To investigate effects of verapamil on primary cultured human urethral scar fibroblasts (USFs) and to provide basis for protecting the formation of urethra scar. Methods: The cell proliferation was evaluated with the cell counting kit (CCK)-8 method after USFs were incubated various verapamil concentrations (50, 100, 150, 200, or 250 μmol/L) or solvent for 12, 24, or 48 h. The protein level of matrix metalloproteinase (MMP) was evaluated with ELISA after cells were incubated with verapamil (100 μmol/L) or solvent (control cells) for 24 h. Results: The proliferation of USFs was obviously suppressed after verapamil treatment, which was in a dose-dependent and time-dependent manner. Meanwhile, the protein levels of MMP-2 and MMP-9 in the verapamil treatment group increased obviously compared with those of the control groups (P<0.05). Conclusion: Calcium channel blockers may prevent the excessive formation of urethra scar by inhibiting the proliferation of urethral scar fibroblasts and enhancing the activity of MMP.
目的:探索钙通道阻滞剂维拉帕米对尿道瘢痕成纤维细胞的作用,为临床应用钙通道阻滞剂防治尿道瘢痕形成提供实验依据。 方法:采用组织块培养法行人尿道瘢痕成纤维细胞原代培养,传至第4~8代用于实验研究。采用细胞计数试剂盒(cell counting kit-8,CCK-8)比色法测定在不同给药浓度下(50,100,150,200及250 μmol/L维拉帕米)干预不同时间(12,24 及48 h)后,尿道瘢痕成纤维细胞抑制率;采用酶联免疫吸附试验法测定实验组(100 μmol/L维拉帕米,24 h)与对照组的细胞上清液中基质金属蛋白酶(matrix metalloproteinase,MMP)-2和MMP-9的表达情况。 结果:维拉帕米对尿道瘢痕成纤维细胞具有抑制作用,呈现浓度与时间依赖性(P<0.05)。维拉帕米干预后细胞上清液MMP-2和MMP-9的蛋白水平明显高于对照组(P=0.01)。结论:钙通道阻滞剂可以抑制尿道瘢痕成纤维细胞的增殖,提升基质金属蛋白酶的活性,这可能对尿道瘢痕的形成具有防治作用。.