Influence of different culture conditions on the phenotype,proliferation and cytoskeleton of VSMCs from rats / 解剖学报

ABSTRACT

ObjectiveTo study the influence of different culture conditions in vitro on phenotype, proliferation and cytoskeletal proteins expression of vascular smooth muscle cells(VSMCs). Methods The cultured VSMCs from rat aorta were divided into six groups P2 control,P2 starvation,P4 control,P4 starvation,P6 control and P6 starvation. The proliferating cells were labeled by 5-bromodeoxyuridine (5-BrdU); The mRNA expression of smooth muscle 22 alpha (SM22α) was detected by reverse transcription-polymerase chain reaction method (RT-PCR); The cytoskeletal proteins including SMα-actin,β-Tubulin and Desmin were observed through immunohistochemical staining. Results With the increase of cell passage, cytoskeletal proteins expression of VSMCs decreased,cellular organs increased and secretory vesicles were abundant; in serum-free cultured cells mitochondria increased and electron density enhanced in cytoplasm of VSMCs.On the contrary the expression of SMα-actin decreased, and the expression of SMα-actin increased. The expression of β-Tublin and Desmin decreased more obviously, and at 6 passages failed to express. Conclusion The conditioned medium and serum-free had the different effects on the phenotype,proliferation and cytoskeleton of VSMCs in different passage, and there was internal relationship among them. The internal relationship played an important role in the maintaining of cell morphology, contractile function and vascular remodeling. The disappearance of expression of β-Tubulin and desmin might have important biological significance.