Ghrelin在大鼠神经系统中的表达对胃黏膜的保护作用

目的:研究大鼠脑、小肠肌间神经丛神经元和血浆内Ghrelin的表达,探讨Ghrelin在水浸加束缚应激性胃溃疡中的作用及机制．方法:选择健康♂Wiater大鼠76只,随机分为 6组:水浸加束缚组(10只);侧脑室注射Ghrelin 组(24只);皮下注射L-NAME 侧脑室注射 Ghrelin组(8只)和相应的3个对照组(正常大鼠组18只,侧脑室注射生理盐水组8只,皮下注射 L-NAME 侧脑室注射生理盐水组8只)．采用放射免疫分析、荧光免疫组化双染和神经生理学等实验方法,观察脑、小肠肌间神经丛和血浆内Ghrelin的表达,探讨Ghrelin对大鼠束缚加水浸诱导的应激性胃溃疡的影响及机制．结果:在正常大鼠小肠肌间神经丛内和原代培养的肠肌间神经丛神经元均可见有Ghrelin 样免疫反应阳性物(Ghrelin-IR)的表达,且 Ghrelin-IR与胆碱乙酰转移酶(CHAT)共同表达于同一神经元内,但Ghrelin-IR不与一氧化碳合酶(NOS)和消化道感觉性神经元内特有的钙结合蛋白(Calbindin,Calb)共存．在大鼠应激性胃溃疡产生的同时,其血浆内Ghrelin- IR的含量明显减少(198．3±29．6 ng/L vs 141．7 ±26．5 ng／L．P=0．026)．而下丘脑、延脑、垂体和肌间神经丛神经元内Ghrelin-IR的含量明显升高(分别为96．2±18．1 pg／mg vs 153．2 ±11．6 pg／mg．P=0．006;89．8±16．5 pg／mg vs 144．4±13．9 pg／mg,P=0．007;108．3± 11．9 pg／mg vs 198．2±23．3 pg／mg,P=0．002; 48．8±12．8 pg／mg vs 86．2±21．5 pg／mg．P= 0．02);侧脑室注射Ghrelin大鼠应激性胃溃疡的产生明显减少,且呈明显的量效依赖关系(溃疡指数:生理盐水,86．7±6．2;50 ng Ghrelin,79．3±10．7 P=2．18;500 ng Ghrelin, 61．3±11．7,P=0．04;5 000 ng Ghrelin,35．6 ±10．8,P=0．005),经sc一氧化氮合酶抑制剂 L-NAME后,Ghrelin的胃黏膜细胞保护作用消失．结论:Ghrelin与ChAT共存表达于肠肌间神经丛胆碱能神经元;应激性胃溃疡发生时,中枢神经系统和血浆内Ghrelin的表达发生变化; 中枢Ghrelin对胃黏膜细胞具有保护作用,且呈明显的量效依赖关系．     

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。     

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。     

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。     

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。     

慢性胰腺炎大鼠肠运动功能和肌间神经丛NOS阳性神经元的变化

目的观察大鼠慢性胰腺炎时肠运动功能和肌间神经丛NOS阳性神经元的变化,探讨慢性胰腺炎大鼠肠动力障碍的可能机制。方法 SD雄性大鼠20只随机分为假手术组(n=10)和慢性胰腺炎组(n=10),用质量浓度为30 g/L TNBS乙醇盐溶液逆行胰胆管灌注制作慢性胰腺炎模型;假手术组注射等体积生理盐水。造模成功后胃内灌服质量浓度为10 g/L台盼蓝溶液,测定小肠推进比。取胰腺组织进行HE染色,观察病理变化。取末段回肠制作肌间神经丛全层标本,应用双重免疫荧光染色法观察肌间神经丛神经元,计算NOS阳性神经元占总神经元的百分比。结果假手术组肌间神经丛NOS阳性神经元的比例为(29.40±2.03)%,慢性胰腺炎组肌间神经丛NOS阳性神经元的比例为(39.62±1.75)%,两组比较,差异有统计学意义(P0.05)。慢性胰腺炎组大鼠小肠推进比显著降低[(49.70±3.74)%vs(31.14±3.23)%,P0.05]。NOS阳性神经元比例与小肠推进比呈负相关(r=-0.853,P0.05)。结论慢性胰腺炎大鼠存在肠动力障碍,同时回肠肌间神经丛NOS阳性神经元发生了重塑,慢性胰腺炎肠动力障碍可能与NOS抑制性神经元的表达上调相关。     

肠易激综合征模型大鼠结肠肌间神经丛钙视网膜蛋白阳性神经元增加

目的检测肠易激综合征腹泻型(IBS-D)和便秘型(IBS-C)两种模型大鼠远端回肠、结肠黏膜下神经丛(SMP)和肌间神经丛(MP)钙视网膜蛋白(CALR)阳性神经元的变化,探讨其在IBS发病中的作用。方法分别采用慢急性联合应激、冰水灌胃法建立IBS-D和IBS-C大鼠模型,制作回肠和结肠SMP、MP全层铺片标本,免疫组织荧光双染法检测CALR阳性神经元的改变。结果与对照组相比,IBS-D和IBS-C模型大鼠回肠SMP、MP和结肠SMP中CALR阳性神经元比例的差异无统计学意义(P0.05),结肠MP中CALR阳性神经元的比例显著增加,分别为(17.2±3.2)%vs(13.4±2.9)%、(19.1±5.9)%vs(14.9±3.4)%,差异均有统计学意义(P0.05)。结论 IBS-D、IBS-C模型大鼠结肠MP中CALR阳性神经元增加可能参与IBS肠道敏感性增高。     

应激大鼠幽门区一氧化氮和神经元型一氧化氮合酶与胃内胆汁反流的关系

背景：幽门区一氧化氮（NO）与幽门括约肌舒张功能密切相关，但从形态学上探讨神经元型一氧化氮合酶（nNOS）与胃内胆汁反流关系的研究却较少。目的：探讨应激大鼠幽门区NO和nNOS含量变化对胃内胆汁反流的影响。方法：55只大鼠随机分为正常对照组（n=10）、实验组（n=30）和免疫组化组（n=15），后两组以水浸束缚应激诱导应激性溃疡模型。实验组于应激开始后2、4、5、6、8和10h各取5只大鼠，分别检测胃内胆汁酸浓度和胃黏膜溃疡指数（UI，Guth评分），以生化试剂盒检测幽门区NO含量。免疫组化组于应激开始后0、4和6h各取5只大鼠，以免疫组化SP法结合图像分析检测幽门区nNOS免疫反应阳性产物的平均光密度值。结果：应激性溃疡大鼠幽门区NO含量于应激结束后lh达到峰值（P〈0．01）；应激结束后2h，胃内胆汁酸浓度达到峰值（P〈0．01）；胃黏膜UI于应激结束后4h达到峰值（P〈0．01）。免疫组化染色显示应激性溃疡大鼠胃内胆汁反流增加时，幽门区肌间神经丛nNOS免疫反应阳性产物平均光密度值显著增高（P〈0．01）。结论：NO能促进应激大鼠胃内胆汁反流，NO和nNOS参与了幽门括约肌舒张功能的调控。     

腹泻型肠易激综合征模型大鼠结肠黏膜下神经丛神经元的改变

目的 检测慢急性联合应激腹泻型肠易激综合征(IBS-D)模型大鼠结肠黏膜下神经丛(SMP)神经元总数以及特异性标志物阳性神经元的变化,从肠神经系统(ENS)水平推断神经元改变在肠易激综合征(IBS)发病中的作用.方法 建立慢急性联合应激IBS-D大鼠模型,留取大鼠结肠制作SMP全层铺片标本.应用免疫组织荧光双染法检测SMP中神经元总数以及乙酰胆碱转移酶(CHAT)、血管活性肠肽(VIP)和一氧化氮合酶(NOS)阳性神经元数目和比例,评价IBS-D模型大鼠ENS神经元的改变.结果 IBS-D模型大鼠结肠SMP神经节和神经元总数无明显改变.与对照组相比,IBS-D模型大鼠结肠SMP中VIP阳性神经元比例明显增高(62.2%±6.2%比55.4%±5.4%,P<0.05),NOS阳性神经元比例亦明显增高(15.0%±4.0%比10.5%±2.9%,P<0.05),ChAT阳性神经元比例无明显改变.结论 IBS-D模型大鼠结肠SMP中VIP阳性神经元和NOS阳性神经元比例增高,提示在慢急性联合应激诱发1BS-D时,ENS中SMP神经元的变化可能通过增加肠道分泌而导致或加重腹泻症状.     

慢传输型便秘患者结肠中五羟色胺受体亚型的表达及意义

目的:研究主要5-HT受体亚型在慢传输型便秘(slowtransitconstipation,STC)患者结肠中的表达,探讨其在慢传输型便秘发病机制中的作用.方法:采用免疫组化EnVision法,检测20例STC患者和20例对照组结肠组织中5-HT1A,5-HT3和5-HT4受体的分布及表达水平,并采用IMS计算机辅助图像分析系统进行半定量分析.结果:5-H1A受体分布于黏膜下层、肌层,肌间神经丛5-HT1A受体的表达在STC组和对照组间无显著差异(P=0.548).5-HT3受体分布于黏膜下层和肌层,肌间神经丛STC组阳性指数显著低于对照组(65.2±15.9vs94.3±20.1,P<0.01).5-HT4受体分布于黏膜层、黏膜下层、肌层.在黏膜层和肌间神经丛,STC组5-HT4受体阳性指数均显著低于对照组(57.8±10.9vs78.5±12.9,P<0.01;77.5±19.9,119.2±26.9,P<0.01).STC组中,5-HT3受体表达水平与结肠传输试验第5天体内残留标志物数量无关(P>0.05);但5-HT4受体表达水平与第5天体内残留标志物数量呈负相关(r=-0.782,P<0.01).结论:STC患者结肠中存在5-HT3和5-HT4受体亚型的表达下调,两者可能参与了STC的发病机制.     

Altered colonic transit in TNBS-induced experimental colitis in guinea pig and distribution of nitric oxide synthase in the colonic wall]

BACKGROUND/AIMS: Inflammation-induced alterations in smooth muscle contractility may be due to the effects on smooth muscle itself, neurotransmitters or enteric nerves. In dextran sulfate sodium-induced colitic rat, the delay in colonic transit was caused by decreased activity and production of neuronal nitric oxide synthase (nNOS) in the myenteric plexus of the distal colon. The aim of this study was to investigate the relationship between the delay in colonic transit and the distribution of inducible NOS (iNOS) and nNOS immunoreactive cells in the myenteric plexus of trinitrobenzene sulfonic acid (TNBS)-induced colitic guinea pig. METHODS: Sacrificed and their colonic tissues of forty-five TNBS-induced colitic guinea pigs were used to measure the colonic transit, and analyzed by immunohistochemistry. RESULTS: Colonic transit was delayed significantly at 3, 7 and 14 days after administration of TNBS. In control, nNOS immunoreactivity was present in the mucosa, submucosa, lamina propria, and ganglion cells of the myenteric plexus, while after TNBS treatment, reduced nNOS cells were found. However, the number of nNOS ganglion cells in the myenteric plexus was similar to those seen in controls. After administration of TNBS, iNOS immunoreactivity was increased in the mucosa and submucosa, but the number of iNOS positive ganglion cells in the myenteric plexus was not changed compared to control. CONCLUSIONS: It is suggested that in TNBS-induced guinea pig colitis, delayed colonic transit is not associated with the expression of nNOS nor iNOS in the myenteric plexus.     

Oligoneuronal Hypoganglionosis in Patients with Idiopathic Slow-Transit Constipation

PURPOSE: Several alterations of the enteric nervous system have been described as an underlying neuropathologic correlate in patients with idiopathic slow-transit constipation. To obtain comprehensive data on the structural components of the intramural nerve plexus, the colonic enteric nervous system was investigated in patients with slow-transit constipation and compared with controls by means of a quantitative morphometric analysis. METHODS: Resected specimens were obtained from ten patients with slow-transit constipation and ten controls (nonobstructive neoplasias) and processed for immunohistochemistry with the neuronal marker Protein Gene Product 9.5. The morphometric analysis was performed separately for the myenteric plexus and submucous plexus compartments and included the quantification of ganglia, neurons, glial cells, and nerve fibers. RESULTS: In patients with slow-transit constipation, the total ganglionic area and neuronal number per intestinal length as well as the mean neuron count per ganglion were significantly decreased within the myenteric plexus and external submucous plexus. The ratio of glial cells to neurons was significantly increased in myenteric ganglia but not in submucous ganglia. On statistical analysis, the histopathologic criteria (submucous giant ganglia and hypertrophic nerve fibers) of intestinal neuronal dysplasia previously described in patients with slow-transit constipation were not completely fulfilled. CONCLUSION: The colonic motor dysfunction in slow-transit constipation is associated with quantitative alterations of the enteric nervous system. The underlying defect is characterized morphologically by oligoneuronal hypoganglionosis. Because the neuropathologic alterations primarily affect the myenteric plexus and external submucous plexus, superficial submucous biopsies are not suitable to detect these innervational disorders.     

Pathophysiological significance of neuronal nitric oxide synthase in the gastrointestinal tract

It has been demonstrated that nitric oxide (NO) is a major inhibitory nonadrenergic, noncholinergic (NANC) neurotransmitter in the gastrointestinal (GI) tract. NO released in response to nerve stimulation of the myenteric plexus causes relaxation of the smooth muscle. NO is synthesized by the activation of neuronal NO synthase (nNOS) in the myenteric plexus. Released NO plays an important physiological role in various parts of the GI tract. NO regulates the muscle tone of the sphincter in the lower esophagus, pylorus, sphincter of Oddi, and anus. NO also regulates the accommodation reflex of the fundus and the peristaltic reflex of the intestine. Previous studies have shown that NOS inhibitors delay gastric emptying and colonic transit. The reduction of nNOS expression, associated with impaired local production of NO, may be responsible for motility disorders in the GI tract. There is accumulated evidence that dysfunction of NO neurons in the myenteric plexus may cause various GI diseases. These reports are reviewed and possible mechanisms of altered nNOS expression are discussed in this article. In particular, impaired nNOS synthesis of the myenteric plexus seems to be an important contributing factor to the pathogenesis of achalasia, diabetic gastroparesis, infantile hypertrophic pyloric stenosis, Hirschsprung's disease, and Chagas' disease. Reduced NO release and/or nNOS expression are suspicious in a subset of patients with functional dyspepsia. Although the etiology of intestinal pseudo-obstruction remains unknown, it is conceivable that extrinsic denervation may upregulate nNOS expression, resulting in enhanced muscular relaxation and disturbed peristalsis. An animal model of colitis showed impaired nNOS expression in the colonic myenteric plexus. Antecedent infection may be associated with the impaired NO pathways observed in functional dyspepsia, colitis, and Chagas' disease.     

Impaired nitrergic innervation in rat colitis induced by dextran sulfate sodium

BACKGROUND & AIMS: The pathophysiological role of neuronal nitric oxide synthase (nNOS) in colitis remains unknown. METHODS: We investigated colonic transit, nonadrenergic, noncholinergic (NANC) relaxation, nNOS activity, and nNOS synthesis in the myenteric plexus in dextran sulfate sodium (DSS)-induced colitis in rats. RESULTS: Oral administration of 5% DSS for 7 days induced predominant distal colitis and delayed colonic transit. In the proximal colon, carbachol-, sodium nitroprusside-, and electrical field stimulation (EFS)-induced responses were not different between control and DSS-treated rats. In the distal colon, EFS-evoked cholinergic contraction, NANC relaxation, and orphanin FQ-induced contraction were significantly impaired in DSS-treated rats compared with those in control rats, but carbachol- and sodium nitroprusside-induced responses remained intact in DSS-treated rats. The number of nNOS-immunopositive cells, catalytic activity of NOS, and nNOS synthesis in the colonic wall were significantly reduced in the distal colon of DSS-treated rats. In contrast, the number of PGP 9.5-immunopositive cells and PGP 9.5 synthesis in the colonic wall remained intact in the distal colon of DSS-treated rats. CONCLUSIONS: These results suggest that impaired NANC relaxation in the distal colon is associated with reduced activity and synthesis of nNOS in the myenteric plexus in DSS-induced colitis.     

Expression of the P2X2 receptor in different classes of ileum myenteric neurons in the female obese ob/ob mouse

AIM:To examine whether the ob/ob mouse model of obesity is accompanied by enteric nervous system ab-normalities such as altered motility METHODS:The study examined the distribution of the P2X 2 receptor (P2X 2 R) in myenteric neurons of female ob/ob mice. Specifically, we used immunohistochemistry to analyze the co-expression of the P2X 2 R with neuronal nitric oxide synthase (nNOS), choline acetyltrans-ferase (ChAT), and calretinin (CalR) in neurons of the small intestine myenteric plexus in ob/ob and control female mice In these sections, we used scanning confocal microscopy to analyze the co-localization of these markers as well as the neuronal density (cm 2 ) and area profile (μm2) of P2X 2 R-positive neurons In addition, enteric neurons were labeled using the nicotinamide adenine dinucleotide (NA H) diaphorase method and analyzed with light microscopy as an alternate means by which to analyze neuronal density and areaRESULTS:In the present study, we observed a 29 6% increase in the body weight of the ob/ob animals (OG) compared to the control group (CG) In addition, the average small intestine area was increased by approxi-mately 29 6% in the OG compared to the CG Immu-noreactivity (IR) for the P2X 2 R, nNOS, ChAT and CalR was detectable in the myenteric plexus, as well as in the smooth muscle, in both groups This IR appeared to be mainly cytoplasmic and was also associated with the cell membrane of the myenteric plexus neurons, where it outlined the neuronal cell bodies and their processes P2X 2 R-IR was observed to co-localize 100% with that for nNOS, ChAT and CalR in neurons of both groups In the ob/ob group, however, we observed that the neuronal density (neuron/cm 2 ) of P2X 2 R-IR cells was in-creased by 62% compared to CG, while that of NOS-IR and ChAT-IR neurons was reduced by 49% and 57%, respectively, compared to control mice The neuronal density of CalR-IR neurons was not different between the groups Morphometric studies further demonstrated that the cell body profile area (μm2) of nNOS-IR, ChAT-IR and CalR-IR neurons was increased by 34%, 20% and 55%, respectively, in the OG compared to controls Staining for NA H diaphorase activity is widely used to detect alterations in the enteric nervous system; however, our qualitative examination of NA H-diaphorase positive neurons in the myenteric ganglia revealed an overall similarity between the two groups CONCLUSION:We demonstrate increases in P2X2R expression and alterations in nNOS, ChAT and CalR IR in ileal myenteric neurons of female ob/ob mice compared to wild-type controls.     

Ucn3及其受体CRFR2在肠易激综合征大鼠模型肠神经系统中的表达

目的:探讨尿皮质素3(Urocotin3,Ucn3)及其受体促肾上腺皮质释放因子受体2(corticotrophin releasing factor receptor2,CRFR2)在肠易激综合征中的表达.方法:将36只180-220g的Wistar大鼠随机分为对照组(N)、急性应激组(A,急性束缚1h)、慢性应激组(C,28d不可预知轻度应激)、急慢性联合应激组(AC,在慢性应激基础上给予急性束缚)4组建模.采用排便粒数、敞箱行为评分和蔗糖水偏嗜度评价动物模型.建成后留取大鼠结肠组织,采用Real-timePCR方法检测各组大鼠结肠中Ucn3及其受体CRFR2表达水平的变化.结果:Ucn3在各组大鼠结肠中的表达:N组1.108±0.293,A组3.594±1.839,C组1.852±0.674,AC组3.989±1.591,各应激组Ucn3的表达均高于对照组(P<0.05),各应激组间A组vsC组(P<0.017),C组vsAC组(P<0.002),表达有统计学差异.CRFR2在各组大鼠结肠中的表达:N组1.042±0.217,A组2.119±0.468,C组1.568±0.507,AC组2.392±0.840,各应激组CRFR2的表达均高于对照组(P<0.05).各应激组之间没有统计学差异.结论:慢性应激、慢急性联合应激建立肠易激综合征大鼠模型重复性好.Ucn3及其受体CRFR2在肠易激综合征中表达升高,且Ucn3在急性应激后升高比慢性应激后更明显.     

IL-4、IL-13在慢性综合应激模型大鼠Cajal细胞损伤中的作用

目的观察慢性综合应激模型大鼠中IL-4及IL-13的表达变化,并初步探讨其在Cajal细胞(Interstitial cells of Cajal,ICC)损伤中的作用。方法健康成年SD雄性大鼠20只,随机分为实验组和对照组,每组10只,实验组大鼠制备慢性综合应激模型,旷场实验鉴定造模成功后,采用ELISA法测定血清IL-4、IL-13的浓度,免疫荧光法观察肠道肌间神经丛TMEM16A的表达与分布。结果模型组和对照组大鼠血清IL-4浓度分别为(8.09±0.97)pg/mL和(6.98±0.69)pg/mL,差异有统计学意义(t=3.363,P<0.01),血清IL-13浓度分别为(5.96±0.67)pg/mL和(5.26±0.73)pg/mL,差异有统计学意义(t=2.322,P<0.05);荧光显微镜下观察肠道平滑肌间神经丛TMEM16A有阳性表达,模型组在不同节段肠道肌间神经丛为阳性表达的细胞减少且分布稀疏。结论慢性综合应激时,IL-4、IL-13等Th2免疫应答相关炎性因子产生增加,其可能通过调控TMEM16A表达参与了ICC的损伤。     

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.