-induced heat impact applied to bread dough and coffee beans, those compounds, namely the AGE pronyl-glycine and the non-AGE N-methylpyridinium, were also demonstrated for the first time to activate the p44/42 MAP kinase through binding to RAGE in Caco-2 cells. Blocking of RAGE by an antagonistic antibody and expression of C-terminally truncated RAGE resulted in a reduced Caco-2- and HEK-293-MAP kinase activation. These findings unequivocally point to a RAGE-mediated activating effect of chemically defined food-derived, thermally generated products, both, AGEs and non-AGEs, on cellular signal transduction pathways involved in inflammatory response and cellular proliferation.

Structural and functional characterization of pronyl-lysine, a novel protein modification in bread crust melanoidins showing in vitro antioxidative and phase I/II enzyme modulating activity.

Application of an in vitro antioxidant assay to solvent fractions isolated from bread crust, bread crumb, and flour, respectively, revealed the highest antioxidative potential for the dark brown, ethanol solubles of the crust, whereas corresponding crumb and flour fractions showed only minor activities. To investigate whether these browning products may also act as antioxidants in biological systems, their modulating activity on detoxification enzymes was investigated as a functional parameter in intestinal Caco-2 cells. The bread crust and, in particular, the intensely brown, ethanolic crust fraction induced a significantly elevated glutathione S-transferase (GST) activity and a decreased phase I NADPH-cytochrome c reductase (CCR) activity compared to crumb-exposed cells. Antioxidant screening of Maillard-type model mixtures, followed by structure determination, revealed the pyrrolinone reductones 1 and 2 as the key antioxidants formed from the hexose-derived acetylformoin and N(alpha)-acetyl-L-lysine methyl ester or glycine methyl ester, chosen as model substances to mimic nonenzymatic browning reactions with the lysine side chain or the N terminus of proteins, respectively. Quantitation of protein-bound pyrrolinone reductonyl-lysine, abbreviated pronyl-lysine, revealed high amounts in the bread crust (62.2 mg/kg), low amounts in the crumb (8.0 mg/kg), and the absence of this compound in untreated flour. Exposing Caco-2 cells for 48 h to either synthetically pronylated albumin or purified pronyl-glycine (3) significantly increased phase II GST activity by 12 or 34%, respectively, thus demonstrating for the first time that "pronylated" proteins as part of bread crust melanoidins act as monofunctional inducers of GST, serving as a functional parameter of an antioxidant, chemopreventive activity in vitro.