Pan Yun-Gao, Liu Ji-Hong, Zhan Ying, Wang Tao, Wan Zhi-Hua, Li Zhong-Yuan, Liu Yun
Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Zhonghua Nan Ke Xue. 2009 Aug;15(8):689-92.
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.
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.
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.
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.
探讨5型磷酸二酯酶(PDE5)小干扰RNA(siRNA)对人阴茎海绵体平滑肌细胞中环磷酸鸟苷(cGMP)的影响,为RNA干扰(RNAi)技术应用于勃起功能障碍的治疗提供实验室依据。
成功构建表达3对特异性短发夹RNA(shRNA)的重组腺病毒rAd5-shRNA-PDE5A3。将人阴茎海绵体平滑肌细胞分为实验组、阴性对照组和空白对照组,分别用rAd5-shRNA-PDE5A3、腺病毒rAd5-mock和磷酸盐缓冲液进行转染。转染后24、48和72小时采用放射免疫分析法检测cGMP浓度,观察rAd5-shRNA-PDE5A3对阴茎海绵体平滑肌细胞cGMP的影响。
rAd5-shRNA-PDE5A3组阴茎海绵体平滑肌细胞中的cGMP水平显著高于rAd5-mock对照组和空白对照组(P<0.05),在转染后72小时最为显著。
rAd5-shRNA-PDE5A3可显著提高人阴茎海绵体平滑肌细胞中的cGMP水平,增强对PDE5基因的抑制作用。