title: Effects of lectins on calcification by vesicles isolated from aortas of cholesterol-fed rabbits.
abstract: Advanced vascular calcification in atherosclerosis weakens arterial walls, thereby imposing a serious rupturing effect. However, the mechanism of dystrophic calcification remains unknown. Although accumulating morphological and biochemical evidence reveals a role for calcifiable vesicles in plaque calcification, the mechanism of vesicle-mediated calcification has not been fully explored. To study whether vesicles' membrane components, such as carbohydrates, may have a role in vesicle-mediated calcification, the effect of sugar-binding lectins on calcification was investigated. Atherosclerosis was developed by feeding rabbits with a diet supplemented with 0.5% cholesterol and 2% peanut oil for 4 months. Calcifiable vesicles were then isolated from thoracic aortas by collagenase digestion. The histological examination of aortas with hematoxylin counter-staining indicated abnormal formation of large plaques enriched with macrophage-derived foam cells. Fourier transform spectroscopy revealed mild calcification in aortas indicating that advanced stages of heavy calcification have yet to be reached. However, vesicles isolated from the aortas were capable of calcification in the presence of physiological levels of Ca(2+), Pi, and ATP. Thus, at this stage of atherosclerosis, aortas may start to produce calcifiable vesicles, but at a level insufficient for substantial formation of mineral in aortas. The assessments by FT-IR analysis and Alizarin red staining indicated that concanavalin A (Con A) substantially increased mineral formation by isolated vesicles. Con A also exerted a marked stimulatory effect on (45)Ca and (32)Pi deposition in a dose-dependent fashion with a half-maximal effect at 6-10 microg/ml. Either alpha-methylmannoside or alpha-methylglucoside, but not mannitol, at 10 mM abolished the stimulation. Con A stimulation was abolished after Con A was removed from calcifying media, suggesting that covalent binding may not be involved in the effect. Galactosides appear to also be implicated in (45)Ca and (32)Pi deposition since Abrus precartorius agglutinin, which specifically binds galactosides, enhanced the deposition. Neither wheat-germ agglutinin that binds N-acetylglucoside nor N-acetylgalactoside-specific Helix pomatia agglutinin was effective, suggesting that the acetylated forms of carbohydrate moieties are either absent in vesicles or may not be involved in calcification. None of these lectins exerted an effect on ATPase. Thus, the effects of lectins appeared to be mediated through interactions with carbohydrate moieties of calcifiable vesicles. Whether stimulation of vesicle-calcification by lectins is of pathological significance in atherosclerotic calcification requires further investigation.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS 66160-7410, USA. hhsu@kumc.com
authors.forename: H H
authors.initials: HH
authors.lastname: Hsu
authors.forename: O
authors.initials: O
authors.lastname: Tawfik
authors.forename: F
authors.initials: F
authors.lastname: Sun
mesh_terms: D000818:Animals; D001011:Aorta / Q000737:chemistry / Q000187:drug effects* / Q000648:ultrastructure; D001013:Aorta, Thoracic / Q000187:drug effects; D001161:Arteriosclerosis / Q000209:etiology / Q000503:physiopathology; D002113:Calcification, Physiologic / Q000187:drug effects*; D002791:Cholesterol, Dietary; D017364:Collagenases; D003208:Concanavalin A / Q000494:pharmacology; D004036:Diet, Atherogenic; D037102:Lectins / Q000494:pharmacology*; D011817:Rabbits; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D002791:Cholesterol, Dietary; D037102:Lectins; D003208:Concanavalin A; D017364:Collagenases