[Impact of rAd5-shRNA- PDE5A3 on cGMP in the smooth muscle cells of human corpus cavernosum].
OBJECTIVE
To investigate the effect of phosphodiesterase type 5 (PDE5) small interfering RNA (siRNA) on cyclic guanosine monophosphatethe (cGMP) in the smooth muscle cells of human corpus cavernosum, and to provide laboratory evidence for the application of the RNA interference (RNAi) technique for the treatment of erectile dysfunction.


METHODS
The recombinant adenovirus rAd5-shRNA-PDE5A3 expressing three pairs of specific shRNA was constructed successfully. The smooth muscle cells of human corpus cavernosum were divided into an experimental, a negative control and a blank control group, and transfected respectively with rAd5-shRNA-PDE5A3, adenovirus rAd5-mock and phosphate buffered saline. The concentration of cGMP was measured by radioimmunoassay at 24, 48 and 72 hours after transfection, and the effect of rAd5-shRNA-PDE5A3 was detected on the cGMP in the smooth muscle cells of the corpus cavernosum.


RESULTS
The cGMP level in the smooth muscle cells of the corpus cavernosum was significantly higher in the rAd5-shRNA-PDE5A3 group than in the rAd5-mock control and blank control groups (P < 0.05), most significantly at 72 hours after transfection.


CONCLUSIONS
The rAd5-shRNA-PDE5A3 can obviously increase the cGMP level in the smooth muscle cells of human corpus cavernosum, and enhance the inhibition of the PDE5 gene.
Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Yun-Gao
YG
Pan
Ji-Hong
JH
Liu
Ying
Y
Zhan
Tao
T
Wang
Zhi-Hua
ZH
Wan
Zhong-Yuan
ZY
Li
Yun
Y
Liu
D000256:Adenoviridae; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism*; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5 / Q000235:genetics*; D007172:Erectile Dysfunction / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D010413:Penis / Q000166:cytology; D034741:RNA, Small Interfering
D034741:RNA, Small Interfering; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5; D006152:Cyclic GMP