[腺病毒介导的磷酸二酯酶5短发夹RNA对大鼠阴茎海绵体平滑肌细胞游离钙水平的影响]

[Impact of adenovirus-mediated PDE5-shRNAs on the free calcium level in the smooth muscle cells of rat corpus cavernosum].

作者信息

Pan Yun-gao, Liu Ji-hong, Zhan Ying, Wang Tao, Wan Zhi-hua, Li Zhong-yuan, Liu Yun

机构信息

Department of Urology, Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.

出版信息

Zhonghua Nan Ke Xue. 2009 Oct;15(10):882-5.

PMID:20112734

Abstract

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).

CONCLUSION

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.

摘要

目的

观察重组腺病毒介导的PDE5 - shRNAs对大鼠阴茎平滑肌细胞游离钙水平的影响，探讨基因治疗勃起功能障碍（ED）的可行性。

方法

用构建好的rAd - rPDE5 - shRNAs转染大鼠海绵体平滑肌细胞，于转染后24、48和72小时用钙荧光探针Fluo - 3/AM进行染色。在激光扫描共聚焦显微镜（LSCM）下观察钙荧光强度的动态变化。通过荧光指标测定细胞内钙的相对水平。

结果

rAd - rPDES - shRNAs组在24、48和72小时的钙荧光指标分别为829.3±7.8、801.5±9.5和856.3±8.7，显著低于rAd - mock组（1106.3±10.8、1121.3±10.2和1058.5±12.1）及空白对照组（1076.6±9.7、1133.4±11.2和1104.3±10.5）（P < 0.05）。