人胰岛素样生长因子1基因在海绵体平滑肌细胞中的表达及定位

The expression and localization of human insulin-like growth factor-1 in the corpus carvernosum smooth muscle cells of rat

目的观察人胰岛素样生长因子1基因在海绵体平滑肌细胞中的表达及定位情况,为阴茎勃起功能障碍的基因治疗提供理论和实验依据。方法采用Lipofectamine2000包裹PCDB-hIGF-1质粒和PCDB空载体,转染原代培养的大鼠阴茎海绵体平滑肌细胞,转染后24h分别采用RT-PCR法检测hIGF-1基因mRNA表达,Westernblot法检测hIGF-1蛋白表达,荧光免疫细胞化学法检测hIGF-1细胞定位及蛋白表达。结果RT-PCR检测到hIGF-1mRNA表达,Westernblot和荧光免疫细胞化学法均检测到hIGF-1蛋白表达,荧光免疫细胞化学法可见hIGF-1蛋白定位于阴茎海绵体细胞胞质中。结论PCDB-hIGF-1成功转染入大鼠阴茎海绵体平滑肌细胞,大鼠阴茎海绵体平滑肌细胞可表达hIGF-1基因,蛋白定位于胞质。

Objective To observe the expression and localization of human insulin-like growth factor-1(hIGF-1) in the smooth muscle cells of rat and give a proof for the gene therapy to the erectile disfunction with insulin-like growth factor-1. Methods PCDB-hIGF-1 was transfected into the corpus carvernosum smooth muscle cells with the presence of lipofectamine 2000.PCDB without carrying any gene and 1% of bovine serum were also transducted into the cells as the controls. hIGF-1 and the protein were detected by RT-PCR...