豚鼠Oddi括约肌内Cajal样细胞及NOS表达阳性神经元的分布

目的研究Cajal样细胞和一氧化氮合酶(nitric oxide synthase,NOS)表达阳性神经元在成年豚鼠Oddi括约肌(sphincter of Oddi,SO)的分布。方法成年豚鼠SO冷冻切片,c-Kit免疫细胞化学和NADPH-黄递酶组织化学双重染色。结果SO横切面环行平滑肌层内可见少量c-Kit阳性细胞,胞体呈梭形,两端伸出细长的突起。纵切面SO的两个壁分别为:外侧壁和十二指肠壁内SO壁,前者与肠壁结构类似,在深肌丛和肌间神经丛周围可见较多的c-Kit阳性细胞;而十二指肠壁内SO壁的肌层内存在大量c-Kit阳性细胞和突起,其形态与肠壁肌层内Cajal细胞相似。NADPH-黄递酶组织化学染色可见NOS表达阳性神经元广泛分布于SO的肌间神经丛和平滑肌内。c-Kit免疫细胞化学和NADPH-黄递酶组织化学双重染色虽未发现二者的共存,但可见c-Kit阳性细胞及突起存在于NOS表达阳性神经元的附近。结论豚鼠SO内存在的Cajal样细胞可能参与SO自主节律性运动的调控,可能是NOS表达阳性神经元对SO运动发挥调节作用的靶细胞。     

慢传输型便秘大鼠结肠肌间神经丛内VIP能神经、SP能神经的免疫组化研究

目的　探讨结肠肌间神经丛血管活性肠肽 (VIP)能神经及P物质 (SP)能神经在慢传输型便秘发病中的作用。方法　建立大鼠慢传输型便秘模型 ,利用铺片技术制作结肠肌间神经丛标本 ,采用免疫组织化学法在铺片上显示便秘大鼠结肠肌间神经丛内VIP能神经及SP能神经的分布及形态学改变。结果　与正常大鼠比较 ,便秘大鼠结肠肌间神经丛VIP能神经节及节间束细小 ,节内神经元数量明显减少 ,节间束内神经纤维分布稀疏 (平均面密度值 2 3.0 8± 1.82vs 19.35± 1 13,P <0 0 1) ;便秘大鼠结肠肌间神经丛SP能神经节明显增大 ,神经束增粗 ,神经元和神经纤维明显增多 (平均面密度值 15.4 5± 1 0 5vs18.14±1 18,P <0 .0 1)。结论　慢传输型便秘大鼠结肠壁内存在明显的神经病理学改变 ,结肠传输功能障碍可能与结肠肌间神经丛VIP和SP能神经病理改变和 或功能障碍有关。     

烧伤大鼠小肠组织中两型一氧化氮合酶变化的观察

目的 研究烧伤后大鼠小肠组织中两型一氧化氮合酶的变化及与一氧化氮(NO)的关系。方法 采用30％TB-SAⅢ度烧伤大鼠模型,分别检测了小肠组织中NO含量及原生型NOS(cNOS)和诱导型NOS(iNOS)的活性,并分析了NO的变化规律及其与两型NOS之间的关系。结果 烧伤后小肠组中iNOS活性大幅上升,与NO的变化趋势一致,二者呈显著正相关(P<0.01),而cNOS活性呈下降趋势,与NO相关不显著(P<0.05)。结论 烧伤后小肠组织中NO的变化主要受iN-OS活性影响,iNOS活性上升,cNOS活性下降可能与烧伤后胃肠道病理变化密切相关。     

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。     

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.     

吸入麻醉药诱导c—fos基因和一氧化氮合酶在大鼠中枢神经系统的共同表达

目的　本研究旨在探索吸入麻醉药诱导c fos基因和一氧化氮合酶 (NOS)在中枢神经系统共同表达的现象。方法　用免疫组化双标方法 ,在脑切片上观察c fos基因和NOS阳性神经元的分布。结果　吸入麻醉药诱导Fos蛋白在中枢的表达部位及阳性神经元的密度与以往相同。同一实验条件下安氟醚、异氟醚吸入麻醉组的NOS表达明显少于对照组 (P <0 0 5或 0 0 1) ,无一例c fos与NOS在同一神经元内共同表达的现象。结论　安氟醚和异氟醚吸入麻醉作用与抑制在体大鼠脑内NOS的活性有关。     

大鼠睾丸内一氧化氮合酶的表达

目的 :阐明一氧化氮合酶 (NOS)在睾丸内的表达 ,探讨一氧化氮 (NO)在睾丸生殖功能中的作用。　方法 :取雄性SD大鼠睾丸于 4 %多聚甲醛中固定 ,常规石蜡切片。用免疫组化ABC法检测成年大鼠睾丸NOS的表达和定位。　结果 :内皮型NOS(eNOS)、神经型NOS(nNOS)、诱导型NOS(iNOS)在睾丸间质细胞内均有表达 ,精曲小管周围肌样细胞、血管内皮细胞和平滑肌细胞内仅为eNOS免疫反应阳性 ,而血管外膜纤维内仅有nNOS表达。免疫反应物质主要定位于细胞质 ,胞核为阴性。在生精细胞内 3种NOS免疫反应均为阴性。　结论 :3种NOS在睾丸内均有表达 ,不同的NOS分布部位有一定区别     

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.     

早期肠道喂养对烧伤大鼠肠道一氧化氮合酶的影响

目的阐明烧伤大鼠经早期肠道喂养后肠组织中一氧化氮合酶(NOS)的变化规律及其与肠粘膜血流量(IMBF)的关系。方法采用30％TBSAⅢ度烧伤大鼠模型,分为正常对照(C)组,单纯烧伤(B)组和早期喂养(EF)组,分别检测伤前及伤后0,3,6,12,24,48小时肠组织中 NOS 活性,包括原生型 NOS(cNOS)和诱导型 NOS(iNOS),并测定肠粘膜血流量。结果烧伤后各组 cNOS 和 IMBF 呈下降趋势,二者呈显著正相关(r=0.97,P<0.01),而 NOS 总活性和 iNOS 活性都呈上升趋势,IMBF 与 iN-OS 和总 NOS 相关不显著。烧伤后 EF 组 cNOS 和 IMBF 明显高于 B 组,iNOS 则低于 B 组,NOS 总活力两组无显著差异。结论①两型 NOS 中 cNOS 与 IMBF 的关系更加密切。②早期肠道喂养改善烧伤后肠道缺血状况,可能与食物对肠壁神经的刺激从而提高 cNOS 活性有关。     

Involvement of endogenous nitric oxide and c-kit-expressing cells in chronic intestinal pseudo-obstruction

BACKGROUND/PURPOSE: Chronic intestinal pseudo-obstruction (CIP) in infants and children is a motility disorder without apparent mechanical cause. Nitric oxide (NO), an inhibitory neurotransmitter and c-kit cells, essential for the intestinal pacemaker activity, both play a key role in the intestinal motility function. In the current study, the authors investigated the distributive change in the intestinal nitric oxide synthase (NOS) and c-kit cells of patients with CIP. METHODS: Tissues were obtained from 4 patients undergoing bowel resection or biopsy for CIP at laparotomy. For controls, the intestinal specimens were obtained from 4 age-matched cases of intestinal stricture, intussusception, and autopsy with no evidence of gastrointestinal disease. Immunohistochemical studies were performed on paraffin-embedded tissue cross sections with neuronal NOS and inducible NOS monoclonal antibody as well as a rabbit polyclonal antibody against the human c-kit receptor. RESULTS: Under immunohistochemical staining, a greatly increased density of neuronal NOS immunoreactivity and an evidently increased number of intense NOS immunoreactive nerve fibers were observed in the myenteric plexus and circular muscle layers compared with the control sections. In the submucosal plexus and longitudinal muscle layer, there was no change in NOS immunoreactivity. Inducible NOS immunoreactivity was not detected in the control cases. However, in tissues of CIP, almost all the epithelial cells were positively and strongly labeled for inducible NOS immunoreactivity. For c-kit cells staining, the number of c-kit-positive cells in the myenteric plexus and circular muscle layers were greatly less than that in the controls, especially in the myenteric plexus region. CONCLUSION: These findings suggest that sustained production of NO by an increased NOS activity and a deficiency of c-kit cells in the intestine may be related to the pathogenesis of CIP.     

早期肠道喂养对烧伤大鼠肠道一氧化氮合酶的影响

目的阐明烧伤大鼠经早期肠道喂养后肠组织中一氧化氮合酶（ＮＯＳ）的变化规律及其与肠粘膜血流量（ＩＭＢＦ）的关系。方法采用３０％ＴＢＳＡⅢ度烧伤大鼠模型，分为正常对照（Ｃ）组，单纯烧伤（Ｂ）组和早期喂养（ＥＦ）组，分别检测伤前及伤后０，３，６，１２，２４，４８小时肠组织中ＮＯＳ活性，包括原生型ＮＯＳ（ｃＮＯＳ）和诱导型ＮＯＳ（ｉＮＯＳ），并测定肠粘膜血流量。结果烧伤后各组ｃＮＯＳ和ＩＭＢＦ呈下降趋势，二者呈显著正相关（ｒ＝０．９７，Ｐ＜０．０１），而ＮＯＳ总活性和ｉＮＯＳ活性都呈上升趋势，ＩＭＢＦ与ｉＮ－ＯＳ和总ＮＯＳ相关不显著。烧伤后ＥＦ组ｃＮＯＳ和ＩＭＢＦ明显高于Ｂ组，ｉＮＯＳ则低于Ｂ组，ＮＯＳ总活力两组无显著差异。结论①两型ＮＯＳ中ｃＮＯＳ与ＩＭＢＦ的关系更加密切。②早期肠道喂养改善烧伤后肠道缺血状况，可能与食物对肠壁神经的刺激从而提高ｃＮＯＳ活性有关。     

早期肠道喂养对烧伤大鼠肠道一氧化氮合酶的影响

OBJECTIVE: Our previous studies have proved that early enteral feeding could improve intestine blood flow after burn injury. But the mechanism was far from being clarified. This study was attempted to explore the effects of early enteral feeding on nitric oxide synthase (NOS) activity in burned rat small intestine and the relationship between the intestine mucosa blood flow (IMBF) and the activity of NOS. METHODS: The rats were randomly divided into three groups: burned control (B), burned and early enteral feeding (EF), and normal control (C). The activity of NOS including constitute NOS(cNOS) and inducible NOS(iNOS), and the IMBF were determined at postburn 0, 3, 6, 12, 24, 48 hours. RESULTS: It was found that cNOS activity and IMBF were decreased markedly postburn, and there was positive correlation between cNOS and IMBF (r = 0.97, P < 0.01). But the activity of iNOS, total NOS were increased significantly postburn, they had no correlation to the IMBF. In EF group cNOS activity and the IMBF were significantly higher, the iNOS was obviously lower than that of B group and there was no significant difference of total activity of NOS between two groups. CONCLUSION: Our results suggested that NOS which is catalyzed from cNOS may play main role in adjusting IMBF. By using early enteral feeding the activity of cNOS is increased and the ischemic state in small intestine is improved after burn injury.     

氨基胍对饥饿大鼠小肠肌间神经丛神经元诱导型一氧化氮合酶表达的影响

目的 评价氨基胍对饥饿大鼠小肠肌间神经丛神经元诱导型一氧化氮合酶(iNOS)表达的影响.方法 雄性SD大鼠90只,体重230～270 g,采用随机数字表法,将其随机分为3组:正常对照组(C组,n=10)、饥饿组(S组,n=40)和氨基胍+饥饿组(A组,n=40),S组和A组无饲料供应,自由摄水,A组腹腔注射氨基胍150 mg·kg-1·d-1.S组和A组分别于饥饿3、5、7、9d时取10只动物,采用葡聚糖蓝染色法测定小肠传输比率,然后处死动物,取小肠组织,采用NADPH-黄递酶组织化学染色法测定回肠肌间神经丛神经元iNOS表达.结果 与C组比较,S组和A组各时间点小肠传输比率降低,回肠肌间神经丛神经元iNOS表达上调(P＜0.05或0.01);与S组比较,A组于饥饿3、5、7d时小肠传输比率升高,回肠肌间神经丛神经元iNOS表达下调(P＜0.05或0.01).结论 氨基胍改善饥饿大鼠小肠运动功能的机制与下调小肠肌间神经丛神经元iNOS表达有关.     

烫伤大鼠肠道P物质含量和P物质能神经的观察分析

以30%TBSAⅢ度烫伤大鼠为模型,采用 RIA 和 IHC 的方法,动态观察了伤后72小时大鼠空肠免疫活性 P 物质(iSP)的含量及绒毛内 P 物质—免疫反应(SP—IR)阳性神经纤维的变化。结果发现:①烫伤大鼠空肠内 iSP 伤后1小时出现显著升高,4小时仍处于高水平状态;8小时后明显下降,伤后72小时一直处于低水平状态。③绒毛内 SP—IR 阳性神经纤维在形态、分布密度和纤维内 SP—IR 物质的含量都有明显的改变。图像分析定量结果表明:绒毛内 SP—IR 阳性神经纤维分布密度,伤后1小时明显增加,随后减少,48小时再次出现增加;神经纤维内 SP—IR 阳性物质伤后1小时及4小时明显升高,12小时明显下降,24小时后再次升高。实验结果说明:烫伤后大鼠空肠内 P 物质出现应激性释放与消耗,P 物质可能参与了烫伤后大鼠的肠道损伤;肠绒毛内的 P 物质能神经纤维是肠粘膜屏障损伤的直接参与因素。     

烫伤大鼠肠道P物质含量和P物质能神经的观察分析

以３０％ＴＢＳＡⅢ度烫伤大鼠为模型，采用ＲＩＡ和ＩＨＣ的方法，动态观察了伤后７２小时大鼠空肠免疫活性Ｐ物质（ｉＳＰ）的含量及绒毛内Ｐ物质－免疫反应（ＳＰ－ＩＲ）阳性神经纤维的变化。结果发现：①烫伤大鼠空肠内ｉＳＰ伤后１小时出现显著升高，４小时仍处于高水平状态；８小时后明显下降，伤后７２小时一直处于低水平状态。②绒毛内ＳＰ－ＩＲ阳性神经纤维在形态、分布密度和纤维内ＳＰ－ＩＲ物质的含量都有明显的改变。图像分析定量结果表明：绒毛内ＳＰ－ＩＲ阳性神经纤维分布密度，伤后１小时明显增加，随后减少，４８小时再次出现增加；神经纤维内ＳＰ－ＩＲ阳性物质伤后１小时及４小时明显升高，１２小时明显下降，２４小时后再次升高。实验结果说明：烫伤后大鼠空肠内Ｐ物质出现应激性释放与消耗，Ｐ物质可能参与了烫伤后大鼠的肠道损伤；肠绒毛内的Ｐ物质能神经纤维是肠粘膜屏障损伤的直接参与因素。     

Comparative localization of heme oxygenase-2 and nitric oxide synthase in the autonomic innervation to the human ductus deferens and seminal vesicle

PURPOSE: The aim of present study was to determine the topographic relationship between heme oxygenase-2 (HO-2), which synthesizes carbon monoxide (CO), and neuronal nitric oxide synthase (nNOS), which generates nitric oxide (NO), in the autonomic nerves of the human ductus deferens and seminal vesicle. MATERIALS AND METHODS: Specimens of the ductus deferens and seminal vesicle were obtained during cancer surgery or vasectomy. HO-2 and nNOS were localized by indirect immunofluorescence. Additionally, the histochemical NADPH-diaphorase (NADPH-d) activity of NOS was demonstrated using a standard staining method and some modifications. RESULTS: Anti-HO-2 labeling stained virtually all nerve cell bodies in local ganglia of the pelvic plexus, which is composed of a mixed population of postganglionic sympathetic and parasympathetic neurons supplying the pelvic viscera. Furthermore, nerve cell bodies in the wall of the seminal vesicle, which are considered an extension of the pelvic plexus, were also found to stain positively for HO-2. Some of the HO-2-immunoreactive ganglion cells were also nNOS-positive, their proportion varying between individual ganglia but generally not exceeding 20%. Both enzymes were present in large adventitial nerve trunks. Only nNOS but no HO-2 was found in small intramuscular and mucosal nerve fibers. In both the ductus deferens and seminal vesicle, the highest density of nNOS-containing nerve fibers was in the lamina propria of the mucosa. A well-developed plexus of nNOS-positive nerve fibers was also observed in the muscular layer of the seminal vesicle. By contrast, there was a very sparse innervation by nNOS-positive nerve fibers in the muscle coat of the ductus deferens. In addition, a population of epithelial cells in the seminal vesicle may contain an isoform of NOS, as revealed by a resistant NADPH-d activity. CONCLUSIONS: These findings set the scene for functional studies which will hopefully clarify the biological role of CO and NO in the control of the ductus deferens and seminal vesicle.     

早期肠道喂养改善烧伤后肠道血液灌流的实验研究

目的 探讨早期肠道喂养改善烧伤大鼠肠血液灌流的机理。方法 采用３０％体表面积Ⅲ度烧伤大鼠模型,分为正常对照（Ｃ）、单纯烧伤（Ｂ）和早期喂养（ＶＩＰ）含量及肠粘膜血流量（ＬＭＢＦ）。结果 烧伤后肠组织中ＥＴ、ＮＯ、ＶＩＰ及ＥＴ／ＶＩＰ比值均呈上升趋势,而ＩＭＢＦ则显著低于伤前。ＥＦ组中ＥＴ含量及ＥＴ／ＮＯ、ＥＴ／ＶＩＰ比值明显低于Ｂ组,而ＮＯ、ＶＩＰ及ＩＭＢＦ则高于Ｂ组,肠道喂养烧伤后肠道缺血状况的