Enhanced proliferation and migration and altered cytoskeletal proteins in early passage smooth muscle cells from young and old rat aortic explants

Smooth muscle cell (SMC) proliferation, migration, and cytoskeletal protein expression were studied in cultured cells obtained from the aortic explants of young (6-month) and old (30-month) Fischer 344XNB rats. Second-passage SMC were cultured on coverslips, and cytoskeletal fibers were examined by immunofluorescence microscopy using antibodies specific for smooth muscle myosin, α-smooth muscle actin, vimentin, desmin, and tubulin. The cytoskeletal fiber density was quantified as fluorescence intensity by confocal microscopy. The proliferation of SMC was analyzed from the growth curve of cells grown in culture from 0 to 14 days, and a Boyden chamber assay was used to quantify the SMC migration rate. The diameter of fresh SMC digested enzymatically from old rat aortae was 52.4% larger than that of the cells from young animals (20.0 ± 3 μm vs 13.1 ± 2 μm, P < 0.05). In SMC cultured from old animals, the intensities of smooth muscle myosin, α-smooth muscle actin, and vimentin decreased by 59.6, 41.2, and 54.8%, respectively; desmin and tubulin increased by 46.1 and 65.1% (all P < 0.001). Compared to SMC isolated from young rat aortae, the number of SMC cultured (second passage) from the old rat aorta was increased by 48.4, 27.2, and 26.9%, respectively, at Days 3, 7, and 14 in culture (P < 0.05, P < 0.01, and P < 0.001). The migration rate of SMC cultured from old rats was 59.3% higher than that of the cells obtained from young rats. These data show that alterations of the SMC cytoskeleton occur concomitantly with changes in SMC proliferation and migration rate during aging, suggesting that the age-associated changes in cytoskeletal proteins may play a role in remodeling of the aortic wall during aging.