温胃阳汤对胃动力低下大鼠胃电活动、胃窦肌间神经丛5-羟色胺能神经及其受体的影响

[目的]观察温胃阳汤对胃动力低下大鼠胃电活动、胃窦肌间神经丛5-羟色胺（5-HT）能神经及其受体的影响.[方法]将70只Wistar大鼠随机分为正常组（10只,雌雄各半）和模型组（60只,雌雄各半）,模型组每只大鼠按10 g/kg甘草煎剂灌胃,随机分为模型3d组、7d组、温胃阳汤低、中、高剂量组及吗丁啉组.温胃阳汤低、中、高各剂量组分别予浓度为0.743 g/ml、1.485 g/ml、2.970 g/ml的温胃阳汤灌胃,吗丁啉组予0.27 mg/ml吗丁啉混悬液（按100 g体重1 ml）灌胃.观察各组大鼠胃电活动的变化,并以免疫组化方法结合图像分析技术分析5-HT免疫反应阳性产物的分布、数量和免疫反应强度.以免疫印迹法分析大鼠胃窦组织中5-羟色胺受体（5-HTR）,计算出各组的蛋白相对表达量,以百分数表示各组的蛋白表达量.[结果]一定剂量的甘草煎剂能明显抑制大鼠胃电活动,使大鼠窦组织5-HT免疫反应阳性神经纤维、神经节内阳性神经细胞体和平均光密度值明显减少,5-HTR的表达显著降低.而温胃阳汤低、中、高剂量组及吗丁啉组均可使胃动力低下大鼠胃窦组织5-HT免疫反应阳性神经纤维、神经节内阳性神经细胞体和带膨体的神经纤维增多,5-HTR的表达亦得到提高;其中中、高剂量组效果与吗丁啉组比较差异无统计学意义,与正常组比较差异有统计学意义（P＜0.05或P＜0.01）.[结论]经温胃阳汤治疗后,胃窦肌间神经丛5-HT免疫反应阳性神经纤维和神经细胞体分布增多,5-HTR表达升高,5-HT能神经活性增强,表明温胃阳汤能通过5-HT能神经参与促胃动力作用.     

肝衰竭大鼠胃动力的改变及其机制的初步探讨

[目的]观察肝衰竭大鼠胃动力的变化并探讨其初步机制。[方法]40只Wistar大鼠随机分为肝衰竭模型组和对照组,采用葡聚糖蓝-2000为标记物观察大鼠胃排空的变化,应用乙酰胆碱酯酶(AchE)和NADPH-d组织化学染色(NOS)及肌间神经丛全层铺片技术,观察肝衰竭大鼠胃窦肌间神经丛胆碱能和氮能神经的变化,免疫组化染色观察胃窦C-kit阳性Cajal间质细胞的变化,并进行定量分析。[结果]与对照组比较,模型组大鼠胃排空明显减弱,胃窦肌间神经丛胆碱能阳性神经元数量减少、神经纤维变细、分布较稀疏,2组比较差异有统计学意义(P<0.01);模型组氮能神经阳性神经元数量及神经纤维分布明显高于对照组(P<0.01);胃窦C-kit阳性Cajal间质细胞明显少于对照组(P<0.01)。[结论]肝衰竭大鼠胃动力明显减退,其机制与胃窦肌间神经丛胆碱能神经分布减少、Cajal间质细胞减少及氮能神经分布增加有关。     

胃起搏对大鼠胃窦肌间神经丛单胺氧化酶mRNA表达的影响

目的研究胃窦肌间神经丛中单胺氧化酶mRNA表达,探讨5羟色胺(5-HT)在胃起搏机制中的作用。方法通过手术建立Wistar大鼠胃起搏模型(近远端胃各缝制一对电极),分为起搏组(n=10)和对照组(n=6)。采用逆转录聚合酶链反应(RT-PCR)检测起搏组和对照组胃窦肌间神经丛中MAO-A mRNA、MAO-B mRNA的表达量,以恒定表达的β-actin作为内参照。计算MAO-AmRNA、MAO-B mRNA与β-actin mRNA表达积分光密度值的比例(MA/B,MB/B),以反映组织中MAO-AmRNA、MAO-B mRNA的相对表达量。结果RT-PCR研究显示起搏组MAO-A mRNA表达显著弱于对照组,起搏组MA/B比值明显较对照组较少(0.37±0.11vs0.95±0.57,P<0.001);而起搏组MAO-B mRNA表达与对照组无差别,两组间MB/B比值比较也无显著性意义(0.97±0.24vs1.01±0.58,P>0.05)。结论胃起博后胃肌间神经丛内单胺氧化酶mRNA表达量明显减少,表明胃窦肌间神经丛内5-HT可能在胃起搏中发生了重要作用。     

肝衰竭大鼠胃窦胆碱能和氮能神经分布的变化分析

目的观察肝衰竭大鼠胃排空及胃窦肌间神经丛胆碱能和氮能神经的变化。方法 40只Wistar大鼠随机分为肝衰竭模型组和对照组,采用葡聚糖蓝-2000为标志物观察大鼠胃排空的变化,应用乙酰胆碱酯酶(AchE)和还原型辅酶Ⅱ硫辛酰胺脱氢酶(NADPH-d)组织化学染色及肌间神经丛全层铺片技术,观察肝衰竭大鼠胃窦肌间神经丛胆碱能和氮能神经的变化,并进行定量分析。计量资料以均数±标准差(x±s)表示,组间比较采用t检验。结果肝衰竭组大鼠胃排空明显减弱(163.00±25.68 vs 100.00±18.93,P0.01),胃窦肌间神经丛胆碱能阳性神经元数量减少,神经纤维变细,分布较稀疏,明显低于对照组(t=3.201,P0.01);氮能神经阳性神经元数量及神经纤维分布明显高于对照组(t=2.912,P0.01)。结论肝衰竭大鼠胃功能的减退与胃窦肌间神经丛胆碱能神经分布减少及氮能神经分布增加有关。     

胃电刺激对大鼠十二指肠部分肠神经递质释放的影响

背景：胃电刺激（GES）对胃动力的影响已引起广泛关注，但关于GES后十二指肠肌间丛神经及其递质的变化。目前所知尚少。目的：研究GES对大鼠十二指肠壁内乙酰胆碱（Ach）、一氧化氮（NO）、P物质（SP）和血管活性肠肽（VIP）释放的影响。方法：建立Wistar大鼠GES模型，将模型大鼠分为GES组（n=10）和对照组（n=6）。选用适宦的刺激参数控制GES组胃慢波，应用免疫组化方法结合图像分析技术分析十二指肠肌间神经丛胆碱能、NO能、SP能和VIP能神经活性的变化。结果：GES组十二指肠肌间神经丛胆碱乙酰转移酶（ChAT）、神经元划一氧化氮合酶（nNOS）免疫反应阳性纤维较对照组明显增多、染色增强。易见ChAT、nNOS免疫反应阳性神经节和阳性神经元细胞体；而SP、VIP免疫反应阳性纤维和末梢及其染色强度在GES后无明显变化。GES组肌删种经从ChAT、nNOS免疫反应阳性产物的平均光密度值显著高于对照组（P〈0．001），SP、VIP免疫反应阳性产物的平均光密度值则与对照绀无显著差异（P〉0．05）。结论：GES后大鼠十二指肠壁内Ach、NO释放增多，VIP、SP则无明显变化。     

糖尿病大鼠胃窦胆碱能神经元与胃动力障碍的关系

目的 探讨链脲佐菌素(STZ)-糖尿病大鼠胃动力障碍和胃肌间神经丛胆碱能之间的关系.方法 45只SD大鼠随机分为对照组、糖尿病组和胰岛素组.成模后16 w测定大鼠胃动力,观察胃肌间神经丛胆碱能神经元的形态变化.结果 与对照组比较,糖尿病组大鼠胃动力减弱(P<0.01),胃窦肌间神经丛胆碱能神经元计数显著降低(P<0.01).与糖尿病组相比较,胰岛素组胃动力显著增高 (P<0.05),胃窦肌间神经丛胆碱能神经元平均光密度显著增高(P<0.05),胆碱能神经元计数有改善的趋势(P>0.05).结论 STZ-糖尿病大鼠胃动力障碍可能与胃肌间神经丛胆碱能神经损伤有关,胰岛素治疗能在一定程度上改善糖尿病胃动力障碍.     

胃电节律失常大鼠胃窦肌间神经丛胆碱能神经的改变

目的：探讨大鼠胃窦肌间神经丛胆碱能神经，氮能神经含量变化与胃电节律失常的关系。方法：63例大鼠随机分为正常对照组，胃电节律失常模型组和白芍组。饲养4周后记录并分析胃电信号，测定胃窦肌间神经丛胆碱能神经含量。结果：模型组胃电节律失常明显增加，胃窦肌间神经丛胆碱能神经含量减少；经白芍治疗后，胃电节律失常明显减少，胃窦肌间神经丛胆碱能神经含量恢复正常。结论：胃窦肌间神经丛的胆碱能神经与胃电节律关系密切，当胆碱能神经减少时，胃电节律失常明显增加。     

交感神经和肌间神经丛在胃电刺激调控胃慢波中的作用

目的研究胃交感神经及肌间神经丛在电刺激调控胃慢波活动中的作用,确定胃电起博的神经机制和作用环节,为今后起搏器的深入研究打下基础。方法10只雄性wistar大鼠随机分为对照组和电刺激组,各5只。全部大鼠植入浆膜电极,电刺激组大鼠行胃电刺激至胃慢波被完全控制。植入电极组不行电刺激。采用免疫组化S P法检测并比较两组大鼠胃窦肌间神经丛和脊髓后角C fos蛋白表达。结果电刺激组大鼠胃慢波全部被完全控制,所需能量为2 70±80 .6ms ,2mA。2组大鼠脊髓中间内侧核,中间外侧核均未见C fos阳性神经元,而后角浅层均见散在C fos表达,比较无显著性差异(P >0 .0 5 )。植入电极组胃窦肌间神经丛未见C fos阳性神经元,电刺激组胃窦肌间神经丛可见C fos阳性神经元。结论适宜参数的胃电刺激可完全控制大鼠胃慢波。肌间神经丛参与胃电刺激调控胃慢波,而交感神经则无明显作用。     

突触在胃起搏调控胃慢波活动中的作用

目的研究胃窦肌间神经丛中突触素（synaptophysin,Syn）分布及其mRNA和蛋白的表达,探讨突触素在胃起搏机制中的作用。方法通过手术建立Wistar大鼠胃起搏模型（近远端胃各缝制一对电极）,分为起搏组（GES组,n=10）和对照组（n=6）。应用免疫组化染色观察GES组和对照组胃窦间神经丛中突触素分布,采用逆转录聚合酶链反应（RT-PCR）和Western blot分别检测GES和对照组胃窦肌间神经丛中Syn mRNA和蛋白的表达量,以恒定表达的β-actin作为内参照。分别计算Syn mRNA与β-actinmRNA、Syn蛋白与β-actin蛋白表达积分光密度值的比值（Syn/β-actin）,以反映组织中Syn mRNA和蛋白的相对表达量。结果与对照组相比,GES组肌间神经丛中Syn免疫反应阳性产物分布显著增多（806.421±342.135vs448.3±261.467,P〈0.05;0.019±0.008vs0.010±0.005,P〈0.05）。RT-PCR研究显示GES组Syn mRNA表达显著强于对照组,GES组syn/β-actin比值明显较对照组增加（0.146±0.0 28vs0.082±0.025,P〈0.05）;Western blot研究显示GES组Syn蛋白表达也较对照组明显增强,两组Syn/β-actin比值比较差异有统计学意义（0.502±0.098vs0.298±0.018,P〈0.05）。结论胃起搏后胃肌间神经丛内突触素分布增多,突触素mRNA和蛋白表达量明显增加,表明胃窦肌间神经丛内突触可能在胃起搏中发挥了重要作用。     

糖尿病大鼠小肠肌间神经丛氮能神经的变化及其意义

目的探讨氮能神经在糖尿病大鼠小肠运动障碍中的作用。方法将50只大鼠随机分为正常对照组和槠尿病模型组,用链尿佐菌素建立大鼠糖尿病模型。3月后测定小肠传输速度,行小肠肌间神经丛氮能神经计数。结果糖尿病模型组大鼠小肠传输速度明显延迟,小肠肌间神经丛氮能神经细胞数和氮能神经神经节均明显减少。结论糖尿病小肠肌间神经丛氮能神经改变足导致小肠传输速率减慢的原因之一,从而引起小肠运动障碍。     

Origins of peptide and norepinephrine nerves in the mucosa of the guinea pig small intestine

Norepinephrine, acetylcholine, and certain peptides are contained in mucosal nerves and have potent effects on transepithelial water and electrolyte fluxes. It is difficult to ascribe roles for these nerves as their sources are unknown. The present studies were undertaken to determine the origins of nerve fibers that are found in the mucosa of the guinea pig small intestine and which contain one of the following substances: vasoactive intestinal peptide, substance P, somatostatin, neuropeptide Y, cholecystokinin, or norepinephrine. Nerve fiber origins were ascertained by making lesions to sever pathways through which the nerves could reach the mucosa. The lesioning operations were homotopic autotransplants of short (2 cm) segments of intestine; myectomies, in which a 5-10-mm length of intestine was stripped of longitudinal muscle and myenteric plexus; and extrinsic denervation, in which nerves reaching the intestine through the mesentery were severed. The results of these studies, considered along with previously published work, led to the upcoming conclusions. Nerve fibers in the mucosa showing immunoreactivity for vasoactive intestinal peptide, somatostatin, cholecystokinin, and neuropeptide Y arise from cell bodies in the overlying submucous plexus. Substance P fibers arise in part from the overlying submucous plexus and in part from the overlying myenteric plexus. Mucosal norepinephrine fibers arise from extrinsic sympathetic ganglia. Enkephalin, gastrin-releasing peptide, and 5-hydroxytryptamine, which are in some enteric nerves, are not found in submucous nerve cells and few, if any, fibers containing these substances supply the mucosa. Thus, the mucosa receives a dense nerve supply, much of which arises locally from submucous ganglia.     

Expression of c-fos in gastric myenteric plexus and spinal cord of rats with cervical spondylosis

AIM: To determine the expression of c-fos in gastric myenteric plexus and spinal cord of rats with cervical spondylosis and its clinical significance. METHODS: A cervical spondylosis model was established in rats by destroying the stability of cervical posterior column, and the cord segments C4-6 and gastric antrum were collected 3, 4 and 5 mo after the operation. Rats with sham operation were used as controls, c-fos neuronal counter-staining was performed with an immunohistochemistry method. Every third sections from C4-6 segments were drawn. The 10 most labeled c-fos-immunoreactive (Fos-IR) neurons were counted, and the average number was used for statistical analysis. The mean of Fos-IR neurons in myenteric plexus was calculated after counting Fos-IR neurons in 25 ganglia from each antral preparation, and expressed as a mean count per myenteric ganglion. RESULTS: There were a few c-fos-positive neurons in the cervical cord and antrum in the control group. There was an increased c-fos expression in model group 3, 4 and 5 mo after operation, whereas there was no significant increase in c-fos expression in the control group at 3, 4 and 5 mo. More importantly, there was a significant difference in c-fos expression between rats followed up for 3 mo and those for 5 mo in the model group (11.20±2.26 vs 27.68±4.36, P<0.05, for the cervical cord; and 11.3±2,3 vs 29.3±4.6, P<0.05, for the gastric antrum). There was no significant difference between rats followed up for 3 mo and those for 4 mo and between rats followed up for 4 mo and those for 5 mo in the model group. CONCLUSION: c-fos expression in gastric myenteric plexus was dramatically associated with that in the spinal cord in rats with cervical spondylosis, suggesting that the gastrointestinal function may be affected by cervical spondylosis. If this hypothesis is confirmed by further studies, functional gastrointestinal diseases such as functional dyspepsia and irritable bowel syndrome could be explained by neurogastroenterology.     

Serotoninergic neurons in human fetal intestine: an immunohistochemical study

The distribution of 5-hydroxytryptamine-like immunoreactivity was studied in whole-mount preparations of intestine from human fetuses. Immunoreactive nerve cell bodies were located in the myenteric plexus and were occasionally found in the submucous plexus; they were often seen to have long processes. Varicose fibers were found in the ganglia and internodal strands of the myenteric and submucous plexuses, in the deep muscular plexus of the circular muscle, and in the walls of some small mesenteric blood vessels immediately outside the intestine. This study provides evidence for the presence of serotoninergic nerves in the human intestine.     

胃电刺激对大鼠胃窦肌间神经丛Cajal间质细胞超微结构的影响

背景：前期研究发现胃窦肌间神经丛Caial间质细胞（ICC-MY）数量增多参与了长时程长脉冲胃电刺激（GES）对胃慢波的调控。目的：观察不同时程长脉冲GES对大鼠胃窦ICC-MY数量和超微结构的影响,进一步探讨GES调控胃慢波的可能机制。方法：建立Wistar大鼠GES模型。将模型大鼠分为3组,GES1组和GES2组选用适宜的刺激参数控制胃慢波,对照组不予GES。GES1组仅予刺激一次;GES2组每天刺激一次,连续20 d。完成GES后处死大鼠,取胃窦组织,行透射电子显微镜观察。结果：GES1组胃窦ICC-MY数量与对照组相比未见明显变化,GES2组ICC-MY数量较GES1组和对照组明显增多。GES1组和GES2组ICC胞质内线粒体和核糖体均较对照组增多。GES1组ICC突起与周围平滑肌细胞（SMC）直接相连,GES2组ICC与周围SMC紧密相连,对照组ICC与周围SMC之间未见明显连接。结论：胃窦ICC-MY数量和超微结构改变参与了长时程长脉冲GES对胃慢波的调控。     

Selective damage of intrinsic calcitonin gene-related peptide-like immunoreactive enteric nerve fibers in streptozotocin-induced diabetic rats

The distribution of calcitonin gene-related peptidelike immunoreactive (CGRP-LI) nerve fibers in the myenteric plexus of ileum and proximal colon of rats 8 wk after induction of diabetes with streptozotocin was studied using immunohistochemical techniques. A marked decrease in CGRP-LI nerve fibers mainly around the ganglion cells of the myenteric plexus of both ileum and proximal colon was observed in diabetic rats. The sparsely located immunoreactive nerve cell bodies in the control rats were absent in the diabetic preparations. There were, however, intensely stained CGRP-LI varicose nerve fibers that ran through the internodal strands and over the myenteric ganglia of the diabetic intestines. These findings indicate the presence of CGRP-LI nerve fibers of dual origin in the intestinal wall. The absence of positive cell bodies and diminished CGRP-LI nerve fibers around the ganglion cells in the diabetic tissues suggest that the state of diabetes selectively affects CGRP-LI nerve fibers of intrinsic rather than extrinsic origin. Furthermore, the absence of change in substance P-like immunoreactivity in the enteric system of rats with streptozotocin-induced diabetes of the same duration suggests that calcitonin gene-related peptide and substance P are contained in different populations of intrinsic nerve fibers in the gastrointestinal tract of the rat.     

胆囊收缩素在胃电节律失常中的作用及其神经病理学机制

目的：探讨胆囊收缩素（CCK）在胃电节律失常中的作用及其神经学机制。方法：在建立胃窦肌间神经丛铺片方法的基础上,用酶组织化学与免疫细胞化学方法,观察胃电节律失常大鼠胃窦肌间神经丛内胆碱能（Ach）神经、一氧化氮合酶（NOS）神经及CCK神经的变化。结果：模型组和CCK组大鼠均出现胃电节律失常,异常节律指数及慢波频率变异系数均显著高于正常组（P＜0．01）;模型组和CCK组NOS神经显著增加,Ach神经含量显著减少（P＜0．01）。结论：外源性及内源性CCK增加,能诱发胃电节律失常。CCK通过激活NOS,产生胃电节律失常。胃窦肌间神经丛神经中CCK及NOS神经含量异常增加,Ach神经减少是发生胃电节律失常的神经病理学机制之一。     

大鼠泻剂结肠肠壁神经生长因子受体p75的表达及其意义

背景：神经生长因子（NGF）及其受体与肠神经系统关系密切，演剂结肠有肠壁神经丛损害，但NGF受体p75在泻剂结肠中的表达和作用尚不明确。目的：研究NGF受体p75在正常大鼠和泻剂结肠大鼠中的表达及其在泻剂结肠形成中的意义。方法：采用大黄和酚酞建立泻剂结肠大鼠模型，以墨汁推进试验测定其传输功能：采用免疫组化法对正常大鼠和泻剂结肠大鼠的结肠肠壁进行p75检测。观察其在肠壁中的分布和表达情况。结果：与对照组相比，模型组肠道传输功能明显减慢，大黄组和酚酞组黑染肠管长度和百分比（黑染肠管长度／肠管总长度）均较对照组显著减低（P〈0．01，P〈0．05）。p75存正常大鼠结肠黏膜下神经丛中呈阳性表达，在肌间神经丛中多呈弱阳性表达。大黄组中p75表达明显增强，黏膜下神经丛亦呈强阳性表达，与对照组相比有显著差异（P〈0．01）；肌间神经从中多呈阳性表达（P〈0．05）。酚酞组黏膜下神经丛呈阳性表达，肌间神绎丛3只呈阳性表达，余表现为弱阳性或阴性，与对照组相比无明显差异。结论：p75在泻剂结肠中的异常表达可能参与肠神经丛冲经元细胞的退化变性或凋亡．从而引起泻剂结肠的肠神经系统病理变化，进一步导致结肠动力异常。这种损害与长期应用刺激性泻剂有关。