Immunohistochemical Evidence for an Increased Oxidative Stress and Carbonyl Modification of Proteins in Diabetic Glomerular Lesions

Abstract

Abstract. Advanced glycation end products (AGE) include a variety of protein adducts whose accumulation has been implicated in tissue damage associated with diabetic nephropathy (DN). It was recently demonstrated that among AGE, glycoxidation products, whose formation is closely linked to oxidation, such as carboxymethyllysine (CML) and pentosidine, accumulate in expanded mesangial matrix and nodular lesions in DN, in colocalization with malondialdehyde-lysine (MDA-lysine), a lipoxidation product, whereas pyrraline, another AGE structure whose deposition is rather independent from oxidative stress, was not found within diabetic glomeruli. Because CML, pentosidine, and MDA-lysine are all formed under oxidative stress by carbonyl amine chemistry between protein amino group and carbonyl compounds, their colocalization suggests a local oxidative stress and increased protein carbonyl modification in diabetic glomerular lesions. To address this hypothesis, human renal tissues from patients with DN or IgA nephropathy were examined with specific antibodies to characterize most, if not all, carbonyl modifications of proteins by autoxidation products of carbohydrates, lipids, and amino acids: CML (derived from carbohydrates, lipids, and amino acid), pentosidine (derived from carbohydrates), MDA-lysine (derived from lipids), 4-hydroxynonenal-protein adduct (derived from lipids), and acrolein-protein adduct (derived from lipids and amino acid). All of the protein adducts were identified in expanded mesangial matrix and nodular lesions in DN. In IgA nephropathy, another primary glomerular disease leading to end-stage renal failure, despite positive staining for MDA-lysine and 4-hydroxynonenal-protein adduct in the expanded mesangial area, CML, pentosidine, and acrolein-protein adduct immunoreactivities were only faint in glomeruli. These data suggest a broad derangement in nonenzymatic biochemistry in diabetic glomerular lesions, and implicate an increased local oxidative stress and carbonyl modification of proteins in diabetic glomerular tissue damage (“carbonyl stress”).

Reactivity of hydrazinophthalazine drugs with the lipid peroxidation products acrolein and crotonaldehyde

“…Melting points were recorded on a Reichert hot stage apparatus. 1 H and 13 C spectra were measured using either a Varian Gemini-2000 spectrometer ( 1 H 300 MHz, 13 C 75 MHz) or a Varian INOVA spectrometer ( 1 H 600 MHz, 13 C 150 MHz). The multiplicities of signals are reported as being: s, singlet; d, doublet; t, triplet; q, quartet; multiplet, an unassignable multiplicity or overlapping signals; br, broadened signal.…”

“…Lipid peroxidation accompanies many debilitating health conditions including inflammatory disorders, 1 cardiovascular disease, 2,3 neurodegenerative conditions [4][5][6] and diabetes. 7 Such oxidative fragmentation of unsaturated lipids generates multiple reactive carbonyls including malondialdehyde, various 4-hydroxy-2-alkenals, and a range of 2-alkenals such as acrolein 1 and crotonaldehyde 2. 3,[8][9] Acrolein 1 is also a ubiquitous environmental pollutant due to its formation during the combustion of organic matter.…”

“…Acrolein-adducted proteins have been detected in the affected tissues of several chronic degenerative diseases known to involve oxidative stress, including Alzheimer's disease 12 and diabetic kidney disease. 13 Given this contribution of short-chain 2-alkenals to oxidative cell injury, our laboratory began a search for nucleophilic drugs that attenuate the toxicity of these substances. 14,15 The goal is to identify nucleophilic compounds that react with toxic carbonyls at a faster rate than do cell macromolecules, sparing the latter and ensuring carbonyls are excreted as drug-carbonyl derivatives.…”

Exclusive formation of imino[4 + 4]cycloaddition products with biologically relevant amines: plausible candidates for acrolein biomarkers and biofunctional modulators

“…1). Antibodies 11 to these conjugates are widely used for the immunochemical detection of various disease states, including arteriosclerosis, [11][12][13][14] Alzheimer's disease, 15,16 tumors, [17][18][19][20][21] diabetes, [22][23][24][25][26] autoimmune diseases, 27,28 high blood pressure, 29 and others. [30][31][32][33][34] Alternatively, acrolein can react with polyamines to produce FDP conjugates that modulate the cytotoxicity of acrolein.…”

Polyamine modification by acrolein exclusively produces 1,5-diazacyclooctanes: a previously unrecognized mechanism for acrolein-mediated oxidative stress

“…Acroleinamino conjugates involving, for example, lysine δ-amino groups, 3-formyl-3,4-dehydropiperidine (FDP), 7 or 3-methylpyridinium (MP) derivatives 10 have been described. Antibodies 11 to these conjugates are widely used for the immunochemical detection of various disease states, such as arteriosclerosis, [11][12][13][14] Alzheimer's disease, 15,16 tumors, [17][18][19][20][21] diabetes, [22][23][24][25][26] autoimmune Scheme 1 1,5-Diazacyclooctanes are previously unrecognized products of the reaction of polyamines with acrolein. Proposed reaction mechanism governing the OH-and amine-mediated "head-to-tail" imino [4 + 4] disease, 27,28 high blood pressure, 29 and so on.…”