血小板源性生长因子-BB对SD大鼠阴茎海绵体平滑肌细胞表型转化影响的初步研究

Platelet-derived growth factor-BB induces phenotypic transformation of corpus cavernosum smooth muscle cells in SD rats

目的:研究血小板源性生长因子-BB(PDGF-BB)对SD大鼠阴茎海绵体平滑肌细胞(CCSMC)表型转化的影响。方法:改良组织块法原代培养大鼠CCSMC,并进行细胞免疫荧光鉴定。实验分为空白对照组和不同浓度(5、10、20、40 ng/ml)PDGF-BB组,处理24 h;以及空白对照组和20 ng/ml PDGF-BB作用细胞不同时间(24、48、72 h)组,利用qRT-PCR检测各组细胞α平滑肌肌动蛋白(α-SMA)、平滑肌肌球蛋白重链(SMMHC)、碱性调宁蛋白和骨桥蛋白(OPN)mRNA的表达。结果:原代培养CCSMCα-SMA阳性细胞率达95%以上。与空白对照组比较,不同浓度PDGF-BB作用大鼠CCSMC后α-SMA、SMMHC、碱性调宁蛋白的mRNA表达显著减少(P均<0.05);OPN mRNA表达水平显著上调(P<0.05)。与空白对照组比较,随20 ng/ml PDGF-BB作用细胞时间的延长,α-SMA、SMMHC、碱性调宁蛋白的mRNA表达显著减少,OPN mRNA表达水平上调,差异均有统计学意义(P均<0.05)。结论:PDGF-BB可促使SD大鼠CCSMC表型从收缩型向合成型转化。

Objective： To evaluate the effect of the platelet-derived growth factor-BB（ PDGF-BB） on the phenotypic transformation of corpus cavernosum smooth muscle cells（ CCSMC） in SD rats. Methods： CCSMCs were primarily cultured in the modified tissue sticking medium and subjected to immunofluorescence assay. The cells were divided into a blank control and four PDGF-BB groups,the latter exposed to 5,10,20,and 40 ng / ml of PDGF-BB,respectively,for 24 hours,and the cells in the 20 ng / ml PDGFBB group treated for 24,48,and 72 hours. Then the relative expressions of α-SMA,SMMHC,calponin,and OPN mRNA were determined by real-time fluorescence quantitative RT-PCR（ qRT-PCR）. Results： The α-SMA positive rate of the CCSMCs was over95%. Compared with the blank control group,the expression levels of α-SMA,SMMHC,and calponin mRNA were significantly decreased（ P〈0. 05） while that of OPN mRNA remarkably increased（ P〈0. 05） in the PDGF-BB groups. The 20 ng / ml PDGF-BB group also showed significantly downregulated expressions of α-SMA,SMMHC,and calponin mRNA（ P〈0. 05） and upregulated expression of OPN mRNA（ P〈0. 05） at 24,48,and 72 hours. Conclusion： PDGF-BB can induce the transformation of the phenotype of CCSMCs in SD rats from the contractile to the synthetic type.