衰老对大鼠阴茎海绵体NOS I的表达和NOS活性的影响

目的 :探讨衰老对大鼠阴茎海绵体一氧化氮合酶Ⅰ (NOSⅠ)mRNA、蛋白的表达和NOS活性的影响。　方法 :30只雄性SD大鼠按不同月龄分为成年组、老年组和衰老组 ,应用Western印迹、RT PCR方法分别检测不同年龄组阴茎海绵体NOSⅠ蛋白及mRNA的表达 ;用紫外分光光度计测定不同年龄组阴茎海绵体NOS的活性。　结果 :成年组NOSⅠ 蛋白的表达量最高 ,老年组和衰老组显著降低 ,分别为成年组的 75 .6 %和 6 1.2 % ;NOSⅠmRNA的表达与蛋白表达的变化一致 ;老年组NOS活性与成年组差异无显著性 (P >0 .0 5 ) ,衰老组NOS活性明显降低 ,是成年组的70 .4 % ,并且差异非常显著 (P <0 .0 1)。　结论 :衰老引起NOSⅠ 蛋白及mRNA的表达降低和NOS活性的显著降低 ,可能是老年性阴茎勃起功能障碍的主要机制之一。     

自发性高血压大鼠阴茎海绵体组织nNOS变化的实验研究

目的：研究高血压对大鼠阴茎海绵体组织nNOS神经纤维的影响。方法：自发性高血压大鼠(SHR)和同系WKY鼠各12只，都随机分为4周观察组(6只)和12周观察组(6只)。第4周和第12周末时，观察阴茎勃起功能，并检测阴茎组织nNOS神经纤维数目。结果：4周组和12周组中，SHR和WKY鼠在阴茎勃起次数和阴茎组织nNOS神经纤维数目上，都有显著差异(P＜0．01)。SHR4周组和SHR12周组，在阴茎组织nNOS神经纤维数目上，差异也有显著性(P＜0．05)，阴茎勃起次数和勃起率差异无显著性(P＞0．05)，但有减少的趋势。结论：高血压使阴茎组织中nNOS神经纤维数目减少，这可能是高血压引起勃起功能障碍的发病机理之一。     

慢性肾功能衰竭性勃起功能障碍大鼠阴茎海绵体有效成分的测定

目的:探讨慢性肾功能衰竭(CRF)性勃起功能障碍(ED)的发病机制。方法:应用SD雄性大鼠分两期行5/6肾脏切除术,建立CRF动物模型。将假手术组(NCRF组,n=30)、CRF模型大鼠(CRF组,n=45)分别随机均分为Ⅰ(2周)、Ⅱ(4周)、Ⅲ(6周)3组,并分别于2、4、6周注射阿朴吗啡(APO,80μg/kg)后观察大鼠阴茎勃起情况,筛选CRF性ED模型大鼠;测定阴茎海绵体组织一氧化氮合酶(NOS)的活性,及其组织的M asson染色图像分析。结果:CRF性ED大鼠阴茎海绵体组织NOS活性及平滑肌面积显著降低(P<0.01或P<0.05),胶原纤维略有增加,且上述变化与CRF病程密切相关。海绵窦血管腔无明显变化。结论:CRF严重影响阴茎勃起功能,阴茎海绵体组织NOS活性降低及阴茎海绵体平滑肌面积的减少可能是其重要的发病机制。     

糖尿病大鼠阴茎海绵体组织nNOS神经变化的实验研究

目的 :研究糖尿病对大鼠阴茎海绵体组织nNOS神经纤维的影响。　方法 :34只SD雄性大鼠分为 6周组(17只 )和 8周组 (17只 ) ,其中每组各 11只腹腔注射链脲佐菌素制备糖尿病模型 ,其余各 6只注射柠檬酸缓冲液作为空白对照 ,分别在注射 6周和 8周后 ,观察大鼠阴茎勃起功能 ,并采用免疫组化SP法检测糖尿病大鼠阴茎组织nNOS神经纤维的数量。　结果 :2组糖尿病大鼠的阴茎勃起率分别为 37.5 %和 14 .3% ,差异有显著性 (P <0 .0 5 ) ,均明显低于对照组 (P <0 .0 1)。 2组糖尿病大鼠nNOS神经纤维的表达明显低于对照组 (P <0 .0 1)。 8周组中糖尿病大鼠阴茎的勃起功能和nNOS神经纤维的数量明显低于 6周组 ,分别为 (37.6 0± 6 .76 )条和 (2 8.0 0±5 .2 9)条 ,即随着病程的延长 ,勃起功能和nNOS神经纤维的表达下降 (P <0 .0 5 ) ,而对照组大鼠nNOS神经纤维的数量差异无显著性 [(83.0 0± 3.2 2 )vs(81.0 0± 3.6 1) ]。　结论 :糖尿病严重影响勃起功能和nNOS神经纤维的表达。糖尿病大鼠阴茎海绵体组织中nNOS神经纤维的数量明显减少 ,且与病程正相关 ,这可能是糖尿病性勃起功能障碍的发病机理之一。     

乙醇对大鼠性功能影响的实验研究

目的:探讨乙醇对大鼠性功能的影响。方法:将60只Wistar雄性大鼠随机分为对照组、10%乙醇剂量组、20%乙醇剂量组、30%乙醇剂量组和40%乙醇剂量组,每组12只。分别以生理盐水、10%、20%、30%、40%食用乙醇各2ml给大鼠灌胃,每日1次。3个月后,观察各组大鼠性行为和药物诱发阴茎勃起的变化;用全自动化学发光分析仪测定各组大鼠血清睾酮的含量;以80μg/kg体重的阿朴吗啡大鼠颈部注射诱发阴茎勃起;用化学比色法测定阴茎平滑肌中一氧化氮合酶(NOS)的总量。结果:10%和20%乙醇剂量组大鼠药物诱发阴茎勃起的次数没有受到乙醇影响(P>0.05),但10%乙醇剂量组大鼠的乘骑和插入潜伏期延长,20%乙醇剂量组大鼠除了射精潜伏期外,其余性行为参数也受到抑制(P<0.05)。而30%乙醇剂量组与40%乙醇剂量组大鼠的性行为表现、药物诱发阴茎勃起实验均明显受到抑制(P<0.05)。与对照组相比,实验组血清睾酮含量和阴茎组织中NOS活性均减低(P<0.05)。结论:大鼠长期乙醇摄入后性功能障碍的发生与剂量有关。     

2型糖尿病大鼠血浆同型半胱氨酸与阴茎海绵体内NOS和内源性CO的相关性研究

目的:研究2型糖尿病性大鼠血浆同型半胱氨酸(Hcy)与阴茎海绵体内NOS和内源性CO的相关性。方法:选取3月龄雄性Wistar大鼠50只,随机选取10只为对照组(A组);高糖高脂饲料饲养4周后从其他40只大鼠中筛选出30只构建成功的糖尿病(DM)大鼠模型,随机分成3组:DM大鼠组(B组);胰岛素治疗组(C组)和叶酸+维生素B12治疗组(D组)。8周及12周后注射阿朴吗啡观察各组大鼠阴茎勃起情况。12周后测各组大鼠血浆总Hcy含量及阴茎海绵体内NOS活性和CO含量。结果:与A组比较,B组大鼠血浆Hcy浓度明显升高,阴茎勃起功能明显降低,阴茎海绵体NOS活性和CO含量均下降,差异有显著性(P<0.01)。2型DM大鼠中高Hcy血症发生率为55%。与B组比较,C组和D组中大鼠血浆Hcy浓度显著下降,阴茎勃起功能、阴茎海绵体NOS活性均升高(P<0.01),Hcy与NOS(rA=-0.89,rB=-0.76,rC=-0.91,rD=-0.91)及CO含量(rA=-0.82,rB=-0.77,rC=-0.93,rD=-0.81)均呈负相关。结论:2型DM大鼠血浆中的高Hcy可能是引起阴茎海绵体NOS活性下降、CO含量下降,进而导致DM ED发病的分子机制之一。胰岛素、叶酸和维生素B12可以改善DM大鼠的勃起功能,提高阴茎海绵体NOS活性和CO含量。     

大鼠阴茎组织中NOS表达及增龄的影响

为探讨大鼠阴茎组织中一氧化氮合酶分布及增龄对其的影响。本文采用ＮＡＤＰＨｄ组化对不同月龄大鼠阴茎组织进行染色。结果显示ＮＯＳ主要分布于大鼠阴茎组织平滑肌细胞,而内皮和社会纤维含量较少;随年龄增加,阴茎组织中ＮＯＳ表达逐渐减弱,各月龄组间差别明显。     

糖尿病性勃起功能障碍大鼠阴茎海绵体组织中神经生长因子的表达及其治疗作用的研究

目的:本研究通过检测糖尿病性勃起功能障碍(ED)大鼠阴茎组织中神经生长因子(NGF)表达,并使用hNGF进行治疗,以探讨糖尿病性ED发病机制及NGF治疗作用的机制。方法:成年雄性SD大鼠60只,随机取50只大鼠用于制作糖尿病模型,饲养8周后,取正常组和糖尿病组大鼠阴茎海绵体组织,采用RT-PCR和W estern印迹法检测NGF的mRNA及蛋白水平。从造模成功的糖尿病大鼠中筛选出有ED大鼠,把所有大鼠分为5组:正常组、糖尿病性ED组、糖尿病性ED单用NGF组(NGF组)、糖尿病性ED单用胰岛素组(R I组)、糖尿病性ED联合应用NGF和胰岛素组(NGF+R I组,胰岛素通过颈部皮下注射给药,NGF通过腹腔内注射给药),8周后测海绵体内压(ICP),并取所有大鼠阴茎海绵体组织用免疫组化法观察nNOS神经纤维的变化。结果:与正常组相比,糖尿病性ED组大鼠阴茎海绵体组织中NGF的mRNA表达增加,蛋白含量增加。与糖尿病性ED组相比,NGF组、R I组、NGF+R I组ICP水平显著升高(P<0.05);NGF组、R I组、NGF+R I组阴茎组织中nNOS神经纤维水平显著升高(P<0.05)。结论:糖尿病晚期勃起神经出现损伤并发生ED,推测可能与NGF分泌增加的幅度小于高血糖状态对勃起神经的损伤程度有关,也可能与NGF与其相应受体结合转运能力损害有关,给予外源性NGF可能有助于糖尿病性ED局部神经病变减轻和勃起功能改善。提示NGF的异常在糖尿病性ED的发病及治疗中可能具有重要作用。     

血红素氧合酶2在去势大鼠阴茎海绵体内的表达

目的:研究去势大鼠阴茎海绵体血红素氧合酶2(HO-2)和内皮型一氧化氮合酶(eNOS)的表达,探讨雄激素与HO-2、eNOS在ED中的作用及相关性。方法:10周龄雄性SD大鼠40只,分为4、8、12周组和正常对照组各10只,实验组采取手术切除双侧睾丸,对照组采取假手术。分别于术后4、8、12周测定大鼠血清睾酮(T)、阴茎海绵体内压(ICP)、平均颈动脉压(MAP),取阴茎标本,采用Western印迹分析阴茎海绵体HO-2含量,免疫组化分析HO-2和eNOS的表达。结果:去势各组血清T水平较正常对照组显著下降(P<0.05)。经3V、5V电压刺激后去势各组ICP/MAP值明显下降(P<0.05)。HO-2在正常和去势大鼠阴茎海绵体组织均有表达,去势4周组HO-2光密度分布曲线下面积(341.50±99.70)较正常组(876±443.36)和去势8周组(705.00±152.74)明显下降(P<0.05),去势8周与正常组之间无显著变化(P>0.05),去势12周没有检测到HO-2的表达。eNOS主要表达于阴茎海绵体血管内皮细胞,去势组eNOS(123.94±30.23)较正常组(421.21±125.12)差异有显著性(P<0.05)。T与eNOS和HO-2表达呈高度正相关(r=0.976、0.946,P均<0.05)。结论:雄激素可能通过影响大鼠阴茎海绵体HO-2、eNOS的表达参与阴茎勃起功能调控。     

胰岛素对糖尿病大鼠阴茎内nNOS神经纤维的影响

目的探讨糖尿病性阴茎勃起功能障碍（ED）的发病机制及胰岛素的治疗作用。方法注射链脲佐菌素建立糖尿病（DM）大鼠模型，胰岛素治疗组于成模后注射胰岛素。7周和12周后注射阿扑吗啡（APO）进行大鼠阴茎勃起功能实验，取大鼠阴茎和血浆，用ABC免疫组织化学法观察nNOS神经纤维的变化。测定血浆NOS活性。结果（1）与对照组相比，DM组大鼠阴茎勃起次数明显减少;胰岛素治疗后症状缓解;（2）与对照组相比，DM组血浆NOS活性明显增高;DM组血浆NOS活性与病程延长呈负相关;与DM组比较，胰岛素治疗组血浆NOS活性明显降低;（3）与对照组相比，DM组阴茎内nNOS阳性神经纤维明显减少;与DM组比较，胰岛素治疗组nNOS阳性神经纤维表达增加。结论糖尿病性ED阴茎内nNOS阳性纤维的数量及光密度随DM病程的延长而下降;早期给予胰岛素治疗可预防糖尿病大鼠ED的出现及阴茎内nNOS含量的下降。     

Abnormal lipid metabolism down‐regulates adenosine triphosphate synthase β‐subunit protein expression in corpus cavernosum smooth muscle in vitro and in vivo

Metabolic syndrome is closely related to erectile dysfunction (ED), and hyperlipidaemia is considered a major risk factor for ED. Adenosine triphosphate (ATP) synthase is believed to play an important role in metabolic syndrome; it has been hypothesised that ATP synthase contributes to ED development. We have verified this hypothesis using primary cultured human corpus cavernosum smooth muscle (HCCSM) cells treated with excessive free fat acid (FFA) and a high‐fat diet (HFD) mouse model. Our results showed that high fatty factors could cause lipid accumulation in HCCSM cells, which could result in abnormal lipid metabolism, such as high levels of triglycerides, cholesterol and glucose in the HFD mice. There was a remarkable down‐regulation of ATP synthase and p‐Akt after in vivo and in vitro excessive FFA treatments. These results indicated that abnormal lipid metabolism could induce ATP synthase down‐regulation via the Akt phosphorylation pathway and that ATP synthase may be a target of lipotoxicity in corpus cavernosum smooth muscle cells.     

Effects of Korean ginseng berry extract (GB0710) on penile erection: evidence from in vitro and in vivo studies

Several reports have promoted the root-derived Korean red ginseng (KRG; Panax ginseng) as alternative treatment for erectile dysfunction (ED), and ginsenosides are known to be the principal active ingredients of ginseng. Recent studies showed that ginseng berries produce more ginsenosides than KRG; thus, we investigated the ability of the Korean ginseng berry extract GB0710 to relax the penile corpus cavernosum smooth muscle (CCSM) in this study. As a comparative control, the results were compared to those obtained using KRG. In addition, possible mechanisms of action for GB0710 were investigated. While KRG and GB0710 both displayed dose-dependent relaxation effects on precontracted rabbit CCSM in vitro, GB0710 was shown to be more potent than KRG. The GB0710-induced relaxation could be partially reduced by removing the endothelium. In addition, pre-treatment with several nitric oxide (NO) inhibitors significantly inhibited the relaxation of muscle strips. Furthermore, administration of GB0710 increased intracavernosal pressure (ICP) in a rat in vivo model in both a dose- and duration-dependent manner. Intracellular NO production in human microvascular endothelial cells could be induced by GB0710 and inhibited by N^G-monomethyl-L-arginine. In conclusion, GB0710 had a greater relaxation effect on rabbit CCSM than did KRG extract, and increased ICP in a rat model in both a dose- and a duration-dependent manner. This relaxing effect might be mediated by NO production.     

Application of correlation techniques in the analysis of corpus cavernosum electromyographic signals

Aim： To establish an objective, easy-to-use and comprehensive method to analyze corpus cavernosum electromyo- graphic signals （CC-potentials）. Methods： CC-potentials were recorded during flaccidity in 23 young healthy volunteers, with surface electrodes placed on the penile shaft bilaterally. Based on the correlation function of Matlab software, an application program for the analysis of CC-potentials was developed. Individual CC-potentials and their autocorrelation function were evaluated, yielding parameters amplitude （A）, duration （D）, and dominant frequency （DF）. The crosscorrelation function of both longitudinal and bilateral pairs of adjacent electrodes was calculated to assess the similar- ity and mutual delay of CC-potentials recorded simultaneously from different parts of the CC. The parameters derived were squared maximum cross-correlation coefficient （Rmax） and delay （τ）. Based on the absolute value of τ and the corresponding inter-electrode distance, propagation velocity （PV） was calculated. Results. The values of the parameters were determined automatically. No significant difference related to the locations of the electrodes for parameters A, D, and DF was detected. The cross-correlation showed that both longitudinal and bilateral CC-potential pairs had highly similar waveforms （the absolute values of Rmax were 0.80 ± 0.05 and 0.87 ± 0.06, respectively）. PV of longitudinal pairs was estimated as 6.15 ± 3.98 cm/s. Conclusion： The application program for correlation analysis of CC-potentials is a comprehensive and versatile method to analyze corpus cavernosum electromyographic recordings. Its objectiveness makes multi-center application possible.     

生长激素对髂内动脉结扎大鼠勃起功能和nNOS神经纤维的影响

目的 :研究生长激素 (GH)对髂内动脉结扎大鼠勃起功能和神经型一氧化氮合酶 (nNOS)神经纤维的影响。方法 :36只成年雄性Wistar大鼠随机均分为 3组 :假手术组、髂内动脉结扎组和GH干预组。于术后 4周末、8周末 ,分别取各组 1/ 2大鼠 ,观察大鼠勃起功能 ,并用免疫组化SP法检测阴茎组织中nNOS神经纤维的数目。　结果 :4周末 ,GH干预组和髂内动脉结扎组在勃起次数和nNOS神经纤维数目上与假手术组相比差异有显著性 (P<0 .0 5 ) ,前两组之间差异无显著性 (P >0 .0 5 ) ,而勃起率 3组之间差异不明显 (P >0 .0 5 )。 8周末 ,3组在勃起次数和勃起率上差异均不明显 ;而阴茎组织中nNOS神经纤维数目 ,GH干预组高于髂内动脉结扎组 (P <0 .0 0 1) ,而与假手术组比较差异无显著性 (P >0 .0 5 )。　结论 :GH可以改善髂内动脉结扎大鼠勃起功能 ,这种作用可能与其增加大鼠阴茎海绵体nNOS神经纤维数目有关     

糖尿病性勃起功能障碍大鼠阴茎组织内Cx43和eNOS表达的研究

目的:探讨糖尿病(DM)性勃起功能障碍(DMED)与阴茎组织中连接蛋白(Cx43)和eNOS水平变化的相关性。方法:25只SD大鼠腹腔内注射四氧嘧啶建立DM模型,8周后注射阿朴吗啡观察大鼠阴茎勃起情况,筛选出DMED大鼠,应用RT-PCR和免疫组化法分别检测阴茎组织中Cx43和eNOSmRNA表达水平。结果:DMED大鼠阴茎海绵体组织eNOSmRNA表达(0.155±0.157)与DM组(0.508±0.242)比较,显著降低(P<0.01),而Cx43mRNA则显著升高(0.993±0.157vs0.545±0.138,P<0.01),二者存在负相关性(r=-0.987)。免疫组化检测也显示DMED组大鼠阴茎组织平滑肌细胞中eNOS蛋白表达减少,而Cx43蛋白水平增多。结论:阴茎组织内eNOS水平下降可能是DM患者发生ED的主要机制之一;同时,Cx43的表达呈代偿性增加,其功能有待进一步探讨。     

不同剂量香烟烟雾提取物对阴茎海绵体NOS活性和Cx43蛋白的影响

目的:探讨不同剂量的香烟烟雾提取物对雄性大鼠勃起功能的影响,进一步研究吸烟导致ED的发病机制。方法:75只健康雄性SD大鼠(8周龄),随机分为5组(15只/组)。A组为对照组;B组为皮下注射二甲基亚砜(DMSO)组;C组为皮下注射香烟烟雾提取物(CSE)低剂量组;D组为皮下注射CSE中剂量组;E组为皮下注射CSE高剂量组。应用SD雄性大鼠建立皮下注射CSE模型60 d后,经皮下注射阿朴吗啡(APO)后观察大鼠阴茎勃起情况,处死大鼠取阴茎海绵体组织。一部分标本采用激光共聚焦扫描显微镜方法检测缝隙连接蛋白43(Cx43)的表达;另一部分标本采用比色法测定大鼠阴茎海绵体组织中NOS活性。结果:不同剂量CSE组阴茎勃起次数、阴茎海绵体组织NOS活性和海绵体平滑肌中Cx43表达与DMSO组和对照组比较均明显减少(P<0.05)。实验组中,阴茎勃起次数、阴茎海绵体组织NOS活性和Cx43表达均随着CSE剂量的增加而减少。DMSO组与对照组比较差异无统计学意义(P>0.05)。结论:CSE使阴茎海绵体组织NOS活性明显降低、海绵体平滑肌中Cx43蛋白表达明显减少并且严重影响阴茎勃起功能,并且CSE剂量越大,其影响越明显。提示,Cx43蛋白表达下调、NOS活性降低可能是CSE导致ED的发病机制之一。     

Effect of sodium nitropmsside on hemodynamics of corpus cavemosum in Chinese

Aim: To study the effect of sodium nitroprusside (SNP) on the hemodynamics of corpus cavemosum in Chinese men with erectile dysfunction (ED). Methods: In 68 ED patients receiving intracavemous injection (ICI) of SNP, the cavemous hemodynamics were studied by Doppler ultrasonography. Results: The peak flow velocity (PFV), the artery diameter (Ad), the mean velocity of arterial blood (MV) and the vein diameter (Vd) were significantly higher after ICI of SNP than before ICI, but the end diastolic velocity (EDV) did not change significantly. Conclusion: The increase in Vd after SNP suggests that the venous outflow is not invariably decreased during penile election.