COX-2-10aa-PGIS 基因治疗可改善海绵体神经损伤大鼠的勃起功能。

COX-2-10aa-PGIS gene therapy improves erectile function in rats after cavernous nerve injury.

机构信息

Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, Nanjing, China.

出版信息

J Sex Med. 2013 Jun;10(6):1476-87. doi: 10.1111/jsm.12147. Epub 2013 Apr 3.

DOI:10.1111/jsm.12147

PMID:23551902

Abstract

INTRODUCTION

Erectile dysfunction (ED) is a very common complication after radical prostatectomy. COX-2-10aa-PGIS is a newly engineered protein with COX-2 and prostacyclin synthase activities that converts arachidonic acid directly to prostacyclin (prostaglandin I2 [PGI2]). PGI2 is a potent smooth muscle relaxant.

AIM

The purpose of this study was to explore the effect and mechanism of COX-2-10aa-PGIS gene therapy in penile rehabilitation.

METHODS

Bilateral cavernous nerve crush (BCNC) in adult Sprague-Dawley rats was used to mimic radical prostatectomy-induced ED. Sprague-Dawley rats were randomly assigned into four groups: 1. sham surgery; 2. BCNC; 3. BCNC + null control recombinant adenovirus intracavernous injection; and 4. BCNC + Ad-COX2-10aa-PGIS intracavernous injection. Twenty-eight days later, intracavernosal pressure (ICP) was recorded under cavernous nerve stimulation; in the meantime, the mean arterial pressure (MAP) was monitored. At the end of the measurement, the penis was harvested and processed for (i) immunohistochemistry analysis of endothelial nitric oxide synthase (eNOS), alpha-smooth muscle actin (α-SMA), and transforming growth factor beta-1 (TGF-β1); (ii) Masson's trichrome stain for smooth muscle/collagen ratios; (iii) Western blot of eNOS, α-SMA, TGF-β1, and COX2-10aa-PGIS; and (iv) terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay for apoptosis.

MAIN OUTCOME MEASURES

Erectile function was evaluated by ICP/MAP. Smooth muscle and endothelium functions in corpora cavernosum were assessed by Masson's trichrome stain, immunohistochemistry, and Western blot. Apoptosis was identified by TUNEL assay.

RESULTS

The results were the following: 1. COX2-10aa-PGIS gene therapy improved erectile function (82%, compared with control) in the BCNC rat model; 2. COX2-10aa-PGIS gene therapy increased eNOS (121%) and α-SMA (118%) expression and decreased TGF-β1 (45%) expression; 3. COX2-10aa-PGIS gene therapy reduced cell apoptosis after cavernous nerve injury (64%); and 4. COX2-10aa-PGIS gene therapy improved smooth muscle/collagen ratios (81%).

CONCLUSION

Our data demonstrated that COX2-10aa-PGIS improved erectile function after cavernous nerve injury through antifibrotic and anti-apoptotic mechanisms.

摘要

简介

勃起功能障碍（ED）是根治性前列腺切除术后非常常见的并发症。COX-2-10aa-PGIS 是一种新工程化的蛋白质，具有 COX-2 和前列环素合酶活性，可将花生四烯酸直接转化为前列环素（前列腺素 I2 [PGI2]）。PGI2 是一种有效的平滑肌松弛剂。

目的

本研究旨在探讨 COX-2-10aa-PGIS 基因治疗在阴茎康复中的作用和机制。

方法

采用成年 Sprague-Dawley 大鼠双侧海绵体神经挤压（BCNC）模拟根治性前列腺切除术后 ED。Sprague-Dawley 大鼠随机分为四组：1.假手术；2. BCNC；3. BCNC+无效对照重组腺病毒海绵体内注射；和 4. BCNC+Ad-COX2-10aa-PGIS 海绵体内注射。28 天后，在海绵体神经刺激下记录海绵体内压（ICP）；同时监测平均动脉压（MAP）。测量结束时，采集阴茎并进行以下处理：（i）内皮型一氧化氮合酶（eNOS）、α-平滑肌肌动蛋白（α-SMA）和转化生长因子β1（TGF-β1）的免疫组织化学分析；（ii）平滑肌/胶原比的 Masson 三色染色；（iii）eNOS、α-SMA、TGF-β1 和 COX2-10aa-PGIS 的 Western blot；和（iv）末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）测定凋亡。

主要观察指标

ICP/MAP 评估勃起功能。通过 Masson 三色染色、免疫组织化学和 Western blot 评估海绵体平滑肌和内皮功能。通过 TUNEL 测定鉴定细胞凋亡。

结果

结果如下：1. COX-2-10aa-PGIS 基因治疗改善了 BCNC 大鼠模型的勃起功能（82%，与对照组相比）；2. COX-2-10aa-PGIS 基因治疗增加了 eNOS（121%）和 α-SMA（118%）的表达，降低了 TGF-β1（45%）的表达；3. COX-2-10aa-PGIS 基因治疗减少了海绵体神经损伤后的细胞凋亡（64%）；和 4. COX-2-10aa-PGIS 基因治疗改善了平滑肌/胶原比（81%）。