Cho Min Chul, Park Kwanjin, Kim Soo Woong, Paick Jae-Seung
Department of Urology, Dongguk University College of Medicine, Goyang, Republic of Korea.
Department of Urology, Seoul National University College of Medicine, Seoul, Republic of Korea.
J Urol. 2015 May;193(5):1716-23. doi: 10.1016/j.juro.2014.10.099. Epub 2014 Oct 27.
We determined whether Rho-kinase inhibition would improve corporal veno-occlusive dysfunction by suppressing apoptosis and fibrosis via normalization of the Rho-kinase driven pathways related to the 2 structural alterations in a rat model of cavernous nerve crush injury.
A total of 30 male 10-week-old male Sprague Dawley® rats were equally divided into 3 groups, including sham surgery, cavernous nerve crush injury and cavernous nerve crush injury treated with fasudil. The treated group received fasudil (30 mg/kg) daily for 4 weeks starting day 1 postoperatively. Electrostimulation and dynamic infusion cavernosometry were performed 4 weeks postoperatively. Penile tissue was processed for imm unohistochemistry, double immunofluorescent and Masson trichrome staining, TUNEL, caspase-3 activity assay and Western blot.
The cavernous nerve crush injury group showed significantly lower intracavernous pressure/mean arterial pressure, and higher maintenance and drop rates than the sham surgery group. Rho-kinase inhibition in the injury plus fasudil group restored erectile responses and dynamic infusion cavernosometry parameters. Increased apoptosis, decreased immunohistochemical staining of α-SMA and increased caspase-3 activity were noted in the injury group. In that group densitometry revealed increased ROCK1 expression, increased MYPT1 phosphorylation, decreased Akt phosphorylation, decreased Bad phosphorylation and a decreased Bcl2-to-Bax ratio. A significantly decreased smooth muscle-to-collagen ratio and increased fibroblast pCofilin were also observed in the injury group, as was increased phosphorylation of cofilin, a downstream effector of LIMK2. Rho-kinase inhibition in the injury plus fasudil group alleviated the histological and molecular dysregulation.
Our data suggest that early inhibition of Rho-kinase after cavernous nerve crush injury may prevent corporal apoptosis and fibrosis by suppressing the Akt/Bad/Bax/caspase-3 and LIMK2/cofilin pathways, preventing corporal veno-occlusive dysfunction and erectile dysfunction.
我们通过在海绵体神经挤压损伤大鼠模型中，使与两种结构改变相关的Rho激酶驱动通路正常化，来确定抑制Rho激酶是否会通过抑制细胞凋亡和纤维化来改善海绵体静脉闭塞功能障碍。
总共30只10周龄雄性Sprague Dawley®大鼠被平均分为3组，包括假手术组、海绵体神经挤压损伤组和用法舒地尔治疗的海绵体神经挤压损伤组。治疗组从术后第1天开始每天接受法舒地尔（30 mg/kg）治疗，持续4周。术后4周进行电刺激和动态灌注海绵体测压。阴茎组织进行免疫组织化学、双重免疫荧光和Masson三色染色、TUNEL、半胱天冬酶-3活性测定和蛋白质印迹分析。
与假手术组相比，海绵体神经挤压损伤组的海绵体内压/平均动脉压显著降低，维持率和下降率更高。损伤加用法舒地尔组的Rho激酶抑制恢复了勃起反应和动态灌注海绵体测压参数。损伤组细胞凋亡增加，α-SMA免疫组织化学染色减少，半胱天冬酶-3活性增加。在该组中，密度测定显示ROCK1表达增加、MYPT1磷酸化增加、Akt磷酸化减少、Bad磷酸化减少以及Bcl2与Bax的比率降低。损伤组还观察到平滑肌与胶原蛋白的比率显著降低，成纤维细胞pCofilin增加，以及LIMK2的下游效应物cofilin的磷酸化增加。损伤加用法舒地尔组的Rho激酶抑制减轻了组织学和分子失调。
我们的数据表明，海绵体神经挤压损伤后早期抑制Rho激酶可能通过抑制Akt/Bad/Bax/半胱天冬酶-3和LIMK2/cofilin通路来预防海绵体细胞凋亡和纤维化，从而预防海绵体静脉闭塞功能障碍和勃起功能障碍。
