[Impact of adenovirus-mediated PDE5-shRNAs on the free calcium level in the smooth muscle cells of rat corpus cavernosum].
OBJECTIVE
To observe the influence of recombinant adenovirus-mediated PDE5-shRNAs on the free calcium level in rat penile smooth muscle cells and to explore the feasibility of gene therapy for erectile dysfunction (ED).


METHODS
Smooth muscle cells of the rat corpus cavernosum were transfected with constructed rAd-rPDE5-shRNAs and then dyed with the calcium fluorescent probe Fluo-3/AM at 24, 48 and 72 hours. The dynamic changes of the calcium fluorescence intensity were observed under the laser scanning confocal microscope (LSCM). The relative level of intracellular calcium was determined by fluorescence indexes.


RESULTS
The fluorescence indexes of calcium at 24, 48 and 72 hours were 829.3 +/- 7.8, 801.5 +/- 9.5 and 856.3 +/- 8.7 in the rAd-rPDES-shRNAs group, significantly lower than in the rAd-mock (1106.3 +/- 10.8, 1121.3 +/- 10.2 and 1058.5 +/- 12.1) and blank control group (1076.6 +/- 9.7, 1133.4 +/- 11.2 and 1104.3 +/- 10.5) (P < 0.05).


CONCLUSIONS
Adenovirus mediated shRNAs of the target PDE5 gene can significantly decrease the intracellular calcium level in the smooth muscle cells of the rat corpus cavernosum.
Pan
Yun-gao
YG
Department of Urology, Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Liu
Ji-hong
JH
Zhan
Ying
Y
Wang
Tao
T
Wan
Zhi-hua
ZH
Li
Zhong-yuan
ZY
Liu
Yun
Y
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5 / Q000378:metabolism*; D007172:Erectile Dysfunction / Q000628:therapy; D020868:Gene Silencing; D015316:Genetic Therapy; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D010413:Penis / Q000166:cytology; D034741:RNA, Small Interfering / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D034741:RNA, Small Interfering; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5; D002118:Calcium