反义核酸对人体及兔阴茎海绵体平滑肌细胞PDE5的抑制作用

To investigate the inhibitive effect of antisenes oligodeoxynucleotide(ASON) on the phosphodiesterase type 5 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 hrs after transfection. Two of cNMP were probed and measured by ELISA at 1,2,4,6,10,24 and 48 hrs after transfection. Results After transfection(1~6 h), PDE5 in human corpus cavernosum smooth muscle cells was significantly lower than its was in controls（p=0.000~0.014）. 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）. Conclusion The PDE5 gene antisense oligodeoxynucleotide had been showed to manifest inhibitive effect on the phosphodiesterase type 5 in smooth muscle cells of human corpus cavernosum in vitro, the PDE5 gene antisense oligodeoxynucleotide 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..

5型磷酸二酯酶(PDE5)是海绵体平滑肌细胞内第二信使cGMP的特异性水解酶，在阴茎勃起过讨衅鸸丶饔谩１究翁馔ü逋庠嘌艘蹙ズＣ嗵迤交∠赴徒⑿弁媚Ｐ停⒁阅孀疾《拘鳳DE5的抑制作用本研究有助于了解PDE5在阴茎勃起中的作用，并为阴茎勃起δ苷习幕蛑瘟铺峁├砺酆褪笛榛