钙通道阻滞剂维拉帕米对转化生长因子-β1诱导尿道瘢痕成纤维细胞α-平滑肌肌动蛋白和细胞外基质的作用  

The effect of verapamil on transforming growth factor-β1 induced alpha-smooth muscle actin and extracellular matrix of fibroblasts in cultured derived urethral scars

作　　者：许宁[1] 薛学义[1] 郭树平[1] 李晓东[1] 魏勇[1] 

机构地区：[1]福建医科大学附属第一医院泌尿外科,福州350005

出　　处：《中华实验外科杂志》2013年第9期1924-1926,共3页Chinese Journal of Experimental Surgery

基　　金：福建省自然科学基金资助项目（2012J01340）;福建医科大学教授发展基金资助项目（JS10017）

摘　　要：目的 探讨维拉帕米对转化生长因子-βl（TGF-βl）诱导尿道瘢痕成纤维细胞α-平滑肌肌动蛋白（α-SMA）和细胞外基质的作用及其机制.方法 分离培养尿道瘢痕中成纤维细胞.实验分为5组,培养液中分别加入TGF-β1（5μg/L）及维拉帕米（0、10、50、100 μmol/L）.逆转录-聚合酶链反应（RT-PCR）检测各组α-SMA mRNA、Smad7 mRNA变化；酶联免疫吸附试验（ELISA）法测定细胞外基质的表达.结果 随着不同浓度维拉帕米加入,各组α-SMA mRNA表达水平分别为0.497±0.085、1.460 ±0.062、1.070 ±0.066、0.780±0.060、0.293±0.067,α-SMA mRNA表达受抑制（P＜0.05）；各组Smad7 mRNA表达水平分别为0.477±0.065、0.223±0.055、0.680±0.053、0.833±0.032、1.077±0.060,表达逐渐增强（P＜0.05）.维拉帕米抑制细胞外基质表达,抑制效应呈剂量依赖性（P＜0.05）.结论 维拉帕米可能通过增强Smad7表达,抑制TGF-β1诱导的尿道瘢痕成纤维细胞α-SMA mRNA及细胞外基质合成.Objective To investigate the effect of verapamil on transforming growth factor-β1 （TGF-β1） induced alpha-smooth muscle actin （α-SMA） and extracellular matrix of fibroblasts in cultured derived urethral scars,and the molecular mechanisms.Methods Fibroblasts were isolated from urethral scar and cultured in vitro.Cells were divided into five groups,cultured by cell medium containing 5 μg/L TGF-β1 and different doses of verapamil （0,10,50 and 100 μmol/L）.After 72 hours incuation,the levels of α-SMA mRNA and Smad7 mRNA expression were assayed by semiquantitative real-time reverse transcriptase-polymerase chain reaction （RT-qPCR） technique.The productions of collagenI,collagen Ⅲ,fibronectin and larninin were examined by enzyme linked immunosorbent assay （ELISA）.Results TGF-β1 can significantly induce α-SMA mRNA expression.The α-SMA mRNA in each group were 0.497 ± 0.085,1.460 ± 0.062,1.070 ±0.066,0.780 ± 0.060,0.293 ± 0.067,respectively.With the increase of the verapamil concentration,the expression of α-SMA mRNA significantly was suppressed.The Smad7 mRNA in each group were 0.477 ± 0.065,0.223 ± 0.055,0.680 ± 0.053,0.833 ± 0.032,1.077 ± 0.060,respectively.Verapamil increased the expression of Smad7 mRNA in a dose-dependent manner （P ＜ 0.05）.Verapamil decreased expression of the extracellular matrix in a dose-dependent manner （P ＜ 0.05）.Conclusion Verapamil can inhibit the excessive synthesis of TGF-β1 induced α-SMA mRNA and extracellular matrix of fibroblasts in cultured derived urethral scars.The mechanisms may regulate the way of TGF-β/Smads,increasing the expression of Smad7.

关 键 词：尿道瘢痕 成纤维细胞 维拉帕米 转化生长因子 细胞外基质 

分 类 号：R699[医药卫生—泌尿科学]