反义寡脱氧核苷酸对人阴茎海绵体平滑肌细胞PDE5的抑制作用  

The inhibitive effect of antisenes oligodeoxynucleotide on the phosphodiesterase type 5 in smooth muscle cells of human corpus cavernosum

作　　者：白文俊[1] 侯树坤[1] 王晓峰[1] 闫征[1] 何培英[1] 胡小朋[1] 管考鹏[1] 

机构地区：[1]北京大学人民医院泌尿外科,100044

出　　处：《中华泌尿外科杂志》2002年第11期656-658,共3页Chinese Journal of Urology

基　　金：国家自然科学基金 (3 0 15 0 0 0 2 )

摘　　要：目的　观察反义寡脱氧核苷酸对人阴茎海绵体平滑肌细胞 5型磷酸二酯酶 (PDE5 )的抑制作用 ,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　将PDE5基因反义寡脱氧核苷酸与DOTAP转染人阴茎海绵体平滑肌原代细胞 ,Western印迹法检测转染后不同时间 (1～ 4 8h)海绵体平滑肌细胞内PDE5的浓度变化 ,观察反义寡脱氧核苷酸对平滑肌细胞内PDE5的作用。　结果　转染后 1～ 6h ,反义组人阴茎海绵体平滑肌原代细胞内PDE5表达量较对照组显著降低 (P=0 .0 0 0～ 0 .0 14 ) ;转染后 12～ 4 8h ,三组细胞内PDE5的表达比较 ,差别无显著性意义 (P =0 .189～0 .96 2 )。　结论　PDE5基因反义寡脱氧核苷酸对人阴茎海绵体平滑肌细胞PDE5有抑制作用。Objective To investigate the inhibitive effect of antisenes oligodeoxynucleotide(ASON) on the phosphodiesterase type 5(PDE5) in smooth muscle cells of human corpus cavernosum,and provide experimental groundwork for the gene therapy of erectile dysfunction. Methods PDE5 gene ASON (containing exon 1) was transfected into the corpus cavernosum smooth muscle cells with the presence of liposome DOTAP.Another sense oligodeoxynucleotide(SON) and 1% of bovine serum were also transducted into the cells as controls.PDE5 was probed and measured by Western blot at 1,2,4,6,10,24 and 48 h after transfection. Results After transfection(1～6 h), PDE5 in human corpus cavernosum smooth muscle cells was significantly lower than it was in controls( P = 0.000 ～0.014). Conclusions The results indicate that PDE5 gene antisense oligodeoxynucleotide has inhibitive effect on PDE5 in smooth muscle cells of human corpus cavernosum in vitro,and the present study provides experimental groundwork for the gene therapy of erectile dysfunction.

关 键 词：反义寡脱氧核苷酸 阴茎 海绵体 平滑肌细胞 PDE5 抑制作用 

分 类 号：R697[医药卫生—泌尿科学]