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Nan Fang Yi Ke Da Xue Xue Bao ; 35(7): 971-6, 2015 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-26198945

RESUMO

OBJECTIVE: To study the effect of platelet-derived growth factor-BB (PDGFBBB) on rat corpus cavernosum smooth muscle (CCSM) cell proliferation, migration and phenotypic modulation and explore the underlying mechanisms. METHODS: Wistar rat CCSM cells were obtained through a modified tissue culture method and identified by immunofluorescence assay. The effect of PDGFBB on the proliferation of CCSM cells was investigated using a CCK-8 kit and the optimum PDGFBB concentration for cell treatment was determined. CCSM cells were treated with vehicle or PDGF-BB at the optimum concentration, and the cell migration was examined using scratch assay; the mRNA expression of the transcription factor myocardin and the contractile phenotype markers αSMA and SMMHC in CCSM cells were determined by qRT-PCR at 24 h and 48 h. The protein expression of myocardin in CCSM cells incubated with PDGFBB for 0, 24 and 48 h was examined by Western blotting. RESULT: In CCSM cell culture, 96.5%and 96% of the cells were positive for αSMA and smoothelin, respectively. PDGFBB at different concentrations markedly promoted the proliferation of CCSM cells; the optimum PDGFBB concentration for enhancing cell proliferation was 12.5 ng/mL, which induced the migration of CCSM cells and significantly reduced the mRNA expressions of myocardin, αSMA and SMMHC (P<0.01). Exposure to PDGFBB decreased the protein expression of myocardin as the exposure time extended (within 48 h). CONCLUSION: CCSM cells of a high purity can be obtained by the modified tissue culture method. PDGFBB can promote the proliferation and migration of CCSM cells and cause a phenotypic conversion from the contractile to the synthetic type possibly by down-regulating myocardin.


Assuntos
Miócitos de Músculo Liso/efeitos dos fármacos , Pênis/citologia , Proteínas Proto-Oncogênicas c-sis/farmacologia , Actinas/metabolismo , Animais , Becaplermina , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Regulação para Baixo , Masculino , Miócitos de Músculo Liso/citologia , Cadeias Pesadas de Miosina/metabolismo , Proteínas Nucleares/metabolismo , Fenótipo , RNA Mensageiro , Ratos , Ratos Wistar , Transativadores/metabolismo
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