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Published on: September 8, 2016
Neuronal Nitric Oxide Synthase Gene Transfer into the Rat Prostate Using In Vivo Electroporation
Masayuki Otani1, Masaki Yoshida1, Koichi Masunaga1
1Department of Urology, Nishi Kumamoto Hospital, Kumamoto, JapanDepartment of Urology, Kumamoto Rosai Hospital of Japan Labor Health and Welfare Organization, Kumamoto, JapanDepartment of Urology, Tokyo Metropolitan Geriatric Hospital, Tokyo, JapanDepartment of Urology, Toshiba Hospital, Tokyo, JapanDepartment of Urology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, JapanDepartment of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Objectives:
It has been reported that nitric oxide (NO) mainly contributes to prostate or urethral smooth muscles relaxation, and that nitrergic innervation and neuronal NO synthase (nNOS) levels are decreased in benign prostatic hyperplasia. The purpose of the present study was to evaluate the feasibility to gene therapy for benign prostatic hyperplasia by transferring nNOS gene into the rat prostate with in vivo electroporation (EP) procedure.
Methods:
Male Sprague-Dawley rats were divided into four groups (sham, only EP, only nNOS injection, and nNOS gene injection with EP groups). Fifty micrograms of luciferase gene and nNOS expression vectors in 50 µL of K-PBS (potassium-phosphate buffered saline) were injected into the prostate. Immediately after the injection of these vectors, the vector injection points were electroporated by the two-square parallel electrodes. Two days after gene transfer, luciferase analysis and an immunohistochemical staining for nNOS were performed, and NO2 (-) /NO3 (-) (NOX ) release was measured using high-performance liquid chromatography coupled with the microdialysis procedure.
Results:
The optimal electric pulse conditions were 50 V, 1 Hz and 10 msec. In vivo EP with these conditions showed the increase in the luciferase gene expression approximately 300-fold of the control group. In the nNOS gene injection with EP group, the marked nNOS immunoreactivity was observed, and NOX release was significantly higher, as compared to other groups.
Conclusion:
The results suggest that EP is a feasible technique for in vivo gene transfer into the rat prostate, and that the transferred nNOS gene functionally expresses and contributes to NO production.
