The inhibitive effect of siRNA on the phosphodiesterase type 5 in smooth muscle cells of human corpus cavernosum / 中华泌尿外科杂志

ABSTRACT

Objective To investigate the inhibitive effect of siRNA(small interfering RNA,siRNA) 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 (ED). Methods Small interfering RNAs targeting PDE5 gene were systhesized by using web design software provided by Ambion,there siRNAs and control siRNA were systhesized by Ambion. SiRNAs were transfected into smooth muscle cells of human corpus cavernosum by using siPORTTM Lipid reagent;down-regulation of PDE5 mRNA was detected by RT-PCR;the inhibitive effect of PDE5 was detected by Western Blotting. Results The results of RT-PCR indicated siRNA1、siRNA2 and siRNA3 made down-regulations of PDE5 mRNA expression in the transfected groups 58.2%、14.9% and 11.8%;the PDE5 expression decreased 70.5%、19.8% and 17.3%;however the expression did not have different in control siRNA and frank group. Conclusions The synthesized siRNAs in vitro were able to down-regulate the expression of PDE5.There were different capabilities of the specific siRNAs down-regulation.It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for ED gene therapy.