COX-2-10aa-PGIS Gene Therapy Improves Erectile Function in Rats after Cavernous Nerve Injury
COX-2-10aa-PGIS Gene Therapy Improves Erectile Function in Rats after Cavernous Nerve Injury
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COX-2-10aa-PGIS 基因疗法改善海绵神经损伤后大鼠的勃起功能
DOI：
10.1111/jsm.12147
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发表时间：
2013-06-01
影响因子：
3.5
通讯作者：
Wang, Run
中科院分区：
文献类型：
--
作者：
Lin, Haocheng;Yuan, Jiuhong;Wang, Run
导言。勃起功能障碍(ED)是前列腺癌根治术后常见的并发症。COX-2-10aa-PGIS是一种具有COX-2和前列环素合成酶活性的新型工程蛋白，可将花生四烯酸直接转化为前列环素(PGI2)。前列腺素I2是一种有效的肌松药。瞄准。本研究旨在探讨COX-2-10aa-PGIS基因治疗在阴茎康复中的作用及其机制。方法：研究方法。采用成年SD大鼠双侧海绵体神经挤压模型，模拟前列腺癌根治术诱发的ED。将SD大鼠随机分为4组：假手术组、BCnC组、BCnC+空对照重组腺病毒海绵体注射组和BCnC+Ad-COX2-10aa-PGIs海绵体注射组。28d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)。测量结束后，取阴茎，进行(I)内皮型一氧化氮合酶(ENOS)、α-平滑肌肌动蛋白(-SMA)和转化生长因子-β1(TGF-1)的免疫组织化学分析；(Ii)Masson氏三色染色检测平滑肌/胶原蛋白比值；(Iii)Western印迹法检测eNOS、-SMA、TGF-1和COX2-10aa-PGIs；(Iv)末端脱氧核苷酸转移酶dUTP缺口末端标记法(TUNEL)检测细胞凋亡。主要观察指标。勃起功能评定采用ICPMAP。用Masson‘s三色染色、免疫组织化学和Western印迹法检测海绵体的平滑肌和内皮功能。TUNEL法检测细胞凋亡。结果。研究结果如下：1.COX2-10aa-PGIs基因治疗可明显改善BCNC模型大鼠的勃起功能(与对照组相比提高82%)；2.COX2-10aa-PGIs基因治疗可增加eNOS(121%)和-SMA(118%)的表达，降低转化生长因子-1(45%)的表达；3.COX2-10aa-PGIs基因治疗可减少海绵体神经损伤后的细胞凋亡(%)；结论。我们的数据表明，COX2-10aa-PGIs通过抗纤维化和抗细胞凋亡机制改善海绵体神经损伤后的勃起功能。
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 (-SMA), and transforming growth factor beta-1 (TGF-1); (ii) Masson's trichrome stain for smooth muscle/collagen ratios; (iii) Western blot of eNOS, -SMA, TGF-1, 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 -SMA (118%) expression and decreased TGF-1 (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.