海绵体神经损伤性勃起功能障碍治疗研究进展

随着前列腺手术的广泛开展和骨盆骨折尿道损伤患者的不断增多,海绵体神经损伤性阴茎勃起功能障碍越来越受到关注。海绵体神经损伤后,阴茎平滑肌和内皮细胞凋亡,NOS阳性神经密度降低,进而出现海绵体平滑肌纤维化。损伤后的神经再生策略一直是ED研究的热点之一。本综述将围绕促进海绵体神经再生的治疗策略,讨论海绵体神经损伤性ED治疗的基础及临床研究现状,既涉及神经营养因子、RhoA/ROCK抑制剂、亲免素配体、促红细胞生成素、干细胞治疗、基因治疗等传统策略,也包含富血小板血浆和低强度体外冲击波治疗等易于临床转化的新的治疗方式。本综述旨在为相关领域学者提供参考,以期开展更多具有较高临床转化意义的研究。     

干细胞治疗勃起功能障碍研究进展

勃起功能障碍(ED)的发生与血管内皮功能障碍及相关神经的损伤有关。近年来,干细胞对阴茎勃起神经和海绵体血管内皮细胞修复保护作用的临床前研究已成为研究的热点。早期研究显示干细胞或基因修饰的干细胞对ED治疗持久有效,并有可能成功治愈ED。间充质干细胞、肌源性干细胞、胚胎干细胞、脂肪源性干细胞、内皮祖细胞等均具有不同的分化潜能,在内皮细胞的更新、修复及神经组织细胞的保护方面有各自的优势。干细胞有望用于人类ED的治疗。     

骨髓间充质干细胞即刻与延时治疗修复阴茎海绵体神经损伤大鼠勃起功能的研究

目的:拟观察在双侧阴茎海绵体神经(CN)损伤的大鼠神经性勃起功能障碍(NED)模型中即刻和延时向阴茎海绵体注射骨髓间充质干细胞(BM-MSCs)对大鼠阴茎勃起功能的修复作用。方法:选取28只8周龄、体重200~250 g雄性SD大鼠随机分为4组:假手术组找到双侧海绵体神经后不做处理直接关腹并缝合皮肤;对照组、即刻治疗组和延时治疗组均通过钳夹的方式损伤双侧阴茎海绵体神经建立NED模型随后关腹缝合。假手术组和对照组向阴茎海绵体内注射对照剂,即刻治疗组注射BM-MSCs,延时治疗组术后两周注射BM-MSCs。术后12周采用电刺激CN记录阴茎海绵体内压(ICP),颈动脉穿刺测定平均动脉压(MAP),ICP/MAP作为勃起功能的评价指标来评估大鼠的勃起功能。在勃起功能检测后处死大鼠,取阴茎海绵体中段组织检测平滑肌、胶原和神经纤维。结果:在手术后12周,即刻治疗组和延时治疗组ICP和ICP/MAP均较对照组升高(P0.05)。即刻治疗组和延时治疗能提高大鼠海绵体组织内平滑肌与胶原的比例(P0.05),同时两组阴茎海绵体内平滑肌含量高于对照组(P0.05),阴茎海绵体背神经内神经微丝(NF)蛋白阳性神经纤维数目和nNOS的表达高于对照组(P0.05)。结论:阴茎海绵体内注射BM-MSCs能对双侧CN损伤大鼠的勃起功能恢复有促进作用。BM-MSCs治疗可以提高海绵体组织内平滑肌与胶原的比例,改善纤维化,并能提高海绵体组织中平滑肌含量、阴茎背神经的神经丝含量和nNOS的表达水平,术后延时治疗同样能取得一定的治疗效果。     

血管性勃起功能障碍的诊疗现状及进展

勃起功能障碍(erectile dysfunction,ED)是指持续或反复出现的阴茎不能达到和(或)维持勃起以完成满意的性交,且病程维持至少3个月[1]。阴茎勃起过程是神经发动、动脉供血与海绵体储血的综合结果,任何一个环节出现问题都将导致ED。血管性ED可由于动脉血流入的减少,海绵体平滑肌(smooth muscle cell SMC)舒张能力减弱,静脉闭合功能障碍和海绵体纤维化等病理变化影响到阴茎勃起。血管性ED在ED病人中十分多见,近年来勃起机制的探明使血管性ED诊疗有了突飞猛进的发展,本文就此作一综述。     

骨盆骨折并后尿道损伤致阴茎勃起功能障碍28例临床分析

目的探讨骨盆骨折并后尿道损伤致阴茎勃起功能障碍（ED）的类型、发病机制和治疗方法。方法通过病史、查体、实验室检查，国际勃起功能指数，NPT、罂粟碱试验、阴茎Doppler声、动态阴茎海绵体造影、球海绵体肌反射和尿动力学检查等方法，对骨盆骨折并后尿道损伤引起的阴茎勃起功能障碍28例患者进行诊断、分类，对不同类型的ED患者进行相应治疗。站果ED28例，其中神经血管性ED22例，血管性ED6例。22例神经血管性ED化学假体治疗，22例注药后阴茎勃起完成性交。其中5例治疗半年后阴茎勃起功能逐渐恢复；17例未能坚持治疗，12例改服万艾克，5例服药后完成性交。4例血管性ED行腹壁下动脉与阴茎背深静脉吻合术，术后能完成性交。结论ED是骨盆骨折并后尿道损伤的常见并发症。神经血管损伤是ED的主要原因，化学假体和口服万艾克有一定疗效。血管性ED手术治疗有一定疗效。     

阴茎勃起功能障碍的致病因素及治疗的现状

<正>阴茎勃起功能障碍(ED)是一种较常见的男性疾病,据调查约40%的男性长期或短期承受过ED的苦恼~([1])。阴茎勃起过程涉及神经、内分泌、血管系统、激素水平、心理状态等诸多因素。本文在简介阴茎勃起结构和勃起机制基础上综述了ED的病因及治疗方法的现状。一、阴茎的勃起结构及勃起机制(一)阴茎的勃起结构阴茎的勃起结构是一对阴茎海绵体及一条尿道海绵体。每条海绵体均由致密结缔组织膜(白膜)     

睾酮对大鼠阴茎海绵体平滑肌细胞增殖的影响及其与雄激素受体水平变化的关系

睾酮缺乏是阴茎勃起功能障碍(ED)可能的原因之一[1].为探讨睾酮在阴茎海绵体生长发育中的作用,我们设想阴茎海绵体平滑肌细胞(CCSM)中存在雄激素受体(AR),接受体内睾酮的刺激作用.     

海绵体神经损伤所致ED大鼠模型建立

目的 :寻找大鼠海绵体神经并建立神经损伤所致ED大鼠模型。　方法 :对 2 0只大鼠进行解剖 ,在外科显微镜下找到海绵体神经并经电刺激试验证实。随后将 4 2只实验大鼠随机分为假手术对照组、单侧海绵体神经损伤组及双侧海绵体神经损伤组。术后 3周用阿朴吗啡试验来评估所建动物模型。　结果 :盆大神经节位于背侧前列腺后外侧叶表面 ,海绵体神经是最大的传出神经。诱发阴茎勃起的电刺激参数 :电压 5V、刺激频率 2 0Hz及刺激时间 5ms。术后 3周 ,阿朴吗啡均能诱发对照组大鼠阴茎勃起 ,30min内平均勃起 (2 5 7± 1 4 0 )次。实验组大鼠 ,无论单侧损伤还是双侧损伤 ,均丧失勃起功能 (0 0 0± 0 0 0 )。　结论 :大鼠较大的盆大神经节及海绵体神经易于辨认 ,电刺激反应明显 ,是建立海绵体神经损伤性ED模型的理想动物。无论是单侧海绵体神经损伤还是双侧海绵体损伤 ,损伤早期 ,大鼠均丧失勃起功能     

勃起功能障碍患者及不同年龄与正常人阴茎海绵体平滑肌组织学观察

目的：了解正常及勃起功能障碍（ED）患者阴茎海绵体的差异和改变对阴茎勃起的影响。方法：取10例不同年龄正常及ED病人阴茎海绵体组织，镜下观察阴茎海绵体结构变化。结果：3例ED病人阴茎海绵体平滑肌细胞及弹力纤维减少。老年性阴茎海绵体平滑肌及弹力纤维明显减少。青壮年阴茎海绵体平滑肌细胞及弹力纤维极为丰富。结论：阴茎海绵体结构改变对阴茎勃起功能有较大的影响。     

高血压大鼠海绵体平滑肌细胞间连接的变化

目的:比较自发性高血压大鼠(SHR)和正常血压大鼠阴茎海绵体平滑肌细胞间连接改变及与阴茎勃起功能的关系。方法:注射阿朴吗啡(APO)观察14周龄SHR(SHR组,n=5)、W istar-Kyoto大鼠(WKY组,n=5)阴茎勃起情况,用透射电镜观察其阴茎海绵体平滑肌细胞间连接超微结构,RT-PCR测定海绵体平滑肌细胞Connexin 43的mRNA表达,免疫组化观察Connexin 43蛋白表达。结果:SHR组大鼠阴茎勃起次数明显低于WKY组(P<0.05),电镜发现SHR组大鼠阴茎海绵体平滑肌细胞间大量胶原纤维增生,Connexin 43蛋白及其mRNA表达较WKY组显著降低(P<0.05)。结论:高血压影响阴茎勃起功能,阴茎海绵体平滑肌细胞间连接的病理改变可能是高血压性勃起功能障碍的发病机制之一。     

他达拉非治疗ED的概述

勃起功能障碍(ED)是男科常见病、多发病,PDE5抑制剂能竞争性抑制PDE5而抑制cGMP的水解,提高阴茎海绵体平滑肌细胞内cGMP浓度,达到治疗ED的效果。他达拉非可使阴茎海绵体内cGMP水平提高,从而导致勃起。他达拉非可有效改善各种病因和各种程度ED患者的勃起功能;其具有以下特点:有效时间长,安全性高,易于被患者及性伴侣接受,增强性自信、性自尊,性体验更加自然。使患者从生理、心理上最大程度的得到治疗,有效提高性生活质量。因此他达拉非在ED患者的治疗中值得推广。     

Potential differentiation of human mesenchymal stem cell transplanted in rat corpus cavernosum toward endothelial or smooth muscle cells

One of the causes of erectile dysfunction (ED) is the damaged penile cavernous smooth muscle cells (SMCs) and sinus endothelial cells (ECs). To investigate the feasibility of applying immortalized human mesenchymal stem cells (MSCs) to penile cavernous ECs or SMCs repair in the treatment of ED, the in vivo potential differentiation of the immortalized human MSCs toward penile cavernous endothelial or smooth muscle was investigated. One clone of immortalized human bone marrow mesenchymal stem cell line B10 cells via retroviral vector encoding v-myc were transplanted into the cavernosum of the Sprague-Dawley rats and harvested 2 weeks later. The expression of CD31, von Willebrand factor (vWF), smooth muscle cell actin (SMA), calponin and desmin was determined immunohistochemically in rat penile cavernosum. Multipotency of B10 to adipogenic, osteogenic or chondrogenic differentiation was found. Expression of EC specific markers (CD31 or vWF protein) and expression of SMC specific markers (calponin, SMA or desmin protein) were demonstrated in grafted B10 cells. When human MSCs were transplanted into the penile cavernosum, they have the potential to differentiate toward ECs or SMCs. Human MSCs may be a good candidate in the treatment of penile cavernosum injury.     

Human neural crest stem cells transplanted in rat penile corpus cavernosum to repair erectile dysfunction

OBJECTIVE

To investigate the feasibility of applying neural crest stem cells (NCSCs), with multipotent capacity, to repair injury in the penile cavernosum, the HNC10.K10 (K10) immortalized NCSC line was transplanted into the penile cavernosum of adult rats, as one of the causes of erectile dysfunction is damaged penile cavernous smooth muscle cells and sinus endothelial cells.

MATERIALS AND METHODS

The K10 human NCSC line was generated via transfection of primary cultured NCSC with a retroviral vector encoding v‐myc. K10 NCSCs were transplanted into the cavernosum of adult rats. The expression of cell type‐specific markers for endothelial cells (CD31 and von Willebrand factor), and specific markers for smooth muscle cells (smooth muscle cell actin, calponin, and desmin) was determined immunohistochemically in the penile cavernosum of rats 2 weeks after transplantation.

RESULTS

In the rat cavernosum, transplanted K10 NCSCs identified by human nuclear antigen labelling expressed cell type‐specific markers for endothelial cells (CD31 and von Willebrand factor), and specific markers for smooth muscle cells (smooth muscle cell actin, calponin, and desmin) 2 weeks after transplantation. Human NCSCs transplanted into the rat penile corpus cavernosum differentiated into endothelial cells or smooth muscle cells, as shown by their expression of cell type‐specific markers for the cell types.

CONCLUSION

It appears that NCSCs are an ideal cell source for reconstructing endothelial and smooth muscle cells in the corpus cavernosum in cell therapy for patients with erectile dysfunction.     

短发夹RNA对大鼠阴茎海绵体平滑肌细胞磷酸二酯酶5型基因表达的抑制作用

目的 观察短发夹RNA(shRNA)对大鼠阴茎海绵体平滑肌细胞磷酸二酯酶5型(PDE5)基因表达的抑制作用,探讨运用RNA干扰(RNAi)技术治疗勃起功能障碍(ED)的可行性.方法 构建靶向大鼠PDE5基因的shRNA重组腺病毒rAd-rPDE5-shRNA,将其转染大鼠阴茎海绵体平滑肌细胞48 h后,通过荧光标签进行显微计数确定转染效率,并以逆转录-聚合酶链反应(RT-PCR)及Western blot检测PDE5基因的表达水平.结果 rAd-rPDE5-shRNA构建成功,转染大鼠阴茎海绵体平滑肌细胞效率达95%以上,并使PDE5基因表达在mRNA水平抑制(80.78±2.30)%,在蛋白水平抑制(67.39±3.33)%.结论 以腺病毒为载体表达的shRNA能稳定、有效地抑制大鼠阴茎海绵体平滑肌细胞PDE5基因的表达.     

Expression of functional prostaglandin D (DP) receptors in human corpus cavernosum smooth muscle

Prostaglandin D(2) (PGD(2)) binds to specific G-protein coupled receptors (DP) and induces smooth muscle relaxation by stimulating the synthesis of intracellular cAMP. In this study, we examined the role of PGD(2) and DP receptors in regulating human penile smooth muscle contractility. We determined that human corpus cavernosum tissue and smooth muscle cells in culture expressed functional DP receptor and lipocalin-like prostaglandin D synthase by reverse-transcribed polymerase chain reaction (RT-PCR). Functional PGD synthase activity was confirmed by the synthesis of PGD(2) in human corpus cavernosum smooth muscle cells upon addition of exogenous arachidonic acid. Organ bath preparations of human corpus cavernosum tissue strips, contracted with phenylephrine, relaxed in a dose-dependent fashion to either PGD(2) or the DP selective agonist BW245C. Cultures of human corpus cavernosum smooth muscle cells treated with BW245C showed a two-fold increase in cAMP synthesis. These data are consistent with the expression of functional DP receptors in human corpus cavernosum. This suggests the presence of an intact prostanoid autocrine system that may play a role in regulating penile erectile function.     

Effects of integrin‐linked kinase on human corpus cavernosum smooth muscle cell cytoskeletal organisation

We investigated the effects of integrin‐linked kinase (ILK) on the in vitro attachment, spreading, migration and microfilament dynamics of human corpus cavernosum smooth muscle cells. ILK small interfering RNA (siRNA) was used to transfect human corpus cavernosum smooth muscle cells; and cell attachment, spreading and migration were assessed. Additionally, microfilament dynamics were evaluated using Alexa Fluor 488 and phalloidin staining. We found that ILK gene knock‐down significantly inhibited human corpus cavernosum smooth muscle cell attachment, spreading and migration. Moreover, blocking the expression of ILK disturbed actin cytoskeleton reorganisation and morphology in human corpus cavernosum smooth muscle cells. These results show that the targeting of ILK with siRNA significantly inhibited cell attachment, spreading, migration and microfilament dynamics in human corpus cavernosum smooth muscle cells. These findings indicate that ILK might be a potential therapeutic molecular target for the treatment of erectile dysfunction.     

Fibrosis of corpus cavernosum in animals following cavernous nerve ablation

Aim: To investigate alterations of smooth muscle celis and collagen fibers in corpus cavernosum following cavernous neurectomy and its relation to the expression of transforming growth factor-β1 (TGF-β1). Methods: Ten adult male SD rats (neurectomy group) were subject to a bilateral cavernous nerve (CN) resection aseptically under an operating microscope, with 6 sham-operated rats as the control. Fifteen weeks after the operation, the penile speci mens were collected and prepared for quantitative-analyzing of ratio of smooth muscle to collagen fibers in corpus cavernosum with confocal microscopy, and for detecting the expression of TGF-β1 by RT-PCR and western-blot. Resulte: Smooth muscle celis that show red color after fluorescent-labeling with tetramethylrhodamine isothiocyanate phalloidin and collagen fibers that produce green autofluorescence after paraformaldehyde fixation were clearly iden tified     

Activation of soluble guanylate cyclase causes relaxation of corpus cavernosum tissue: synergism of nitric oxide and YC-1

Nitric oxide (NO) activates corpus cavernosum smooth muscle soluble guanylate cyclase (sGC) and increases the synthesis of cGMP that results in smooth muscle relaxation and ultimately, penile erection. To characterize sGC and define the potential synergy between NO and the allosteric activator YC-1 in corpus cavernosum, rat sGC was activated by either sodium nitroprusside (SNP) or YC-1, and YC-1 potentiated the effects of SNP with a 200-fold activation of sGC. Both SNP and YC-1 decreased the Km and increased the Vmax. ODQ significantly inhibited sGC activated by SNP with IC50 of 0.5 nM, but did not affect the sGC activated by YC-1 as well as basal sGC activity. SNP and YC-1 synergistically increased intracellular cGMP levels in rabbit corpus cavernosum smooth muscle cell cultures. YC-1 significantly relaxed rabbit cavernosum tissue strips in organ baths with an EC50 of 8.4 microM. In the presence of L-nitroarginine methyl ester to block endogenous NO production, co-administration of SNP shifted the dose response of YC-1 to the left, showing the synergism of SNP and YC-1 in tissue strips. In view of the clinical efficacy of phosphodiesterase-5 inhibitors, activation of sGC may provide an alternative means for enhancing the activity of neurally derived NO during sexual stimulation in the corpus cavernosum, representing a novel approach for the treatment of erectile dysfunction.     

PDE5基因siRNA对人阴茎海绵体平滑肌细胞cGMP的影响

目的:观察PDE5基因小干扰RNA(siRNA)对人阴茎海绵体平滑肌细胞环磷酸鸟苷(cGMP)的影响,为阴茎勃起功能障碍(ED)的基因治疗提供实验依据。方法:使用美国Amb ion公司提供的设计软件设计并合成人PDE5基因的siRNA序列,合成3对PDE5 siRNA和1对阴性对照siRNA,转染人阴茎海绵体平滑肌细胞,同时设定空白转染组为对照组。以酶联免疫法分别检测转染后不同时间(24、48、72、96 h)点海绵体平滑肌细胞内cGMP浓度变化,观察PDE5 siRNA对海绵体平滑肌细胞内cGMP的影响。结果:siRNA1、siRNA2和siRNA3转染后人阴茎海绵体平滑肌细胞cGMP水平显著高于阴性对照siRNA组和空白对照组(P<0.05),在转染后72 h最为显著,siRNA1、siRNA2、siRNA3、阴性对照siRNA和空白对照组cGMP测定值分别为:5.89±0.19、3.52±0.16、2.88±0.08、0.72±0.12、0.60±0.16 pmol/m l,siRNA1组明显高于siRNA2、siRNA3组(P<0.05)。结论:体外化学合成的PDE5 siRNA能有效地增加海绵体平滑肌细胞内cGMP的水平,不同序列siRNA增加cGMP水平的能力不同,为ED的基因治疗提供了新思路。