阴茎包埋对海绵体内一氧化氮合酶的影响

目的:探讨阴茎包埋对海绵体内一氧化氮合酶(NOS)活性的影响。方法:通过建立大鼠隐匿阴茎模型获得实验标本,分2个月组、4个月组和6个月组进行观测,每组80只大鼠。各阶段中包括包埋组(n=50)、假手术组(n=15)和正常组(n=15)。称量大鼠体重和海绵体重量后,采用化学比色法检测海绵体内NOS活性。结果:各阶段包埋组阴茎海绵体重量、体重及两者的比值与正常组和假手术组比较均无显著性差异(P>0.05);包埋降低大鼠阴茎海绵体组织的NOS活性(2个月组P>0.05,4个月组P<0.05,6个月组P<0.01)。结论:阴茎包埋可影响海绵体内NOS活性,且与包埋时间呈正相关,但对海绵体外观和重量无明显影响。     

高脂血症大鼠阴茎海绵体中VEGF含量变化对NOS的影响

目的 探讨高脂血症引起大鼠阴茎海绵体中血管内皮生长因子(VEGF)含量的变化,以及这种变化对一氧化氮合酶(NOS)活性的影响.方法 将32只雄性wistar大鼠随机分为2组,高脂血症组和正常对照组各16只,通过建立大鼠高脂血症模型,于实验开始后10d和20d,分别随机取各组1/2大鼠观察其勃起功能;采用免疫组化方法观察大鼠阴茎海绵体内VEGF,并用图像分析法测定其含量;用分光光度法分别测定海绵体匀浆中NOS活性.结果 造模开始后第10天,高脂血症组和正常对照组相比,大鼠阴茎海绵体内VEGF含量、NOS活性及阴茎勃起次数差异均有显著性(P＜0.05):造模开始后第20天,两组的VEGF含量、NOS活性及阴茎勃起次数差异均有显著性(P＜0.05).高脂血症组VEGF含量、NOS活性及阴茎勃起次数,第20天均明显低于第10天,两两比较差异均有显著性(P＜0.05).结论 高脂血症大鼠阴茎海绵体中VEGF明显减少,内皮细胞增殖下降,进而NOS下降,这可能是高脂血症引起勃起功能障碍(ED)的发病机制之一.     

高脂血症对大鼠阴茎海绵体中NOS及CO含量影响的研究

目的 探讨高脂血症对大鼠阴茎海绵体中一氧化氮合成酶(NOS)活性及一氧化碳(CO)浓度的影响及两者之间的关系.方法 将32只雄性wistar大鼠随机分为2组:高脂血症组和正常对照组,每组各16只大鼠.通过建立大鼠高脂血症模型,于实验开始后10d和20d,分别随机取各组1/2大鼠观察其勃起功能;用分光光度法分别测定海绵体匀浆中NOS活性和CO浓度.结果 在第10天及第20天,高脂血症组和正常对照组相比,大鼠阴茎海绵体内NOS活性、CO浓度及阴茎勃起次数差异均有统计学意义(P<0.05);而且第20天高脂血症组NOS活性、CO浓度及阴茎勃起次数,均明显低于第10天,两两比较差异均有统计学意义(P<0.05).结论 高脂血症大鼠阴茎海绵体中NOS及CO下降,这可能是高脂血症引起勃起功能障碍的发病机制之一.     

甲状腺素对大鼠阴茎海绵体内NOS及CO含量影响的研究

目的:建立甲状腺功能亢进及甲状腺功能减退Wistar大鼠动物模型,检测其阴茎海绵体内NOS及内源性一氧化碳(CO)的含量,探讨甲状腺素对大鼠勃起功能的影响及内源性CO在阴茎海绵体勃起过程中的作用,进一步讨论甲状腺素对人类勃起功能的影响。方法:将50只3月龄雄性Wistar大鼠随机均分为甲亢组、甲亢治疗组、甲减组、甲减治疗组及正常对照组。用紫外分光光度计分别测定阴茎海绵体内NOS及CO的含量。结果:无论甲状腺素增多及减少都会使大鼠阴茎海绵体NOS含量降低(P<0.01),并且甲减组阴茎海绵体内NOS活性低于甲亢组(P<0.01)。无论甲状腺素增多还是减少都会使大鼠阴茎海绵体内CO含量降低(P<0.01),并且甲亢组阴茎海绵体内CO活性低于甲减组(P<0.01)。在对甲减组及甲亢组进行治疗后其CO及NOS的含量得到提高,与正常对照组无显著差异(P>0.05)。结论:甲状腺功能紊乱情况下阴茎海绵体中NOS和CO的浓度均减低;甲状腺功能紊乱被及时纠正后阴茎海绵体中CO及NOS的含量可恢复到正常水平。在相同条件下甲状腺功能低下对性功能的损害强于甲状腺功能亢进对勃起功能的损害。     

阴茎包埋对海绵体结构和发育的影响

目的探讨阴茎包埋对海绵体结构和发育的影响。方法通过建立隐匿阴茎动物模型获得实验标本，分2个月组、4个月组和6个月组进行观测。检测海绵体质量和大鼠体重，观察发育情况；Masson染色分析海绵体内平滑肌和纤维结缔组织的含量来了解海绵体结构的变化。结果各阶段包埋组动物阴茎海绵体质量、体重及两者的比值与正常组和假手术组两两比较差异均无统计学意义（P〉0．05）；各阶段中包埋组的海绵体平滑肌面积下降（2个月组P〉0．05，4个月和6个月组P〈0．05），纤维结缔组织面积增加（2个月和4个月组P〉0．05，6个月组P〈0．05）血管窦的面积减少（2个月和4个月组P〉0．05，6个月组P〈0．05），且组织的正常形态发生改变。结论阴茎包埋可影响海绵体内平滑肌和纤维结缔组织的含量及组织排列的正常形态，且与包埋时间正相关，但对海绵体的大体观无明显影响。     

尼古丁对大鼠阴茎海绵体内源性CO浓度及NOS活性的影响

目的:观察尼古丁对成年雄性大鼠阴茎海绵体内源性一氧化碳(CO)浓度及一氧化氮合酶(NOS)活性的影响,探讨吸烟对勃起功能损害的可能机制。方法:40只成年雄性Wistar大鼠分为4组,尼古丁注射1个月组、2个月组、3个月组和对照组,尼古丁注射组尼古丁0.5 mg/(kg.d)皮下分别注射1、2、3个月,对照组注射生理盐水。处理后,取阴茎海绵体,用改良双波长分光光度法检测CO浓度,改良Griess法检测NOS活性。结果:对照组CO浓度为(13.66±0.40)μmol/mg prot,NOS活性为(9.72±0.47)U/mg prot。尼古丁注射1个月,CO浓度和NOS活性分别下降为(12.43±0.56)μmol/mg prot和(8.44±0.69)U/mg prot,显著低于对照组(P均<0.01);尼古丁注射2个月,CO浓度和NOS活性分别下降为(11.41±0.52)μmol/mg prot和(7.53±0.24)U/mg prot,显著低于对照组和尼古丁注射1个月组(P<0.01);尼古丁注射3个月,海绵体CO浓度和NOS活性分别下降为(10.52±0.59)μmol/mg prot和(6.64±0.31)U/mg prot,均显著低于对照组和尼古丁注射1个月、2个月组(P均<0.01)。结论:尼古丁可导致成年雄性大鼠阴茎海绵体内源性CO浓度及NOS活性下降,提示内源性CO及NOS参与吸烟引起勃起功能障碍的病理生理过程。     

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。     

血管活性肠多肽基因转导增强阴茎勃起功能

目的 探讨阴茎海绵体勃起神经递质血管活性肠多肽(VIP)基因转导入糖尿病大鼠阴茎海绵体并表达多量VIP以增强勃起功能的可行性.方法 将构建的重组质粒pcDNA3/VIPcDNA注射入链尿佐菌素诱导的糖尿病大鼠阴茎海绵体,在注射3 d后测定阴茎海绵体内压(ICP),分别以正常空白、空白、单纯注射缓冲液和单纯注射pcDNA3载体对照.Dot blot法测定阴茎组织中VIP mRNA表达水平.结果 糖尿病大鼠阴茎海绵体注射重组质粒后3 d,电刺激海绵体神经,ICP较对照组明显升高(P＜0.05);阴茎组织VIP mRNA表达增多(P＜0.05).结论 成功将重组质粒pcDNA3/VIP cDNA转导入糖尿病阴茎海绵体,VIP mRNA表达增加能增强糖尿病大鼠阴茎海绵体的勃起功能.     

利拉鲁肽对糖尿病大鼠阴茎海绵体eNOS表达的影响

目的:研究胰高血糖素样肽-1(GLP-1)类似物利拉鲁肽对糖尿病(DM)大鼠阴茎海绵体内皮型一氧化氮合酶(e NOS)表达的影响,探讨利拉鲁肽对DM勃起功能障碍(DED)大鼠勃起功能的作用。方法:取6周龄雄性SD大鼠,分为正常对照组(NC,n=10)与实验组(n=20),实验组构建DM大鼠模型,随机将实验组分为DM组(n=8)与GLP-1组(n=8)。干预12周后,检测各组空腹血糖(FPG)、空腹胰岛素(FINS)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、睾酮、白介素-6,计算胰岛素敏感指标Homa-IR与Homa-β,比较各组大鼠勃起功能;Western印迹检测各组大鼠阴茎海绵体组织Akt/p-Akt、e NOS/p-e NOS的表达。结果:DM组大鼠勃起次数(0.90±1.14)及勃起率(37.5%)较GLP-1组(2.90±1.53,25.0%)、NC组(4.20±1.05,100%)均明显减少(P0.05);GLP-1组大鼠勃起率及勃起次数亦少于NC组大鼠(P0.05)。免疫荧光染色提示e NOS主要表达在海绵体血管、血窦内皮细胞的细胞质中,DM组、GLP-1组e NOS蛋白表达水平显著低于正常对照组(P0.05),且GLP-1组明显高于DM组(P0.05)。DM、GLP-1组大鼠阴茎组织e NOS/p-e NOS表达水平较NC组明显下降(P0.01或0.05)。与DM组相比,GLP-1组大鼠阴茎组织p-e NOS表达水平明显升高(P0.05)。3组大鼠阴茎组织Akt比较无显著差异(P0.05)。DM、GLP-1组大鼠阴茎组织p-Akt表达水平较NC组明显下降(P0.01或0.05)。与DM组相比,GLP-1组大鼠阴茎组织p-e NOS表达水平明显升高(P0.05)。结论:GLP-1可能通过调节Akt/e NOS信号通路,保护阴茎海绵体组织内皮细胞功能,改善DED大鼠的勃起功能,提示GLP-1的使用可能是将来治疗和预防DED的重要方法之一。     

阴茎包埋对海绵体形态结构的影响

目的:探讨阴茎包埋对大鼠阴茎海绵体形态结构的影响。方法:通过建立隐匿阴茎大鼠模型获得实验标本,分2月组、4月组进行观测,每组25只大鼠。各阶段又分包埋组(15只)、正常组(10只),光镜和电镜下观察海绵体形态结构的改变。结果:阴茎包埋2月组海绵体形态结构无明显变化,4月组病理改变较为明显,光镜下见大鼠阴茎海绵体平滑肌细胞,内皮细胞分布杂乱,细胞间大量间质组织增生,海绵窦狭窄;电镜下见阴茎海绵体平滑肌细胞及内皮细胞萎缩、线粒体退变、内质网扩张,致密体及收缩纤维减少,脂滴增加,空泡形成。包埋组与正常组阴茎海绵体在外观和重量上无明显差异(P>0.05)。结论:阴茎包埋对海绵体外观和重量无明显影响,但随着包埋时间的延长,海绵体组织发生超微结构上的病理改变。     

衰老对大鼠阴茎勃起机制的影响

为探讨衰老对阴茎勃起机制的影响程度,以不同月龄的雄性大鼠分别观察海绵体神经诱导的阴茎勃起,海绵体平滑肌的舒张功能以及阴茎含ＮＯＳ神经和平滑肌量。结果表明电刺激老年鼠海绵体神经可取得与年轻和中年鼠相当的勃起压力,但勃起潜伏期较长。注射罂粟碱提示老年海绵体平滑肌顺应性较年轻鼠差。检查还提示老年鼠阴茎内含ＮＯＳ神经和平滑肌纤维量有一定减少。实验证实衰老对阴茎勃起机制中关键性的组织结果和功能有一定影响,但     

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。     

The TGF-β1/Smad/CTGF pathway and corpus cavernosum fibrous-muscular alterations in rats with streptozotocin-induced diabetes

Diabetes-associated erectile dysfunction is associated with increased extracellular matrix deposition and reduced smooth muscle content in the corpus cavernosum. The mechanisms of these processes are not well understood. In this study, we investigated fibromuscular changes in the corpus cavernosum of rats with streptozotocin-induced diabetes to determine the mechanisms underlying pathologic changes in penile structure and function. Forty 8-week-old Sprague-Dawley rats were randomly distributed into control and diabetic groups. Diabetes was induced by a one-time intraperitoneal injection of streptozotocin 60 mg/kg. Twelve weeks later, erectile function was measured by cavernous nerve electrostimulation with real-time intracorporal pressure assessment. The penis was harvested for histologic examination (Masson trichrome stain, picrosirius red stain, Hart elastin stain, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling, and immunohistochemistry) and Western blot. Diabetes significantly attenuated erectile response to cavernous nerve electrostimulation. Diabetic animals exhibited a decreased smooth muscle/collagen ratio in the corpus cavernosum. The ratio of collagen I to II fibers was significantly lower in the corpora of diabetic rats compared with controls. Cavernous elastic fibers were fragmented in diabetic rats. There was up-regulation of the transforming growth factor β1/Smad/connective tissue growth factor signaling pathway in diabetic rats. Phospho-Smad2 expression was higher in smooth muscle cells and fibroblasts of diabetic rats, as was the apoptotic index. The up-regulation of the transforming growth factor β1/Smad/connective tissue growth factor signaling pathway might play an important role in diabetes-induced fibrous-muscular structural changes and deterioration of erectile function.     

Nitric oxide synthase gene transfer for erectile dysfunction in a rat model

OBJECTIVE: To determine whether over-expression of nitric oxide synthase (NOS) in the corpus cavernosum of the penis improves erectile function, as NO is an important transmitter for genitourinary tract function, mediating smooth muscle relaxation and being essential for penile erection. MATERIALS AND METHODS: The inducible form of the enzyme NOS (iNOS) was introduced into the corpus cavernosum of adult Sprague-Dawley rats (250-300 g) by injecting a solution of plasmid, adenovirus or adenovirus-transduced myoblast cells (adeno-myoblasts). Plasmid, adenovirus and adeno-myoblasts encoding the expression of the beta-galactosidase reporter gene were also injected into rats. RESULTS: Throughout the corpora cavernosum there was expression of beta-galactosidase after injecting each of the three solutions. Maximum staining was greatest for adeno-myoblast, then adenovirus and then plasmid. The mean (sd) basal intracavernosal pressure (ICP) of iNOS-treated animals (adenovirus and adeno-myoblast) increased to 55 (23) cmH2O, compared with naive animals with a basal ICP of 5 (6) cmH2O (P = 0.001). Stimulating the cavernosal nerve (15 Hz, 1.5 ms, 10-40 V, 1 min) resulted in a doubling of the ICP (adenovirus and adeno-myoblast) from the basal level of the iNOS-treated animals. Direct in situ measurement of NO showed the release of 1-1.3 micro mol/L in the adeno-myoblast penis. CONCLUSION: Myoblast-mediated gene therapy was more successful for delivering iNOS into the corpus cavernosum than direct adenovirus injection or plasmid transfection. Surprisingly, implanting muscle cells into the penis is not only feasible but also beneficial. Gene therapy for NOS may open new avenues of treatment for erectile dysfunction. Control of NOS expression would be necessary to prevent priapism.     

Effects of Icariside II on Improving Erectile Function in Rats With Streptozotocin-Induced Diabetes

Icariin and icariside II (ICA II), 2 active components isolated from herba epimedii, have a closely structural relationship. There is evidence that icariin may be useful in the treatment of erectile dysfunction (ED); however, the study on the therapeutic efficacy of ICA II on ED is currently scant. We investigated the effects of ICA II on improving erectile function of rats with streptozocin-induced diabetes. Fifty 8-week-old Sprague-Dawley rats were randomly distributed into normal control and diabetic groups. Diabetes was induced by a one-time intraperitoneal injection of streptozocin (60 mg/kg). Three days later, the diabetic rats were randomly divided into 4 groups including a saline-treated placebo arm and 3 ICA II-treated models (1, 5, and 10 mg/kg/d). After 3 months, penile hemodynamics was measured by cavernous nerve electrostimulation (CNE) with real time intracorporal pressure assessment. Penises were harvested with subsequent histological examination (picrosirius red stain, Hart elastin stain, and immunohistochemical stain) and Western blots to explore the expression of the nitric oxide-cyclic guanosine monophosphate (NO-cGMP) and transforming growth factor β1 (TGFβ1)/Smad2 signaling pathways. Diabetes significantly attenuated erectile responses to CNE. Diabetic rats had decreased corpus cavernosum smooth muscle/collagen ratio and endothelial cell content relative to the control group. The ratio of collagen I to III was significantly lower in the corpora of diabetic rats; furthermore, cavernous elastic fibers were fragmented in the diabetic animals. Neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase, and vascular endothelial growth factor were expressed at lower levels in the diabetic group; ICA II-treated diabetic rats had higher expression in the penis relative to placebo-treated diabetic animals. Both the TGFβ1/Smad2/connective tissue growth factor (CTGF) signaling pathway and apoptosis were down-regulated in the penis from ICA II-treated rats. ICA II treatment attenuates diabetes-related impairment of penile hemodynamics, likely by increasing smooth muscle, endothelial function, and nNOS expression. ICA II could alter corpus cavernosum fibrous-muscular pathological structure in diabetic rats, which could be regulated by the TGFβ1/Smad2/CTGF and NO-cGMP signaling pathways.     

Expression of oxytocin receptor in diabetic rat penis

Oxytocin receptor (OTR) expressed in the rat penis and mediated the contractility of the corpus cavernosum smooth muscle both in vitro and in vivo, and OTR could maintain penile detumescence; however, the expression of OTR in diabetic rat penis remains unknown. In the present study, we investigated the expression of OTR in diabetic rat penis. The experimental rats were randomly divided into control group and STZ-diabetic rats group. The expressions of mRNA and protein were examined by real-time quantitative PCR, Western blotting and immunohistochemistry respectively. Erectile function was evaluated by measuring intracavernous pressure following electrostimulation of the cavernous nerves. mRNA and protein expression of OTR significantly increased in diabetic rats group compared with the control group. Erectile function of diabetic rats group significantly decreased compared with the control group. Our data showed that the expression of OTR significantly increased in diabetic rats group and OTR may involve in the development of diabetic erectile dysfunction.     

去势对大鼠阴茎海绵体功能和结构的影响

目的 :探讨雄激素对阴茎海绵体功能和结构的影响。　方法 :30只成年雄性大白鼠随机分为 3组 :阉割组、替代组及假手术对照组。于 1周后取阴茎海绵体 ,用紫外分光光度计测定其一氧化氮合酶 (NOS)活性 ,同时用ISEL法检测其细胞凋亡情况。　结果 :阉割组海绵体NOS活性下降 70 %并出现细胞凋亡 (P <0 .0 1) ,睾酮替代能阻止NOS活性降低及凋亡的发生 (P >0 .0 5 )。　结论 :雄激素可通过调节NOS活性及细胞的增殖与凋亡而维持阴茎海绵体的结构与功能。