PDE5基因反义寡脱氧核苷酸对人阴茎海绵体平滑肌细胞cNMP的影响  

The Effects of Antisenes Oligodeoxynucleotide on the Cyclic Nucleotide Monophosphates in Smooth Muscle Cells of Human Corpus Cavernosum

作　　者：白文俊[1] 侯树坤[1] 王晓峰[1] 闫征[2] 何培英[2] 邓庆平[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 [2]北京大学人民医院中心实验室,北京100044

出　　处：《中华男科学杂志》2002年第2期88-91,共4页National Journal of Andrology

基　　金：国家自然科学基金资助 (3 0 15 0 0 0 2 )

摘　　要：目的 :观察PDE5基因反义寡脱氧核苷酸对人阴茎海绵体平滑肌细胞内cAMP和cGMP的影响 ,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 (含第 1外显子部分序列 )与脂质转染试剂DOTAP共同转染人阴茎海绵体平滑肌原代细胞 ,以酶联免疫法分别检测转染后不同时间 (1～ 4 8h)海绵体平滑肌细胞内cAMP和cGMP的浓度变化 ,观察反义寡脱氧核苷酸对平滑肌细胞内cNMP的影响。　结果 :转染后 ,反义实验组平滑肌原代细胞内cGMP水平 (1～ 6h)显著高于对照组 (P <0 .0 1)。　结论 :PDE5基因反义寡脱氧核苷酸可以增加人阴茎海绵体平滑肌细胞cGMP水平 ,本研究有助于了解PDE5基因与cNMP在阴茎勃起中的作用 ,并为阴茎勃起功能障碍的基因治疗提供理论和实验基础。Objectives: To investigate the effects of antisense oligodeoxynucleotide(ASON) on the cyclic nucleotide monophosphates(cNMP) 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. Two of cNMP, cAMP and cGMP, were probed and measured by ELISA at 1,2,4,6,10,24 and 48 h after transfection. Results: After transfection, the level of cGMP(1～6 h) in human corpus cavernosum smooth muscle cells was significantly higher than that in controls( P <0.01). Conclusions: The PDE5 gene ASON had been showed to manifest stimulative effect on the cGMP in smooth muscle cells of human corpus cavernosum in vitro , and it provides experimental groundwork for the gene therapy of erectile dysfunction.

关 键 词：反义寡脱氧核苷酸 细胞转染 海绵体平滑肌细胞 环磷酸核苷酸 

分 类 号：R698[医药卫生—泌尿科学]