Journal of Southern Medical University ›› 2015, Vol. 35 ›› Issue (07): 971-.
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Abstract: objective To study the effect of platelet-derived growth factor-BB (PDGFBBB) on rat corpus cavernosum smoothmuscle (CCSM) cell proliferation, migration and phenotypic modulation and explore the underlying mechanisms. MethodsWistar 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 PDGFBBconcentration for cell treatment was determined. CCSM cells were treated with vehicle or PDGF-BB at the optimumconcentration, and the cell migration was examined using scratch assay; the mRNA expression of the transcription factormyocardin and the contractile phenotype markers α-SMA and SMMHC in CCSM cells were determined by qRT-PCR at 24 hand 48 h. The protein expression of myocardin in CCSM cells incubated with PDGFBB for 0, 24 and 48 h was examined byWestern blotting. Result In CCSM cell culture, 96.5% and 96% of the cells were positive for α-SMA and smoothelin,respectively. PDGFBB at different concentrations markedly promoted the proliferation of CCSM cells; the optimum PDGFBBconcentration for enhancing cell proliferation was 12.5 ng/mL, which induced the migration of CCSM cells and significantlyreduced the mRNA expressions of myocardin, α-SMA and SMMHC (P<0.01). Exposure to PDGFBB decreased the proteinexpression of myocardin as the exposure time extended (within 48 h). Conclusion CCSM cells of a high purity can be obtainedby the modified tissue culture method. PDGFBB can promote the proliferation and migration of CCSM cells and cause aphenotypic conversion from the contractile to the synthetic type possibly by down-regulating myocardin.
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https://www.j-smu.com/EN/Y2015/V35/I07/971