大鼠盆神经节胆碱乙酰转移酶和一氧化氮合酶阳性神经元对尿道外括约肌的支配

目的 观察大鼠盆神经节(MPG)中胆碱乙酰转移酶(ChAT)、一氧化氮合酶(NOS)阳性神经元对尿道外括约肌(EUS)的支配.方法 24只成年SD大鼠随机分为A组(n=12)和B组(n=12).应用辣根过氧化物酶(HRP)逆行示踪技术结合免疫组织化学和酶组织化学双标技术,观察A组大鼠MPG中ChAT阳性神经元和还原型辅酶Ⅱ(NADPH,NOS标志物)阳性神经元对EUS的支配.B组大鼠MPG内注射麦芽凝集素-辣根过氧化物酶(WGA-HRP)进行顺行神经追踪.结果 A组大鼠HRP标记神经元散在分布于MPG内.HRP标记神经元中43.3%(155/358)为ChAT免疫反应阳性,14.5%(49/339)为NADPH组化反应阳性.B组大鼠EUS内有HRP阳性神经末梢.结论 大鼠盆神经节中的神经元对EUS有直接支配,其神经递质有乙酰胆碱和一氧化氮.     

大鼠盆神经节胆碱乙酰转移酶和一氧化氮合酶阳性神经元对尿道外括约肌的支配

目的 观察大鼠盆神经节(MPG)中胆碱乙酰转移酶(ChAT)、一氧化氮合酶(NOS)阳性神经元对尿道外括约肌(EUS)的支配.方法 24只成年SD大鼠随机分为A组(n=12)和B组(n=12).应用辣根过氧化物酶(HRP)逆行示踪技术结合免疫组织化学和酶组织化学双标技术,观察A组大鼠MPG中ChAT阳性神经元和还原型辅酶Ⅱ(NADPH,NOS标志物)阳性神经元对EUS的支配.B组大鼠MPG内注射麦芽凝集素-辣根过氧化物酶(WGA-HRP)进行顺行神经追踪.结果 A组大鼠HRP标记神经元散在分布于MPG内.HRP标记神经元中43.3%(155/358)为ChAT免疫反应阳性,14.5%(49/339)为NADPH组化反应阳性.B组大鼠EUS内有HRP阳性神经末梢.结论 大鼠盆神经节中的神经元对EUS有直接支配,其神经递质有乙酰胆碱和一氧化氮.     

大鼠盆神经节NOS和VIP阳性神经元对阴茎海绵体的支配

目的：研究大鼠盆神经节(MPG)中一氧化氮合酶(NOS)、血管活性肠肽(VIP)阳性神经元对阴茎海绵体的支配。方法：应用辣根过氧化物酶(HRP)逆行示踪技术结合免疫组织化学和酶组织化学双标技术,观察大鼠MPD中还原型辅酶Ⅱ(NADPFI,NOS标志物)阳性和VIP阳性神经元对阴茎海绵体的支配。结果：HRP阳性标记神经元主要分布于MPG内靠近阴茎神经的区域。HRP阳性标记神经元中72％(210／292)为VIP免疫阳性,83％(268／323)为NADPH组化反应阳性。结论：大鼠盆神经节中NOS、VIP阳性神经元对阴茎海绵体有直接支配,NO和VIP作为神经递质或调质参与了阴茎勃起。     

雄激素对大鼠视前内侧区一氧化氮合酶阳性神经元的影响

目的 探讨雄激素对大鼠下丘脑一氧化氮合酶(NOS)阳性神经元的影响。方法 2个月龄雄性大鼠分成A组(去势组)、B组(假手术组)、C组(去势 睾酮替代组)以及D组(保列治喂饲组)，分别于处理前、处理后2周和2个月观察大鼠30min非接触性阴茎勃起(NCE)的次数，对视前内侧区(MPOA)、视上核(SON)、室旁核(PVN)以及终纹床核行尼克酰胺腺嘌呤二核苷酸黄递酶(NADPH—d)组织化学染色。结果 A组和D组大鼠30min NCE次数明显少于B组和C组。在NADPH组织化学染色中，术后2周各组大鼠脑组织阳性着色差异无显著性，术后2个月，A组与D组MPOA阳性着色明显少于B组和C组，而其他脑区阳性着色在各组间差异仍无显著性。结论 大鼠阴茎勃起功能对雄激素具有依赖性，而雄激素对性反应调节中枢的影响可能与调节MPOA NOS阳性神经元有关；其中5α—双氢睾酮可能是主要的雄激素活性成分。     

糖尿病大鼠盆神经节nNOS表达和阴茎海绵体NOS活性的实验研究

阴茎勃起功能障碍（ED）是糖尿病（DM）常见的并发症之一，国外报道DM性ED发病率为50％～75％，其发病机制尚不清楚。勃起是一种神经控制的血流动力学变化过程，盆神经节控制进入阴茎的血流。本研究通过建立DM大鼠模型，检测盆神经节神经元型一氧化氮合酶（nNOS）的表达和海绵体NOS的活性，以进一步探讨DM性ED的发病机制。     

盆神经节海绵体内移植对双侧海绵体神经损伤后大鼠阴茎勃起功能的影响

目的　研究双侧海绵体神经离断后 ,自体盆神经节海绵体内移植对大鼠阴茎勃起功能的影响。方法　将 2 6只雄性SD大鼠 (3～ 6个月 ,30 0～ 4 0 0g 只 )随机分为 2组 :(1)实验组 (n =16 )行双侧海绵体神经离断及左盆神经节移植入左侧阴茎脚内 ;(2 )对照组 (n =10 )仅离断双侧海绵体神经。1个月及 3个月后阿朴吗啡皮下注射检测勃起功能 ,电刺激左阴茎脚移植区域或右侧盆神经节比较海绵体内压的变化 ,NADPH染色检测海绵体内一氧化氮合酶 (NOS)阳性神经纤维 ,透射电镜观察盆神经节移植后的存活情况。结果　两组大鼠 1个月及 3个月后注射阿朴吗啡后均无勃起反应 ;电刺激左阴茎脚盆神经节移植区域后亦不能导致勃起。 1个月后电刺激两组左阴茎脚后的海绵体内压变化分别为 (9 4 1± 3 2 0 )、(4 16± 2 5 8)cmH2 O(t=4 76 9,P <0 0 5 ) ,3个月后分别为 (13 6 7± 4 18)、(5 0 9± 2 74 )cmH2 O(t=9 18,P <0 0 5 )。两组大鼠左阴茎脚区域的NOS阳性神经纤维数目 1个月后分别为 (2 18 7± 2 4 5 )、(18 0± 3 7) (t=14 77,P <0 0 5 ) ,3个月后分别为 (183 2± 19 7)、(19 0± 3 8) (t =12 18,P <0 0 5 ) ;透射电镜证实了神经节移植后的存活。结论　盆神经节海绵体内移植后可以存     

异丙酚对高血压大鼠胸主动脉内皮型一氧化氮合酶和诱导型一氧化氮合酶表达的影响

目的 评价异丙酚对高血压大鼠胸主动脉内皮型一氧化氮合酶(eNOS)和诱导型一氧化氮合酶(iNOS)表达的影响.方法 SD大鼠,雌雄各半,体重240～ 280 g,采用皮下注射去氧皮质酮的方法制备高血压模型,采用随机数字表法,将64只造模成功的大鼠随机分为4组(n＝16):高血压组(H组)、小剂量异丙酚组(P1组)、中剂量异丙酚组(P2组)和大剂量异丙酚组(P3组).P1组、P2组和P3组分别静脉输注异丙酚20、30、40 mg·kg-1·h-13 h,H组给予等容量生理盐水.分别于给药前、给药1h、3h时记录平均动脉压(MAP).给药3h时处死大鼠,摘眼球法采集血样,硝酸还原酶法测定血清一氧化氮(N0)浓度,取胸主动脉,采用RT-PCR和Western blot法测定eNOS mRNA、iNOS mRNA及其蛋白表达水平.结果 与H组比较,P1组、P2组和P3组给药3h时MAP降低,血清NO浓度升高,主动脉eNOS mRNA及其蛋白表达上调,主动脉iNOS mRNA及其蛋白表达下调,且呈剂量依赖性(P<0.05或0.01).结论 异丙酚降低高血压大鼠血压的机制与下调iNOS表达,上调血管内皮细胞eNOS表达,促进NO释放有关.     

去神经支配对青春期大鼠前列腺的影响及神经元型一氧化氮合酶的表达和意义

目的:探讨去神经对青春期大鼠前列腺生长发育的影响及神经元型一氧化氮合酶(nNOS)在其中可能的作用。方法:20只SD雄性大鼠随机分为假手术组(A组)及手术组(B组),采用显微外科技术建立去盆神经节模型,以假手术组为对照组。5周后取前列腺腹侧叶观察其形态结构的变化,并检测前列腺组织细胞凋亡及nNOS的表达。结果:B组大鼠去神经侧前列腺腹侧叶湿重比A组减少了30.8%(P<0.01),腺体明显减少,细胞高度减低,腺泡萎缩,细胞凋亡指数显著增加(P<0.01),nNOS表达下降(P<0.01)。结论:去神经支配引起前列腺组织细胞凋亡,使青春期大鼠前列腺生长发育受阻,nNOS可能在其中发挥重要的作用。     

脊髓神经元型一氧化氮合酶在大鼠神经病理性痛中的作用

目的 探讨脊髓神经元型一氧化氮合酶(nNOS)在大鼠神经病理性痛中的作用.方法 健康雄性SD大鼠40只,体重220～280 g,采用结扎坐骨神经干的方法建立坐骨神经慢性压迫性损伤(CCI)模型.随机分为4组(n=10),Ⅰ组及Ⅱ组暴露坐骨神经干,分别于术后1 d开始鞘内注射选择性nNOS抑制剂7-NI 60 μg[溶于20%二甲基亚砜(DMSO)]10μl)、20%DMSO 10μl,1次/d,连续6d;Ⅲ组及Ⅳ组制备CCI模型,分别于术后1 d开始鞘内注射7-NI 60μg(溶于20%DMSO 10μl)、20%DMSO 10 μl,1次/d,连续6 d.分别于CCI前1 d、CCI后1、3、5、7 d时测定大鼠机械痛阈和热痛阈.于CCI后7 d,各组分别取5只大鼠,取术侧L_(4～6)背根神经节,分别采用实时定量PCR和Western blot法测定nNOS mRNA及蛋白的表达水平.结果 与Ⅰ组和Ⅱ组比较,T_(1～4)时Ⅲ组和Ⅳ组术侧后肢机械痛阈和热痛阈降低(P＜0.05),背根神经节nNOS蛋白及mRNA的表达上调(P＜0.05);与Ⅲ组比较,T_(1～4)时Ⅳ组机械痛阈和热痛阈降低,背根神经节nNOS蛋白及mRNA的表达上调(P＜0.05).结论 脊髓nNOS参与了大鼠神经病理性痛的形成.     

辛伐他汀预处理对脓毒症大鼠胸主动脉诱导型一氧化氮合酶及内皮型一氧化氮合酶表达的影响

目的 探讨辛伐他汀预处理对脓毒症大鼠胸主动脉诱导型一氧化氮合酶(iNOS)及内皮型一氧化氮合酶( eNOS)表达的影响.方法 清洁级雌性Wistar大鼠80只,4月龄,体重200～250 g,采用随机数字表法,将其随机分为4组:正常对照组(Ⅰ组,n=8)、假手术组(Ⅱ组,n=8)、脓毒症组(Ⅲ组,n=32)和辛伐他汀预处理组(Ⅳ组,n=32).Ⅱ组打开腹腔找到盲肠后还纳腹腔,缝合腹壁切口.Ⅲ组和Ⅳ组采用盲肠结扎穿孔法制备大鼠脓毒症模型.Ⅳ组制备脓毒症模型前经胃管灌注辛伐他汀20 mg/kg,1次/d,连续2周,Ⅰ组～Ⅲ组给予等容量生理盐水.Ⅱ组于假手术后6h处死动物,Ⅲ组和Ⅳ组于盲肠结扎穿孔后3、6、24、48 h分别取8只动物,处死,取胸主动脉,采用Western blot法测定iNOS和eNOS表达水平.结果 与Ⅰ组比较,Ⅱ组iNOS和eNOS表达差异无统计学意义(P＞0.05),Ⅲ组和Ⅳ组iNOS表达上调,eNOS表达下调(P＜0.05);与Ⅲ组比较,Ⅳ组iNOS表达下调,eNOS表达上调(P＜0.05).结论 辛伐他汀预处理可下调脓毒症大鼠血管内皮细胞iNOS表达和上调血管内皮细胞eNOS表达,对血管内皮细胞功能有保护作用.     

The facilitatory effect of duloxetine combined with pelvic floor muscle training on the excitability of urethral sphincter motor neurons

Introduction and hypothesis  Aim of this study was to investigate the excitability of sphincter motor neurons under the influence of pelvic floor muscle training (PFMT) and duloxetine. Due to their mechanisms of action, there might be a synergistic effect of duloxetine and PFMT in regard to the facilitation of spinal reflexes controlling urethral sphincter contractions and hence continence. Methods  In ten healthy female subjects, clitoral electric stimulation (CES) and transcranial magnetic stimulation (TMS) were used to determine individual motor thresholds for external urethral sphincter (EUS) contractions before and after PFMT, duloxetine, and PFMT + duloxetine. Results  PFMT and duloxetine alone significantly decreased the motor thresholds for EUS contractions during CES and TMS. However, the combined treatment reduced the motor threshold for EUS contractions significantly stronger compared to PFMT or duloxetine alone. Conclusions  The results are suggestive for a synergistic facilitatory effect of PFMT and duloxetine on sphincter motor neuron activation.     

人工体神经-内脏神经反射弧建立后大鼠盆神经节-膀胱神经通路和递质研究

目的 观察人丁体神经-内脏神经反射弧建立后大鼠膀胱和盆神经节(MPG)之间的神经通路及神经递质性质.方法 8只成年wistar大鼠在体建立人工体神经-内脏神经反射弧.将辣根过氧化物酶(HRP)注射至实验大鼠的膀胱壁肌层,逆行神经追踪,通过免疫组织化学或组织化学方法显示HRP阳性标记细胞中的胆碱已酰转移酶(ChAT)、酪氨酸羟化酶(TH)、血管活性肠肽(VIP)、P物质(SP)和还原型辅酶Ⅱ(NADPH,NOS标志物).结果 支配膀胱的神经元多位于MPG主体部,呈圆形或椭圆形,无明显突起,多数为ChAT(40.1%,112/279)和TH(20.4%,50/245)免疫阳性,少数为VIP(10.7%,25/233)、NOS(6.5%,15/230)、SP(3.3%,8/242)反应阳性.结论 人工体神经-内脏神经反射弧建立后MPG中支配膀胱的神经元以胆碱能和肾上腺素能为主,VIP、SP和一氧化氮(NO)可能发挥神经调质作用.     

The distribution of neuronal and inducible nitric oxide synthase in urethral stricture formation

PURPOSE: The distribution of neuronal (n) and inducible (i) nitric oxide synthase (NOS) may have a role in the maintenance of normal urethral spongiosum and during the development of spongiofibrosis in urethral stricture disease. MATERIALS AND METHODS: Eight normal and 33 strictured human bulbar urethras were studied by histological and immunohistochemical techniques for the neuronal markers S-100, nNOS and iNOS. The smooth muscle-to-collagen ratio was calculated by morphometric analysis of Masson's trichrome sections. Immunohistochemical staining patterns of the neuronal markers in normal urethral tissue was compared to that in urethral stricture tissue with spongiofibrosis. RESULTS: The smooth muscle-to-collagen ratio was significantly lower in the strictured urethra compared to that in the control group (p = 0.001). In the strictured bulbar urethra nNOS immunoreactivity was decreased compared to that in normal urethral tissue. The severity of spongiofibrosis corresponded to the loss of nNOS immunoreactivity. iNOS immunoreactivity was found in strictured urethral epithelium and spongiosal tissue, whereas the control group was nonimmunoreactive to iNOS. CONCLUSIONS: Urethral stricture formation is a fibrotic process associated with significant changes in NOS metabolism. Abnormal collagen synthesis following urethral trauma may be stimulated by inappropriate iNOS activity. A functional nerve supply to the urethral spongiosum seems to be crucial in the maintenance of the unique ultrastructure of the urethral spongiosum.     

BPH患者尿道外括约肌运动单位电位正常值的检测

目的:测定BPH患者尿道外括约肌运动单位电位(motor unit potential,MUP),统计出相应参数的正常值,为临床上发现"高危"BPH患者尿道外括约肌异常,降低"高危"BPH患者术后尿失禁发生率提供一定依据.方法:采用同心圆针电极经会阴穿刺至尿道外括约肌,用尿动力仪记录肌电信号,测定并记录BPH患者尿道外括约肌MUP.结果:本研究测定BPH患者80例,对每例患者收集符合标准的电位5～10个,记录每个电位的时限和波幅,经统计学分析得出,BPH患者尿道外括约肌MUP的参考值时限为3.46～11.26 ms,平均7.36 ms,波幅为55～607 μV,平均237 μV.结论:本研究得出BPH患者的MUP正常值可以用来评估预测相关"高危"BPH患者的尿道外括约肌可能存在的神经肌肉病变,对避免及分析术后尿失禁的原因具有一定的临床运用价值.     

Computer analysis of the electromyogram of the external urethral sphincter

The electromyograms of canine external urethral sphincter were recorded and analyzed with a microcomputer (Nihon Koden Co., ATAC 450). There was no observable change in the electromyographic activity of the external urethral sphincter in response to an α-blocker (phentolamine) or a β-blocker (propanolol). However, there was complete abolition of electromyographic activity following the administration of neuromuscular blocking agents (suxamethonium or pancuronium). The opinions that the α-adrenerpic system acts predominantly on the external urethral sphincter via the pudendal nerve cannot be supported by the data obtained in this study.     

Nitric oxide synthase in the external urethral sphincter of the sheep: immunohistochemical and functional study

PURPOSE: We studied the distribution of neuronal nitric oxide synthase (nNOS) and the effects of nitric oxide (NO) modulating drugs on contractile function of the external urethral sphincter of lambs. Gender differences were evaluated. MATERIALS AND METHODS: Longitudinal and transverse sections of the external urethral sphincter from 10 female and 10 male lambs were studied using reduced nicotinamide adenine dinucleotide phosphate-diaphorase histochemistry and nNOS immunocytochemistry. Isometric contractile responses to electrical field stimulation were recorded from external urethral sphincter preparations from 47 female and 45 male lambs and the effects of NO modulating drugs were evaluated. RESULTS: We detected nNOS in the sarcolemma of some but not all striated fibers, where nNOS seems to be concentrated at the neuromuscular junction. In addition, nNOS was present in nerve fibers and intramural ganglia. The density of innervation decreased toward the distal part of the external urethral sphincter and was higher in male preparations. No significant functional effects of the NOS inhibitor NG-nitro-L-arginine (10 mM.) or the NO donors diethylamine and spermine NONOate (Sigma Chemical Co., St. Louis, Missouri) (5 mM. each) on external urethral sphincter isometric contractility were found in either gender. CONCLUSIONS: Despite the evidence for nNOS at the sarcolemma and nerve fibers of the external urethral sphincter the physiological relevance of these immunohistochemical findings remains to be determined.     

The innervation of the external urethral sphincter; An ultrastructural study in male human subjects

Summary Innervation of the external urethral sphincter (EUS) was studied in male human subjects. In the region of EUS at the distal end of prostatic urethra, a large axon bundle surrounded by perineurium was evident in the intramural connective tissue gap. Because of the presence of dense core vesicles, the small nonmyelinated axon profiles in the bundle were considered to be adrenergic. After ramifications to smooth musculature, the axons were traced to the EUS. In the EUS, axon bundles containing many nonmyelinated axons were recognized as a sole autonomic nerve among the striated muscle cells. A single or at most two or three axons were surrounded by a Schwann cell, and some possessed dense core vesicles which suggested an adrenergic function. These autonomic adrenergic nerve ends formed surface junctions with the striated muscle of EUS. The clinical relevance of these data are discussed.