Collagen as a model system to investigate the use of aspirin as an inhibitor of protein glycation and crosslinking.
Aspirin has been shown to be a powerful inhibitor of post-Amadori Maillard reactions, although the exact mechanism of this action remains unclear. We have used corneal and scleral collagen as a model system: (i) to assess how aspirin, either alone or in combination with sugars, affects the surface charge distribution along the collagen fibrils; (ii) to see how sugars and/or aspirin affect the swelling properties of the cornea; and (iii) to see if sugars and/or aspirin change the distribution of water within the corneal stroma as the tissue swells. Charge changes were detected by examining changes in the uptake of phosphotungstate ions as seen in the electron microscope. Swelling was measured by monitoring the uptake of water as a function of swelling time, and water distribution was determined by using synchrotron X-ray diffraction to follow changes in the interfibrillar Bragg spacing as the cornea swells. Aspirin has a marked effect on the positive staining pattern of scleral collagen. This is different to the changes in stain uptake produced by glycation. Incubation with both sugars and aspirin showed that the sugar binding takes precedence over the effects of aspirin which, in turn, suggests that protein acetylation by aspirin is unlikely under these circumstances. However, aspirin completely suppresses corneal swelling. Even when the aspirin is removed, swelling in distilled water is reduced, and this is accompanied by changes in the water distribution. The results suggest that water is more evenly distributed in aspirin-treated corneas that are subsequently swollen than in swollen glycated corneas. Fructation, glucation and ribation on their own have little effect on the uptake of water as the cornea swells. This suggests that any sugar-derived crosslinks formed at this stage do not limit swelling.
Hadley
J
J
The Open University, Oxford Research Unit, Boars Hill, Oxford, UK.
Malik
N
N
Meek
K
K
D000818:Animals; D001241:Aspirin / Q000494:pharmacology*; D002417:Cattle; D003094:Collagen / Q000737:chemistry / Q000187:drug effects*; D003315:Cornea / Q000737:chemistry / Q000187:drug effects*; D015715:Corneal Edema; D003432:Cross-Linking Reagents / Q000737:chemistry; D005632:Fructose / Q000494:pharmacology; D017127:Glycation End Products, Advanced; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D008854:Microscopy, Electron; D008875:Middle Aged; D012590:Sclera / Q000737:chemistry / Q000187:drug effects*; D014961:X-Ray Diffraction
D003432:Cross-Linking Reagents; D017127:Glycation End Products, Advanced; D005632:Fructose; D003094:Collagen; D001241:Aspirin