大鼠睾丸肥大细胞的发育和增龄性变化

用组织化学和形态计量学等方法研究正常大鼠生后到老年睾丸肥大细胞的类型及数量的变化。结果表明,正常大鼠睾丸肥大细胞３０天开始出现,位于白膜或白膜下,随年龄增加其数量逐渐增多,老年鼠（１８～２４月）增多较为明显,其面数密度、数密度及总数与幼年（３０～４５天）或成年（３～６月）鼠比较均有高度显著性（Ｐ＜０．０１）。睾丸肥大细胞（直径约１０μｍ）较结缔组织肥大细胞（直径１５～１８μｍ）小,异染颗粒少,Ａｌｃｉａｎｂｌｕｅ／ＳａｆｒａｎｉｎＯ染色时呈蓝染型或蓝染为主的间染型,硫酸小檗碱染色呈中等强度的黄色荧光。     

不同动物肥大细胞光镜的结构观察

研究不同种间肥大细胞的异同,为减轻排异反应提供了一定的理论依据。实验用甲醛固定,甲苯胺蓝染色。实验结果显示不同种的肥大细胞形态学结构不尽相同。禽类的肥大细胞与哺乳动物的相似,呈圆形、卵圆形,胞浆内含丰富的异染颗粒是其最显著的形态学特征。牛羊等的肥大细胞在低倍镜下形态多样,呈长梭形、椭圆形、不规则形,有的细胞表面可见细胞突起。     

牛蛙肥大细胞的组织化学与形态学

目的鉴定牛蛙组织中肥大细胞的存在。方法用于肥大细胞研究的一些常规组织化学技术与形态学方法。结果牛蛙的舌、肠、肠系膜和脾中肥大细胞数量较多,少量也见于神经、心、肾、肝和皮肤等多种组织中。肥大细胞有沿血管周和神经分布的倾向。脾脏中的肥大细胞形状比较一致,呈圆形或卵圆形,而在其它部位的肥大细胞则形态多样。Bouin氏液及Carnoy氏液是牛蛙肥大细胞优良的固定液。然而,与哺乳动物的黏膜肥大细胞相似的是,中性缓冲福尔马林(NBF)固定显著的阻断了牛蛙肠黏膜肥大细胞(MMC)的染色。有趣的是,甲苯胺蓝是牛蛙肥大细胞的最佳染料,它比阿尔新蓝能很好地显示牛蛙的肥大细胞。透射电镜下证实,牛蛙肥大细胞中含有大量特征性的胞浆颗粒。肥大细胞靠近雪旺氏细胞,并可见于神经束膜间,甚至以其突起与神经束膜相连。结论通过组织化学与形态学研究证实了牛蛙组织中肥大细胞的存在,再次证实肥大细胞与外周神经之间存在密切的解剖学关系。     

肥大细胞活化及生存

肥大细胞受各种刺激物的激活,脱颗粒释放生物活性介质,并藉此参与多种人类疾病。与其他炎症细胞不同,活化后的肥大细胞可继续生存。目前已知某些因素与肥大细胞的长寿相关,如细胞因子、某些分泌型磷脂酶A2(sPLA2)、IgE单体(mIgE)及Bcl-2家族基因A1。深入研究肥大细胞活化后生存的机制将有助于提供治疗肥大细胞相关疾病的有效靶干预。本综述了肥大细胞的活化及生存的研究动态。     

致炎剂联合降钙素基因相关肽对大鼠硬脑膜神经源性炎症的影响

目的：探讨致炎剂联合降钙素基因相关肽对大鼠硬脑膜神经源性炎症的影响,寻求最佳慢性偏头痛的实验模型。方法：24只SD雄性大鼠。随机分为生理盐水组（NS）、降钙素基因相关肽组1（CGRe1,10-3M）、降钙素基因相关肽组2（CGRP2,10-4M）,以及IS（20μL）＋CGRP（10μL,10-4M）组,每组各6只,通过甲苯胺蓝染色法,观察肥大细胞脱颗粒,采用伊文氏蓝荧光显像法,观察大鼠硬脑膜血管渗出,采用多普勒激光血流仪检测法,观察各组硬脑膜动脉血流量变化。结果：与NS组相比,CGRPl、CGRP2、IS＋CGRP组肥大细胞脱颗粒数和比率、脑血流量均明显升高,P〈0．05,并且硬脑膜荧光红斑明显增多;与CGRPl、CGRP2组相比,IS＋CGRP组肥大细胞脱颗粒数和比率、脑血流量均明显升高,P〈0．05,并且硬脑膜荧光红斑明显增多。CGRP两组相比,上述指标比较,差异没有统计学意义,P〉0．05。结论：IS＋CGRP能够明显刺激大鼠硬脑膜发生神经源性炎症反应,可以作为慢性偏头痛动物实验模型。     

鸡中枢淋巴器官肥大细胞的组织化学与形态学

对哺乳动物的,特别是啮齿动物和人类肥大细胞已有了比较深入的研究, 但关于家禽肥大细胞的研究很少.本研究旨在阐明鸡中枢淋巴器官中肥大细胞的组织化学与形态学特征.本研究证实Carnoy 氏液是鸡肥大细胞的优良的固定液,而中性缓冲福尔马林(NBF) 却阻断了大多数肥大细胞的着染力.甲苯胺蓝和阿尔新蓝是鸡肥大细胞的良好的染料,但阿尔新蓝能使更多的肥大细胞着染,虽然其也可使杯状细胞着染.作者的一种新的染色法, 长时间阿尔新蓝染色(LAB-S)可用于NBF固定的组织中肥大细胞的染色,因为其着染的细胞数与Carnoy 氏液固定甲苯胺蓝染色的细胞数无显著差异(P<0.001).在胸腺髓质中见有大量的肥大细胞,而胸腺皮质仅可见个别肥大细胞位于血管周围及小叶间结缔组织中.腔上囊的皮质与髓质中很少见有肥大细胞.肥大细胞有血管周围分布的倾向,但一个有趣的发现是血管内偶尔也有个别肥大细胞.电镜下可见肥大细胞的胞浆颗粒内充满无定形的颗粒状基质,但其电子密度有的较高,有的较低.少数胞浆颗粒内有旋涡状及网状亚微结构.但未见有人类肥大细胞胞浆颗粒内特征性的晶格状和卷轴状的亚微结构,也未见到在绵羊肥大细胞中描述过的特殊亚微结构.     

动情周期中大鼠子宫和输卵管壁肥大细胞数量变化的研究

用放射免疫分析法对动情周期中大鼠血清雌二醇浓度进行检测;取子宫、输卵管常规石蜡切片、H－E染色,并用甲苯胺蓝染色显示肥大细胞,于光镜低倍视野下计数。结果显示：动物血清雌二醇浓度依次为：动情期（E）组＞动情前期（PE）组＞动情后期（ME）组＞动情间(DE)且,各组间差异均有显性;在子宫,肥大细胞分布于宫壁肌怪平滑肌束间的结缔组织内、近小血管处,以微血管周居多,常见单个散在,于ME子宫内膜尚偶见肥大细胞;输卵管肥大细胞局限于其外膜层内、近小血管周围,亦多散在。子宫、输卵管壁内的肥大细胞镜下呈圆形、椭圆形或略不规则形,胞浆内充满紫红色粗大颗粒,子宫肥大细胞数量依次为：ME＞DE＞PE＞E,各组间差异有生（DE与PE、PE与E,P＜0.05,余组间P＜0.01）;输卵管壁内肥大细胞数量各组间差异无显性（P＞0.05）。本尚对大鼠血清雌二醇水平波动与子宫、输卵管壁内肥大细胞数量变化的关系及其生理意义进行了讨论。     

感染日本血吸虫小鼠心房特殊颗粒的形态学观察和定量分析

小鼠感染日本血吸虫尾蚴后第２８天,右心耳心房特殊颗粒数目显著减少,且多见膜溶解、断裂,半月形排空和颗粒膜与横管膜及肌膜靠近、融合及颗粒向横管腔及肌膜下突出的现象。心肌细胞内线粒体有肿胀、基质变浅及部分嵴断裂现象。感染组小鼠右心耳心房特殊颗粒的体密度（０．０３２３±０．００２９μｍ３／μｍ３）、面数密度（０．８６４７±０．０６９２μｍ－２）、数密度（３．２３６３±０．１１１４μｍ－３）及颗粒平均直径（０．２６７１±０．０２０７μｍ）均显著小于对照组的０．０９７１±０．０１２７μｍ３／μｍ３、１．９２１±０．１１４５μｍ－２、０．２１８９±０．０８６６μｍ－３和０．３１０８±０．０１９５μｍ。本文阐述了日本血吸虫感染后,促进心房特殊颗粒释放增加。     

肝素生物学功能的研究进展

肝素(heparin)是由肥大细胞产生的内源性分子,是一种高度硫酸化的糖胺聚糖,随着肥大细胞脱颗粒与组胺等物质一起释放到细胞外基质中。因其主要与抗凝血酶III结合而增强其活性,间接发挥抗凝作用,故在临床上作为抗凝剂广泛使用。然而,肝素在体内的主要生物学功能仍不清楚。研究表明,肝素能够结合体内多种蛋白,影响许多生物信息的传递,在肥大细胞活性、炎症反应、细胞增殖分化及多种疾病中发挥重要作用。本文总结肝素研究的重要成果,对肝素的主要的生物学功能进行综述。     

从甲状腺肥大细胞探讨胎儿器官肥大细胞的差异

研究肥大细胞在人胎儿甲状腺发育中数量、分布及组化性质的改变,以探讨胎儿器官发育中肥大细胞的差异。取45例不同胎龄的人胎甲状腺石蜡切片做甲苯胺蓝染色和阿尔辛蓝－－藏红染色,并测定肥大细胞的临界电解质浓度值及进行硫酸小蘖硷荧光染色。结果显示：3月龄胎儿甲状腺内开始出现肥大细胞,数量极少,主要分布在被膜及小叶间结缔组织内,甲苯胺蓝染色肥大细胞颗粒呈淡紫蓝色,阿尔辛蓝－－藏红染色呈蓝色,临界电解质浓度值较低,硫酸小蘖硷染色未见显黄色荧乐的肥大细胞,从3月龄到足月随着胎龄增长,肥大细胞数量缓慢增多,8月龄时肥大细胞经甲苯胺蓝染色,其颗粒呈紫红色,阿尔辛蓝－－藏红染色出现少量含红色和红蓝混合染色颗粒的肥大细胞,临界电解质浓度值偏高,可见少量显黄色荧光的肥大细胞,结果表明：在人胎儿3月龄时甲状腺发育中开始出现肥大细胞,但随胎儿发育肥大细胞的组化性质改变不明显。     

Mast cell ionic channels: significance for stimulus-secretion coupling.

The activation of mast cells (MC) due to immunological stimulation causes an immediate and dramatic inflammatory response. We review current evidence indicating that the membrane permeabilities for calcium, chloride, sodium, and potassium have a significant role in the activation of these cells, and in some cases, specific ionic channels have been identified. Moreover, a number of intracellular mechanisms controlling these channels are pointed out, including different classes of G proteins, intracellular calcium, cAMP, and products of phosphoinositol breakdown. However, the interplay between factors controlling membrane conductances for different ions is not currently understood. The diversity of ionic effects on MC activation is depicted, illustrating that the ionic mechanisms of MC activation are specific for different MC types. Since nerve/mast cell interaction is a key element in the burgeoning field of neuroimmunology, we discuss the role of ionic channels as targets of neurotransmitter action in MC activation.     

Distribution, Histochemical and Enzyme Histochemical Characterization of Mast Cells in Dogs

This study describes the distribution, proteoglycan properties and protease activity of mast cells from 15 different dog organs. In beagles and mixed breed dogs, staining with Alcian Blue-Safranin O revealed mast cells in all the organs examined. However, their numbers varied and they demonstrated unique localization patterns within some of these organs. Berberine sulphate fluorescence-positive mast cells were observed in the submucosa, muscularis and serosa of the intestines, as well as the tongue and liver (within the connective tissue). Mast cells within the intestinal mucosa were negative for, or demonstrated weak, berberine sulphate staining. Heterogeneity of mast cells in terms of the proteoglycans contained within their granules was further confirmed by determination of critical electrolyte concentrations (CECs). The CECs of mast cells within the connective tissue of several organs, including the intestines (submucosal and muscularis-serosal layers) were all greater than 1.0 M. The results from CEC experiments together with berberine staining indicate that heparin was contained within their granules. Relative to the CECs of mast cells in other organs, mast cells in the intestinal mucosa exhibited lower CECs, suggesting that the proteoglycans within their granules were of lower charge density and/or molecular weight. Although mast cells were classified into two groups by proteoglycans within the granules, enzyme histochemical analysis in beagles revealed three subtypes of mast cells: chymase (MC(C)), tryptase (MC(T)) and dual positive (MC(TC)) cells. There was no correlation between the proteoglycan content and enzyme properties of the mast cell granules.     

几种肥大细胞染色方法的比较

运用ABC—爱先蓝—PAS混合染色及PAP技术对大鼠肝、胃、肠及DAB诱发的大鼠肝癌中肥大细胞进行了染色对比实验,结果表明:Carnoy及福尔马林等固定液固定的组织内肥大细胞均能被爱先蓝染色成蓝色,其染色时间不同,该种染色方法可作为一种常规的染色方法,用于确定肥大细胞在组织中的分布及数量。ABC—爱先蓝—PAS混合染色方法及PAP技术均能准确、有效地确定肥大细胞(MC)性质。ABC—爱先蓝—PAS混合染色使结缔组织肥大细胞(CTMC)呈棕褐色,周边略蓝色。粘膜肥大细胞(MMC)则呈蓝色,通过不同颜色的显示,可清楚地将CTMC与MMC区分开来。PAP方法具更高的特异性。但有关的抗体来源缺乏,因此在无特异Ⅰ抗体的情况下,ABC—爱先蓝—PAS混合染色方法亦可视为鉴别两类不同性质MC的一种较为理想的方法。     

Neutralizing endogenous IL-6 renders mast cells of the MCT type from lung, but not the MCTC type from skin and lung, susceptible to human recombinant IL-4-induced apoptosis

Human cord blood-derived mast cells undergo apoptosis upon exposure to recombinant human (rh)IL-4 and become resistant to rhIL-4-induced apoptosis when cultured in the presence of rhIL-6. The current study extends these effects of rhIL-4 to different populations of human mast cells, namely fetal liver-derived mast cells, lung-derived mast cells, and skin-derived mast cells. Endogenous production of IL-6 appears to protect fetal liver-derived mast cells and those of the MC(T) phenotype from rhIL-4-mediated apoptosis, because neutralization of IL-6 renders these mast cells sensitive. In contrast, mast cells of the MC(TC) phenotype from skin and lung were resistant to IL-4-mediated apoptosis, even after neutralization of endogenous IL-6. MC(TC) cells were CD124(low), whereas those of the MC(T) cells were CD124(high). These observations extend the phenotypic differences between MC(T) and MC(TC) types of human mast cells to include different functional responses to IL-4.     

The role of mast cells in the development of skin fibrosis in tight-skin mutant mice

Chronic inflammatory conditions can evolve a fibrotic phenotype often associated with an increase in the number of mast cells (MC) near or within the granulation tissue. Despite the potential of MC to mediate fibrosis, it is unclear whether these cells play a central role in the pathogenesis of ibrosis or whether their presence is simply circumstantial. The tight-skin (Tsk) mouse develops an inherited fibrotic disease (sharing many similarties with the human disease scleroderma, systemic sclerosis) in which the lesions are associated with increased numbers and heightened granule release implicating MC in the pathogenesis of fibrosis. Despite their close association with the skin fibrosis of Tsk mice, the precise role of the MC in the pathogenesis of this inherited disease is unknown. Therefore, to assess directly whether MC are key elements in the pathogenesis of Tsk fibrosis, we generated MC deficient mice carrying the Tsk locus by utilizing selective interbreeding between Tsk and mutant mice deficient in mast cells (W, dominant white-spotting). We found that in the absence of MC, the early natural history of Tsk fibrosis was not altered. Furthermore, in older (5–7 months) Tsk mice, we found that the number of cutaneous MC was correlated with a more pronounced fibrosis. Therefore, we conclude that Tsk skin lesions are a pleiotropic manifestation of the Tsk gene in which MC are ivolved/recruited by an uncharacterized mechanism and that subsequent proliferation and activation of MC leads to augmentation of fibrosis.     

肥大细胞在肿瘤发生发展中的作用

肥大细胞是人体主要免疫细胞之一,因其作为导致过敏反应发生的最直接效应细胞而著称.肥大细胞最主要的结构特征为其胞内含有大量嗜碱性颗粒,该颗粒内又富含种类众多的生物活性物质,包括组胺、血管内皮生长因子(vascular endothelial growth factor,VEGF)、成纤维细胞生长因子(fibroblast...     

Mast cell responses to Hymenolepis microstoma infection in mice

Murine mast cells (MC) responded strongly to Hymenolepis microstoma infection. Starting day 7 postinfection (PI) and continuing until the end of the experiment (35 days PI), significantly larger numbers of MC were present in both the duodenum and bile duct of infected mice than in uninfected controls. In animals challenged with 5 cysticercoids 2 wk after primary infection, the MC response in the duodenum, but not in the bile duct, was of even greater degree than in naive hosts. The majority of MC in the duodenum of infected and challenged mice were intraepithelial mucosal MC, whereas in the bile duct the majority were connective tissue MC. Hypertrophy of the duodenal submucosa and of the bile duct wall was noticeable in all infected and challenged hosts. Worms in primary infections were not affected by the host response, but none of the worms in the challenge dose became established. It is postulated that the type of MC involved in specific immune response of the host is the intraepithelial MC, whereas the cell type participating in general inflammatory events is the connective tissue MC.     

Mastocytosis in children: clinicopathological study based on 35 cases

Immunohistochemical staining is useful in the diagnosis of bone marrow infiltration in systemic mastocytosis. However, it is not clear if antibody staining may be helpful in the diagnosis of cutaneous mastocytosis (CM). We studied the histological appearance of CM in 35 pediatric patients. Cases were assigned to three basic clinical groups: I--Urticaria pigmentosa (UP, n=29); II--Mastocytomas (n=4); and III--Diffuse Cutaneous Mastocytosis (DCM, n=2). The analysis of clinical information revealed an association between the presence of diarrhea and a higher number of cells/field. Nine doubtful cases, all of them macules, were selected based on the scarcity of mast cells (MC) and the absence or rarity of other inflammatory cells. We compared the number of cells identified in Giemsa and immunohistochemical stains in definite and doubtful cases. The intraclass correlation statistic tested the concordance between each staining method. All 9 dubious cases according to the Giemsa stain had their CM diagnosis confirmed by the immunohistochemistry analysis. The intraclass correlation between Giemsa and c-kit was good (0.7) when the number of MC was high. However, there was no correlation between the mast cells counts in the two different stains in the dubious cases. The immunohistochemistry with c-kit might make CM diagnosis easier, especially in the macular cases, when there is a lower number of MC.     

Participation of mast cells in chronic otitis media

In the pathogenesis of chronic otitis media (COM), much attention is paid to the molecular mechanisms of local inflammatory reactions in which mast cells (MCs) may be involved due to their role not only in allergic but also inflammatory processes. The aim of this study was to assess the density of mast cells in chronic otitis media in relationship to different clinical courses of COM, bacterial infections and types of disease. The MCs expression was measured immunohistochemically in paraffin-embedded granulation tissue specimens taken during surgery, by staining with a monoclonal antibody against tryptase. The density of tryptase-positive mast cells was lower in tissue samples from the group with a good clinical course than in those from the group with poor healing and recurrence (p = 0.006). There were no differences between the groups of patients with granulomatous and cholesteatomatous chronic otitis media (p = 0.66) or between the groups of patients with and without bacterial infection (p = 0.30), although the density of mast cells was lower for those with Pseudomonas aeruginosa/Proteus sp./ /Staphylococcus MRSA infection. In conclusion, the expression of mast cells in chronic otitis media granulation tissue was found to differ depending on the clinical course of the disease, but not on bacterial infection or type of COM. This may suggest that mast cells contribute to the maintenance of the inflammatory process, but not to antibacterial defense in chronic otitis media.     

流式细胞仪分析癌细胞对肥大细胞募集的影响

目的：研究食管癌细胞迁移到小鼠腹腔时肿瘤细胞周边微环境的变化。方法：将食管癌细胞株EC109和/或诱导试剂植入小鼠腹腔,利用组织化学的方法、荧光标记的肥大细胞蛋白酶和流式细胞术,我们在小鼠模型观察肠组织的形态和腹腔的MC亚型的变化。结果：胰酶导致小鼠肠道平滑肌层和黏膜下层的组织增厚,细胞间隙的增加可能有益于MC在组织移动或迁移进入腹腔;它引起小鼠腹腔液的总MC增加,MCC亚型的相对比例增加,MCT亚型减少。EC109细胞不能明显地改变小鼠肠道组织的形态,但它显著地引起腹腔MCT亚型的相对比例增加。结论：根据肥大细胞内颗粒的类胰蛋白酶和类糜蛋白酶的差异表达,可证实小鼠的MC亚型;并且不同的诱导物可能影响腹部微环境的变化。目前的研究表明,食道癌细胞可以诱导MCT（含有类胰蛋白酶）亚型迁移到小鼠腹腔,造成肿瘤细胞周围的内部环境的变化。