bromochloroacetic-acid and Arteriosclerosis

Phenotypic modulation of smooth muscle cells (SMC) is a key event during the development of atherosclerotic and restenotic lesions. During this process, the composition of the cytoskeleton is substantially altered, with changes predominantly in actin expression reflecting a shift from smooth muscle alpha-actin to the non-muscle beta-isoform. We now demonstrate that yet another actin isoform, cardiac alpha-actin, is synthesized, de novo, in SMC of various atherosclerotic lesions. Using a highly specific monoclonal antibody against cardiac alpha-actin, we analyzed and compared the accumulation of this actin isoform in diverse SMC by immunofluorescence microscopy and immunoblotting. As expected, cardiac alpha-actin was present in human myocardium but not in healthy SMC of adult aorta, coronary arteries, trabeculae of the spleen, colon, stomach or skeletal muscle. Interestingly, the presence of cardiac alpha-actin was detected in umbilical cord vessels, human myometrium, in atherosclerotic coronary lesions and atherosclerotic lesions from peripheral vascular disease. The distribution of cardiac alpha-actin often paralleled that of cytokeratins 8 and 18, intermediate filament proteins typically found in dedifferentiated SMC. Taken together, the data presented here illustrate the expression of cardiac alpha-actin to be limited to either fetal vessels or those vessels or tissue having suffered damage or atrophy, outside its 'native' environment in the heart. The demonstration of cardiac alpha-actin in SMC of umbilical cord vessels and in atherosclerotic lesions but not in apparently healthy vessels supports the notion that SMC in atherosclerotic lesions exhibit a dedifferentiated phenotype.

Topics: Actins; Aorta; Arteriosclerosis; Coronary Vessels; Female; Humans; Keratins; Microscopy, Fluorescence; Muscle, Smooth, Vascular; Myocardium; Pregnancy; Protein Isoforms; Umbilical Veins

In the arterial wall, smooth muscle cells (SMC) normally exist in a quiescent, differentiated state, representing the contractile phenotype. During the development of atherosclerosis SMC change towards the synthetic phenotype going along with proliferation, chemotactic response and increased monocyte binding. Expression of monocyte chemoattractant protein-1 (MCP-1), a potent chemoattractant for monocytes, has been shown to be among the earliest events in atherogenesis. We investigated the effect of MCP-1 on differentiated and dedifferentiated SMC. Differentiation of SMC was induced using Matrigel as a matrix for cultivation. MCP-1 was expressed in SMC by means of a recombinant adenovirus. Expression of MCP-1 led to dedifferentiation of SMC as demonstrated by induction of cytokeratin 18, a marker for the synthetic phenotype. Concurrently, migration was only detectable in MCP-1 expressing cells, whereas SMC infected with a control virus, coding for the nuclear-targeted lacZ gene showed no migration. The expression of intercellular adhesion molecule-1 (ICAM-1) could be demonstrated in synthetic SMC and was induced after infection of differentiated cells with recombinant adenovirus, coding for MCP-1 (AdMCP-1). Expression of ICAM-1 was associated with a tenfold higher monocyte binding compared to lacZ infected cells. Our data suggest that MCP-1 plays an important role for SMC in the functional switch from the contractile to the synthetic phenotype in the course of atherogenesis.

Topics: Adenoviridae; Arteriosclerosis; Base Sequence; Cell Adhesion; Cell Culture Techniques; Cell Differentiation; Chemokine CCL2; DNA, Complementary; Gene Expression Regulation; Genetic Markers; Humans; Immunoblotting; Immunohistochemistry; Intercellular Adhesion Molecule-1; Keratins; Molecular Sequence Data; Monocytes; Muscle, Smooth, Vascular; Phenotype; Polymerase Chain Reaction; Sensitivity and Specificity

During development of atherosclerotic lesions, vascular smooth muscle cells (SMCs) undergo changes both phenotypically and in their cytoskeleton composition. An expression of cytokeratins 8 and 18 in SMCs in plaques of the human superficial femoral artery and of cytokeratin 8 in lesions of the aorta was recently described. Since cytokeratins are epithelial markers generally not found in normal adult vascular SMCs, we performed a detailed immunofluorescence microscopy study using a large panel of antibodies against the various cytokeratin polypeptides and other elements of the cytoskeleton. We included lesions of carotid, common and superficial femoral, iliac, and popliteal arteries; the abdominal aorta; and saphenous vein bypass grafts, as well as primary, restenotic, and transplantation-associated lesions of coronary arteries (n = 33). Cytokeratins 8 and 18 were present in myointimal cells of all pathological specimens. Colocalization with smooth muscle alpha-actin identified most cytokeratin-positive cells as SMCs. Only very few cells cosynthesized cytokeratin and desmin, whereas the majority of cytokeratin-positive cells were vimentin-positive. This pattern of cytoskeletal protein synthesis is similar to that found in some fetal and/or neonatal SMCs. These findings suggest that the synthesis of cytokeratins in a subset of SMCs of atherosclerotic lesions is a common phenomenon in coronary artery and peripheral vascular disease as well as graft disease and transplantation-associated arteriosclerosis and that the state of these SMCs is of a "dedifferentiated" fetal type.

Topics: Aorta; Arteriosclerosis; Coronary Artery Bypass; Coronary Disease; Coronary Vessels; Femoral Artery; Fluorescent Antibody Technique; Humans; Keratins; Muscle, Smooth, Vascular; Peripheral Vascular Diseases; Saphenous Vein; Vascular Diseases

An immunofluorescence method was used to study the expression of cytokeratin 8 in human aortic smooth muscle cells (SMCs) during prenatal development and in atherosclerotic plaques. Aortic SMCs from a 10-wk-old fetus contained cytokeratin 8 in additional to vimentin and a small amount of desmin, whereas, in the cells from a 25-wk-old fetus, cytokeratin 8 was not detected. Cytokeratin 8 was found in the SMCs from intimal thickenings, fatty streaks, and atherosclerotic fibrous plaques. Clusters of cytokeratin 8-positive cells were more abundant in rather advanced lesions (fibrous plaques) that contained at least some amount of lipid. Expression of cytokeratin 8 in the cells of human atherosclerotic lesions probably reflects general rearrangement of gene expression in the intimal cells.

Topics: Adult; Aged; Aged, 80 and over; Aorta; Arteriosclerosis; Child, Preschool; Cholesterol Esters; Embryonic and Fetal Development; Endothelium, Vascular; Female; Fetus; Fluorescent Antibody Technique; Humans; Keratins; Middle Aged; Muscle, Smooth, Vascular; Staining and Labeling

Topics: Aged; Arterial Occlusive Diseases; Arteriosclerosis; Cells, Cultured; Female; Humans; Keratins; Male; Middle Aged; Muscle, Smooth, Vascular; Recurrence

Using immunofluorescence microscopy we show that cells expressing cytokeratins 8 and 18 are frequently enriched in human vascular wall tissue pathologically altered by the appearance of intimal thickenings and atherosclerotic plaques. These cytokeratins occur in cells which also synthesize IFs containing vimentin and/or desmin, and a considerable proportion of the cytokeratin-positive cells has been identified as smooth muscle cells by colocalization of desmin and/or smooth muscle type alpha-actin. The presence of extremely low concentrations of these cytokeratins in such vascular tissues has been confirmed by gel electrophoresis with immunoblotting as well as by Northern blot hybridization using specific cytokeratin cRNA probes. The results are discussed in relation to the recent demonstration that low-level synthesis of cytokeratins 8 and 18 occurs in other muscular tissues and to the specific proliferative activity of these cells.

Topics: Arteriosclerosis; Cells, Cultured; Humans; Immunohistochemistry; Keratins; Muscle, Smooth, Vascular

Topics: Arteriosclerosis; Hematoxylin; Humans; Keratins; Proteins; Skin; Staining and Labeling

The denture-bearing mucosa of aged maxillary and mandibular ridges show morphologic changes that are irreversible. These changes limit the extensibility and the rebound capacity of the denture-bearing area.

Topics: Adolescent; Aged; Aging; Alveolar Process; Arteriosclerosis; Collagen; Connective Tissue; Female; Humans; Keratins; Male; Mouth Mucosa; Mouth, Edentulous; Osteoporosis