Prevention of haemoglobin glycation by acetylsalicylic acid (ASA): A new view on old mechanism

Abstract

Diabetic hyperglycemia provokes glycation of haemoglobin (Hb), an abundant protein in red blood cells (RBCs), by increasing its exposure to carbohydrates. Acetylsalicylic acid (ASA; Aspirin) is one of the first agents, which its antiglycation effect was witnessed. Although the precise molecular mechanism of action of ASA on protein glycation is not indisputably perceived, acetylation as its main molecular mechanism has been proposed. This report aims to unravel the meticulous mechanism of action of ASA by using two ASA analogues; benzoic acid (BA) and para-nitrobenzoic acid (NBA), despite their lack of acetyl group. In this regard, the inhibitory effect of these two chemicals in comparison with ASA on Hb fructation is reported. UV-visible spectroscopy, intrinsic advanced glycation end products (AGE) fluorescence spectroscopy, extrinsic thioflavin T (ThT) binding fluorescence spectroscopy, 2,4,6-trinitrobenzenesulfonic acid (TNBSA) assay, and single cell gel electrophoresis (SCGE) were used to explore the effects of BA and NBA in comparison with aforementioned chemicals in the context of protein glycation. In spite of the lack of acetyl substitution, NBA is reported as a novel agent with prominent inhibitory efficacy than ASA on the protein glycation. This fact brings up a possible new mechanism of action of ASA and reconsiders acetylation as the sole mechanism of inhibition of protein glycation.

Association of Novel Advanced Glycation End-Product (AGE10) with Complications of Diabetes as Measured by Enzyme-Linked Immunosorbent Assay

“…Except for the most known AGE—carboxymethyllysine (CML), commercially available immunoassays detect a mixture of various AGEs. Still, a body of evidence is gathering showing that individual AGEs have indeed potential as markers of diabetic complications, e.g., a methylglyoxal-derived hydroimidazolone-1 is proposed as an early marker of atherosclerosis in childhood diabetes [ 36 ], AGE4 (albumin modified by methylglyoxal) is an independent predictor of polyneuropathy in diabetes [ 35 ] and AGE1 (albumin modified by glucose) is significantly associated with lipid abnormalities in diabetes [ 37 ], justifying efforts put into developing epitope-specific immunoassays.…”

“…Moreover, aspirin prevents glycation of plasma proteins, including hemoglobin, but the molecular mechanism is not fully understood. Probably, aspirin acetylates free amino groups of a protein, although it is likely only one aspect of its anti-glycation activity [ 36 ]. An example of a glycation product, the formation of which is inhibited by aspirin, is pentosidine.…”

Identification of the Protein Glycation Sites in Human Myoglobin as Rapidly Induced by d-Ribose

“…For the investigation of protein glycations, the most in-depth study is on glycated hemoglobin (Hb) [ 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ]. In 2010–2011, the American Diabetes Association (ADA) and the World Health Organization (WHO) recommended HbA1c, an adduct of glucose and Hb, as a new standard for the diagnosis of diabetes [ 18 ].…”

“…In 2010–2011, the American Diabetes Association (ADA) and the World Health Organization (WHO) recommended HbA1c, an adduct of glucose and Hb, as a new standard for the diagnosis of diabetes [ 18 ]. Meanwhile, different carbohydrates and proteins in the human body may undergo protein glycation, leading to alterations of the protein structure and function [ 6 , 7 , 8 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ]. For example, glucose and ribose were found to undergo protein glycation with the Hb [ 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ].…”

“…Meanwhile, different carbohydrates and proteins in the human body may undergo protein glycation, leading to alterations of the protein structure and function [ 6 , 7 , 8 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ]. For example, glucose and ribose were found to undergo protein glycation with the Hb [ 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ]. The glycation sites of glucose in the Hb were identified previously, including in the major sites of β-V1, α-K16, and β-K66, and other Lys residues in both α and β subunits, whereas the structures of the glycation products were not reported [ 10 , 12 ].…”