阴茎海绵体平滑肌细胞表型转化及其影响因子的研究进展

阴茎海绵体平滑肌细胞是组成阴茎海绵体的主要功能成分,其表型转化是平滑肌细胞增殖和迁移的关键性起始步骤。因此,探讨平滑肌细胞表型转化的机制及其影响因子在阴茎勃起功能障碍的防治过程中具有重要意义。目前通常将平滑肌细胞分为收缩型(分化型)和合成型(未分化型、增殖型或去分化型)两种类型,并发现L转化生长因子(TGF-β)、转录因子E2F1、基本转录元件结合蛋白2(BTEB2)、胰岛素等因素可能影响平滑肌细胞表型转化。本文就近年来阴茎海绵体平滑肌细胞表型转化及其影响因子的研究进展作一简要综述。     

阴茎海绵体神经重建术研究进展

阴茎海绵体神经损伤是导致勃起功能障碍(ED)的常见原因,海绵体神经重建修复有望恢复患者性功能。海绵体神经重建的方法有直接吻合、自体神经移植以及可降解生物材料的替代,目前应用于临床的只有自体腓肠神经移植。神经重建过程中,神经生长因子也起重要作用。本文就近年阴茎海绵体神经的重建方法作一综述。     

海绵体脚结扎术后阴茎持续勃起

双侧阴茎海绵体脚结扎辅以阴茎背深静脉结扎,是治疗静脉性勃起功能障碍(ED),较有效的方法之一。目前研究静脉性ED的手术方法和疗效较多,对术后并发症及其处理的研究则较少。本文对本科1995年1月至2001年6月收治的31例静脉性ED患者,在行海绵体脚结扎术后发生阴茎持续勃起状况及处理方法进行分析、讨论。现报告如下。     

B超下阴茎海绵体"繁星征"与 ED

目的 探讨B超下阴茎海绵体出现的“繁星征”与ED的关系,研究其发病机制。方法 30例ED患者行阴茎海绵体B超检查,并与30例无ED志愿者行B超对照。另对l例ED“繁星征”患者行阴茎海绵体病理检查。结果 30例ED患者中,29例发现有B超下阴茎海绵体“繁星征”。其中26例全阴茎弥散分布强回声光点,为临床诊断的中、重度ED患者（IIEF-5评分〈11分）。另3例仅局限在一部分,且回声光点较少,为轻度ED（IIEF-5评分12～19分）。而无ED志愿者无此现象。1例“繁星征”患者病理检查为阴茎海绵体胶原纤维增生伴玻璃样变。结论 阴茎海绵体广泛纤维化是出现B超下“繁星征”的病理基础。因限制海绵窦充盈而影响阴茎勃起。外伤与炎症是其致病因素。     

成人及家兔阴茎海绵体平滑肌细胞培养和鉴定

目的 :探索阴茎海绵体平滑肌细胞 (CCSM)体外培养及鉴定方法。　方法 :采用勃起功能正常的成年男子及新西兰家兔的新鲜阴茎标本 ,经处理后采用原代细胞培养法 ,对阴茎海绵体组织块进行培养 ,获得CCSM后用光镜、透射电镜和免疫组化等多种方法从细胞形态学及CCSM生长特性等不同侧面对培养的人及新西兰家兔的CCSM进行鉴定 ,并测定CCSM在体外培养时的细胞增殖情况。　结果 :经 2 4h潜伏期 ,体外培养的CCSM进入指数增长期 ,维持约 96h后进入平台期 ;从接种组织块至长成单层细胞约需 15～ 2 0d ,传代后约 4～ 7d后长成单层 ;传代后的细胞增殖旺盛 ,成束状生长 ,并呈现层叠排列及“峰 谷”现象 ;CCSM在体外可稳定传 7～ 12代 ,传代期间其形态及增殖活性变化不明显 ;各种检测鉴定均证实所获细胞为CCSM。　结论 :CCSM原代细胞培养法简便、稳定、可靠 ,对研究阴茎的勃起机制及勃起功能障碍的发生、发展、调控及其治疗药物筛选有重要理论及现实意义。     

降钙素基因相关肽对糖尿病性ED大鼠阴茎海绵体平滑肌细胞表型转化的影响

目的:了解降钙素基因相关肽(CGRP)对糖尿病性ED大鼠阴茎海绵体平滑肌细胞表型转化的影响。方法:利用链脲佐菌素建立糖尿病及糖尿病性ED大鼠模型。阴茎海绵体平滑肌细胞原代培养,并进行免疫细胞化学染色鉴定。实验分为2组:正常对照组和糖尿病性ED大鼠组。不同浓度(0、10,60,100 nmol/L)CGRP作用24h后,利用qRT-PCR检测各组细胞碱性调宁蛋白和骨桥蛋白mRNA的表达。结果:各组原代培养阴茎海绵体平滑肌细胞α-肌动蛋白阳性细胞率为(95.94±0.03)%。与正常对照组比较,糖尿病组ED大鼠阴茎海绵体平滑肌细胞碱性调宁蛋白mRNA表达显著减少(4.41±0.29 vs 10.35±0.62,P<0.01),而骨桥蛋白mRNA表达水平显著上调(5.28±0.32 vs 1.32±0.24,P<0.01)。当CGRP作用的终浓度为100 nmol/L时,糖尿病组大鼠阴茎海绵体平滑肌细胞经CGRP作用后,与未经其作用相比,碱性调宁蛋白mRNA表达显著上调(6.90±0.22 vs 4.41±0.29,P<0.01),而骨桥蛋白mRNA表达水平显著减少(3.26±0.31 vs 5.28±0.32,P<0.01)。结论:CGRP可使糖尿病性ED大鼠阴茎海绵体平滑肌细胞表型从合成型向收缩型转化。     

显微修复损伤的阴茎海绵体神经恢复大鼠勃起功能

目的　探讨利用显微外科技术修复外科损伤的阴茎海绵体神经 ,重建大鼠勃起通道的可行性。方法　 36只SD雄性大鼠随机平均分成 3组 ,即假手术对照组、双侧阴茎海绵体神经损伤组和显微修复组 ,术后 1、3个月后用阿朴吗啡 (APO)试验来评估所建动物模型 ,随后取阴茎干组织 ,利用NADPH d组化法证实nNOS阳性神经纤维的再生情况。结果　术后 1个月 ,神经部分切断组和显微修复组间差异无显著性 (P >0 .0 5 ) ;术后 3个月 ,APO所诱导的阴茎勃起试验中 ,显微修复组勃起率为 83 .3 % ;神经部分切断组勃起率一直为 0 % (P <0 .0 5 )。此外 ,神经套管组的nNOS阳性神经纤维数目在术后 3个月有显著增多 ,而神经部分切断组的nNOS阳性神经纤维数目同术后 1个月相比无增多 ,两两比较有差异有显著性 (P <0 .0 0 8)。结论　双侧阴茎海绵体损伤后 ,立即用显微外科技术吻合神经 ,是重建勃起通路 ,恢复勃起功能的一种有效方法。     

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。     

阴茎海绵体平滑肌钙信号通路研究进展

早在100多年前，研究者已经注意到Ca^2＋在生命中的作用，但直到1967年，美籍华人张槐耀发现钙调素后，人们才开始对Ca^2＋的作用机理有了深刻的认识，提出了Ca^2＋是胞内的一种第二信使。随着研究的深入，人们发现钙信号通路在生命活动中无处不在，具有非常重要的地位。阴茎勃起与疲软受很多因素的影响，但最终分子机制的共同通路就是钙信号通路。所以研究钙信号通路对阴茎勃起及勃起障碍的生理、病理、诊断及治疗等有很重要的意义。     

无创性动态阴茎海绵体测压初探

目的 :初步评价VISER软件进行无创性动态阴茎海绵体测压在ED诊断中的应用。　方法 :采用AquariusXLT型尿流动力学仪配置的VISER软件 ,辅助眼镜式影像仪听觉视觉性刺激 (AVSS)或阴茎海绵体内血管活性药物注射 (ICI)进行无创性动态海绵体测压 68例 ,其中ED病人 4 8例 ,正常对照 2 0例。　结果 :正常对照 2 0例均可通过AVSS诱发勃起 ;4 8例ED病人中 ,18例单纯通过AVSS可诱发勃起 ,占 3 7.5 % ;3 0例单纯AVSS无效者 ,给予罂粟碱 10mg海绵体内注射 ,其中 19例出现勃起 ,占 63 .3 %。VISER检查结果表明 ,除受ICI影响 ,ED罂粟碱组的勃起平均时间延长外 ,勃起数据和峰值数据中的其他多项指标ED各组均低于对照组 ,尤其表现为勃起和峰值总能量降低。　结论 :VISER具有精确的动态阴茎海绵体测压功能 ;辅助眼镜式影像仪有助于增强AVSS的效果和减少阴茎海绵体ICI剂量 ;由于VISER检查具有无创、准确、便捷等优点 ,有望成为今后ED诊断的首选方法。     

Apoptosis of corpus cavernosum in patients with organic erectile dysfunction

Objectives  It has been reported that apoptosis of penile erectile tissue occurs after penile denervation, castration, and diabetes mellitus in animal studies. Aim of this study was to investigate apoptosis in corpora cavernosa of patients with organic erectile dysfunction (ED). Methods  Cavernous biopsies were obtained from 38 patients with erectile dysfunction and 10 patients with normal erectile function. Apoptosis of tissues were determined via terminal deoxyuridine nucleotide end labeling method by using flow cytometry. Results  The mean ages of patients with ED and control patients were 50.65 ± 2.27, and 32.43 ± 2.90 years, respectively (P = 0.0001). Patients with ED were set in two groups as more than 50 years old and less than 50 years old for further analysis of age factor on apoptosis. The mean % apoptosis of ED patients was 26.22 ± 2.79 and control group was 11.26 ± 3.79, (P = 0.032). Mean fluorescence intensity (MFI) values were also 17.41 ± 3.21 and 6.59 ± 2.28, respectively (P = 0.039). MFI and % apoptosis values were not statistically significant different neither between the patients groups nor between the control and patients ≤50 years old (P > 0.05). Conclusions  We did not find any statistically significant difference with respect to apoptosis rates when we compared neither control group with ≤50 years old patients nor patients groups of ED. Because of this we did not have enough data to say that apoptosis has a prominent role on the development of ED independently from other factors. However, further studies are necessary to clarify the role of apoptosis in erectile dysfunction.     

Overactive corpus cavernosum: a novel cause of erectile dysfunction

Our recording of the electromyographic (EMG) activity of the corpus cavernosum (CC) in 59 patients with erectile dysfunction (ED) revealed 18 patients who had elevated electric activity, which presumably points to heightened tone of the CC smooth muscles. We investigated the hypothesis that this elevated EMG activity and muscular tone of the CC could be the cause of ED. The study comprised the said 18 subjects with the hypertonic CC muscles as study group (42.6 +/- 5.3 SD years), 15 healthy volunteers (41.8 +/- 5.1 SD years) and 15 patients (41.6 +/- 5.5 SD years) with ED who had not recorded elevated tone of the CC muscles as control group. The EMG activity was registered in the flaccid, erectile and detumescent phases by two electrodes inserted into the CC. Electrocavernosography (ECG) of healthy volunteers recorded in the flaccid phase showed regular slow waves (SW) and random action potentials (APs). The wave variables declined significantly in the erection phase (P < 0.01). In the study group, the SW variables in the flaccid phase increased significantly (P < 0.05) compared with the healthy volunteers and the rhythm was irregular. Erection did not occur with sildenafil but with intracavernosal injection of papaverine, which led to decline of the SW variables (P < 0.05). The control ED group exhibited in the flaccid phase diminished SW variables (P < 0.05) compared with the healthy volunteers. On erection with sildenafil administration, the SW variables showed significant reduction (P < 0.05). CC hypertonicity or 'overactive CC' was identified as a possible cause of ED. An elevated EMG activity of the CC muscle fibres in the flaccid phase presumably denotes hypertonicity of these fibres and their failure to relax to effect erection. The cause of elevated CCEMG activity and presumed muscle hypertonicity is unknown and could be functional or organic. Erection was produced by intracavernosus injection of papaverine and not by sildenafil. This condition of 'overactive CC' should be considered in the diagnosis of ED. However, further studies in the pathogenesis of the condition are warranted.     

Long-term results of corpus cavernosum autoinjection therapy for chronic erectile dysfunction

Between May 1985 and March 1992, 172 patients suffering from chronic erectile dysfunction (21-70 years old) underwent constant corpus cavernosum autoinjection therapy (CAT) with a standardized papaverine phentolamine mixture (16385 injections). Thereafter 41 patients continued CAT with the single agent papaverine (1257 injections). On the basis of both these 17642 protocol auto-injections, and over 6 years of experience with intracavernosal autoinjection therapy we conclude that, especially with the papaverine-phentolamine mixture, CAT constitutes an effective therapy (full rigidity in 95.8%) with tolerable side-effects for chronic erectile dysfunction when preceded by careful patient selection and thorough multi-disciplinary evaluation. This is especially so in the case of arterial and/or neurogenic aetiology of the erectile dysfunction. In addition, the contraindications must be strictly observed, the treatment and technique fully explained, and a regular follow-up instituted. CAT is generally well accepted by the patients and their partners (98.8%/97.6%) and has distinct positive effects on self-esteem (77.8%), performance anxiety (84.4%), and partnership (79.5%). The most serious side-effect was prolonged erection (25 out of 17642 injections). In 6 patients reversible fibrotic changes near the tunica albuginea were observed.     

Results and acceptance of corpus cavernosum autoinjection therapy in organic erectile dysfunction

Ambulatory autoinjection therapy of the corpus cavernosum with a vasoactive drug combination for vasculogenic and neurogenic erectile dysfunction was performed in 50 men from 10/85-6/87. Acceptance was excellent by all patients and their wives. Transitory hematomas appeared in 6/50 (12%) cases. Local infections, cavernitis or systemic side effects were not observed. In 3/50 (6%) patients prolonged erections occurred. Treatment included drainage of the intracavernous blood, manual decompression and if necessary additional application of 1 mg metaraminol under extreme caution. Only 1 (2%) patient developed a nodular tunica fibrosis at the injection site which disappeared spontaneously upon cessation of treatment within 4 weeks.     

Histopathological changes and nitric oxide synthase activity in corpus cavernosum from rats with neurogenic erectile dysfunction

OBJECTIVE: To investigate changes in histology and nitric oxide synthase (NOS) activity in cavernosal tissues from rats with neurogenic erectile dysfunction induced experimentally. MATERIALS AND METHODS: Twenty-four adult male Sprague-Dawley rats were divided equally into three groups and underwent a sham operation (control, group 1), unilateral (group 2) or bilateral (group 3) cavernosal nerve resection. Three months later they were killed and the cavernosal tissues analysed histologically by light and transmission electron microscopy, with NOS activity detected using an NADPH-diaphorase staining technique. RESULTS: On light and electron microscopy, while penile nerves and cavernosal smooth muscle cells had a normal morphological appearance in the eight control rats, there were degenerative changes of the myelinated penile nerves and axonal fibrosis in groups 2 and 3. However, these changes were not significant. Using NADPH-diaphorase staining, NOS activity was detected in all three groups in endothelial cells and cavernosal structures. However, the staining was more intense in endothelial cells and cavernosal muscles of rats in group 2 than in the other groups. CONCLUSION: NOS activity was increased in the cavernosal tissue after cavernosal denervation, but the pharmacological action of nitric oxide may be impaired.     

Acceptance of corpus cavernosum auto-injection therapy in long-term treatment of erectile dysfunction]

A total of 121 patients suffering from erectile dysfunction were evaluated by means of a questionnaire. In all these cases corpus cavernosum autoinjection therapy had been recommended. The aim of the study was to determine the acceptance and complications of the therapy, general level of satisfaction with it, and reasons for refusal. Completed questionnaire were received from 89 patients (75.5%), 56.2% of whom had applied the therapy continuously, while 18% had broken it off and 25.8% had never started on therapy (cumulative dropout rate 43.8%). The complications encountered were temporary hematomas (25.8) and deviations of the penis (10.1%). In 6.1% indurations of the penis were found. The frequency of prolonged erection needing an antidote was 0.07% in the continuous treatment group. The relatively high drop-out rate in the autoinjection therapy group shows the necessity for intensive patient care and the need for alternative therapy options and a more critical view of reports already published of success with autoinjection therapy.     

Characterization of corpus cavernosum smooth muscle cell phenotype in diabetic rats with erectile dysfunction

Phenotypic modulation from a contractile to a proliferative state within vascular smooth muscle cells has a critical role in the pathogenesis of a variety of cardiovascular diseases. To investigate the characterization of corpus cavernosum smooth muscle cell phenotype in diabetic rats with erectile dysfunction, a group of Sprague-Dawley rats (n=30) were induced by intraperitoneal injection of streptozotocin (60?mg?kg(-1)) and screened by subcutaneous injection of apomorphine (100?μg?kg(-1)) for the measurement and comparison of the penile erections, and then three different groups were defined. Primary corpus cavernosum smooth muscle cells were cultured and passaged. The cavernous tissue segments were subjected to quantitative real-time polymerase chain reaction to determine the expressions of smooth muscle α-actin (SMA), SM myosin heavy chain (SMMHC), smoothelin, calponin and myocardin. Cell contractility in vitro and western blot analysis of SMA and SMMHC in the cavernous tissues and cells were determined. Compared with the control group (n=8) and the diabetes mellitus group (n=5), the expressions of SMA, calponin, SMMHC, smoothelin and myocardin mRNA were decreased in the cavernous tissues in rats of the diabetic erectile dysfunction group (n=15; P=0.001 and 0.02, P=0.014 and 0.012, both P<0.001, P=0.005 and <0.001, P=0.003 and 0.035, respectively). The levels of SMA and SMMHC proteins showed a significant decrease in cavernous tissues and cultured cells in rats of the diabetic erectile dysfunction group. Cells of the diabetic erectile dysfunction group exhibited significantly less contractility compared with those of other groups (P<0.001). Corpus cavernosum SM cell possesses the ability to modulate the phenotype under hyperglycemic conditions, which could have a key role in the pathogenesis of diabetic erectile dysfunction.     

Revascularization of corpus cavernosum for erectile failure.

Revascularization of the penis for erectile disability was performed in 10 patients in whom arterial obstruction had been documented. Inferior epigastric artery anastomosis to the corpus cavernosum was not successful, although bypass grafts from the femoral artery to the corpus cavernosum have helped restore potency in 4 of 6 patients. Complications included embolus to the leg, postoperative hemorrhage, and three small hematomas. This procedure can be considered a clinical experiment and would call for more study to assess its ultimate value.     

The value of corpus cavernosum electromyography in erectile dysfunction: current status and future prospect

In the last decade, several investigators have tried to develop corpus cavernosum electromyography (CC-EMG) as a direct clinical method to evaluate the state of the penile autonomic innervation and the cavernous smooth muscle. Both basic and clinical studies have shown promising results. However, its application as a diagnostic tool with clinical relevance was hindered by insufficient knowledge of cavernous smooth muscle electrophysiology, lack of standardization, technical and practical difficulties and problems in the interpretation of the results. Recently, the European Commission created the so-called COST Action B18 (corpus cavernosum EMG in erectile dysfunction), aiming to strengthen the coordination of the European research groups and give the development of CC-EMG a new impetus. This review presents an overview of the physiological background, the current status of CC-EMG, and discusses possibilities for further developments.