【作者】 卢晓晔；

【导师】 王自能；

【作者基本信息】 暨南大学 ， 妇产科学， 2003， 博士

【摘要】 弥散神经内分泌系统(diffuse neuroendocrine system，DNES)，由弥散分布于机体多种组织器官中的内分泌细胞(endocrine cell，EC)与具有内分泌功能的神经元组成。DNES细胞的共同特点是可以合成与分泌多种胺和／或肽，通过内分泌(endocrine)、旁分泌(paracrine)、神经分泌(neurocrine)和腔分泌(lumenocrine)的方式调节靶细胞的发育分化、代谢及功能活动。此外，内分泌细胞也可见于多种肿瘤组织，与肿瘤的生长、浸润、转移及血管形成等有关。 由于内分泌细胞具有亲银(argentaffin)或嗜银特性(argyrophilic)，又称嗜银细胞(argyrophil cell)。银染色法是一种传统的鉴定内分泌细胞的方法，但其敏感性及特异性较差。目前主要采用免疫组化法检测内分泌细胞，常用的特异性标记物有神经元特异性烯醇化酶(neuron-specific enolase，NSE)、嗜铬素A(chromogranin A，CgA)和突触素(synaptophysin，Syn)。CgA与多种肽类激素及神经肽共同储存于多种内分泌细胞及神经元的分泌颗粒内，参与调节多肽类激素的贮存和分泌过程。有研究表明，CgA是神经内分泌细胞的最佳标记物，其特异性和敏感性分别达85.7％和67.9％，因此能标记出绝大部分内分泌细胞。 已有的研究显示，正常子宫内膜仅存在少量内分泌细胞，而在子宫内膜癌组织内分泌细胞较常见。免疫组织化学证实，子宫内膜癌的内分泌细胞含单胺和多种活性肽，如血清素(serotonin或5-HT)、生长抑素(somatostatin)、FSH、ACTH、血管活性肠肽(vasoactive intestinal peptide，VIP)、胃泌素(gastrin)、蛙皮素厂ombesin）和降钙素k 等。尽管子宫内膜内分泌细胞的功能意义尚不清楚，但其分泌的多种胺和肽具有调节靶细胞的分裂增殖、生长分化、代谢及功能活动等多方面的作用。体内、外的研究发现，这些肽类激素的桔抗剂或类似物能抑制肿瘤生长，对治疗神经内分泌肿瘤有一定的疗效。 子宫是性激素依赖器官，雌、孕激素对子宫发育与功能活动的重要性已为人们所熟知，但随着人们对多肽类激素、生长因子、自分泌与旁分泌调节以及上皮细胞与间质相互作用的重要性的认识，子宫内膜本身存在的内分泌细胞的作用也不容忽视。有关子宫内膜内分泌细胞的研究远不及胃、肠、胰、肺、前列腺及其相应肿瘤组织中内分泌细胞的研究广泛和深入。子宫内膜内分泌细胞的来源及胚胎发育规律尚未见报道。子宫内膜内分泌细胞的生物学特性、内分泌细胞与雌、孕激素受体的关系亦鲜见报道。上述这些问题的探讨，将有助于了解内分泌细胞对子宫内膜的发育、功能活动以及对子宫内膜癌发生、发展及生物学行为的影响，并可为子宫内膜癌的合理治疗提供理论依据。 第一部分 人正常子宫内膜与子宫内膜癌 内分泌细胞的形态学研究 目的：观察正常子宫内膜及子宫内膜癌组织中内分泌细胞的光镜及超微结构特征。 方法：12－40周胎儿子宫内膜（n－20）、育龄妇女正常子宫内膜O—20）及子宫内膜样腺癌组织（n＝＝50），采用嗜铬素 A（chromogranin A，CgA）免疫组织化学染色标记内分泌细胞，光镜观察其分布及形态特点；并采用透射电镜观察内分泌细胞的超微结构特点。 2 结果： 1．胎儿子宫内膜内分泌细胞的免疫组化及超微结构观察 最早在16周胎儿子宫内膜标记出即A阳性细胞。CgA阳性细胞弥散分布于表面上皮及腺上皮，以靠近基底处较多。CgA阳性细胞的形态多样，有细长的突起，并可见突起相互连接。 由于电镜标本取材范围局限，仅在1例足月妊娠的胎儿子宫内膜发现内分泌细胞。内分泌细胞呈三角形，含有膜包的高电子密度的分泌颗粒，直径为240－480urn（平均360urn）。 2．育龄妇女正常子宫内膜内分泌细胞的免疫组化及超微结构观察 20例正常子宫内膜中有3例呈CgA阳性反应，其中增生期2例，分泌期1例。正常子宫内膜组织中CgA阳性细胞稀少，仅出现于极个别腺体上。CgA阳性反应呈细小颗粒状分布于腺上皮细胞的核下区或核上区。 2例增生期子宫内膜在电镜下发现内分泌细胞，胞质含膜包的高电子密度颗粒。还可见到一些膜性空泡状结构，空泡的表面大多呈波纹状。此外，尚可见同时含有内分泌颗粒及粘液颗粒的“双分泌细胞（amphicrine cell）。粘液颗粒有单位膜包绕，内容物为中等电子密度的细颗粒或呈Y纹状”结构。 3．子宫内膜样腺癌组织中内分泌细胞的免疫组化观察 22例子宫内膜样腺癌组织中检测到CgA阳性细胞。CgA阳性细胞常单个分散穿插于腺上皮细胞之间或相对集中分布于肿瘤的某一区域的腺体或实性巢，也可见3个以上的CgA阳性细胞成群存在于癌组织内。 单个的 CgA阳性细胞形态多样，有的 CgA阳性细胞具有神经元样的突起，延伸到比邻细胞或基底层。子宫内膜样腺癌组织中大多数内分泌细胞的CgA阳性产物充满胞浆，呈强阳性。与正常细胞相比，子宫内膜样腺癌组织中 CgA阳性细胞有不同程度的异型性，核大而不?更多还原

【Abstract】 Diffuse neuroendocrine system (DNES) includes secreting neurons and endocrine cells (EC), which diffuse distributed in many sites of the body. The common features of DNES are they can secrete various kinds of amine and/or peptides, and they modulate the development, differentiation, metabolism and function by endocrine, paracrine and autocrine, neurocrine and lumenocrine. DNES cells also present in a number of tumors, and appear to be associated with tumor growth, invasiveness, metastatic processes and angiogenesis.Endocrine cells are also called argyrophil cells because their argentaffin or argyrophilic properties. Grimelius silver nitrate staining is a traditional method to detect endocrine cells, but its sensitivity and specificity are unsatisfied. Immunohistochemical techniques are used to identify the endocrine cells nowaday. The commonly used markers are neuron-specific enolase (NSE), chromogranin A(CgA) and synaptophysin(Syn). CgA, a member of the family of acidic secretory proteins found in the secretory granules of a wide variety of endocrine cells and neurons, where it is stored together with many different peptid hormones and neuropeptides, may be involved in the packaging peptides into secretory granules and may modulate peptide hormone processing. The specificity and sensitivity of CgA was 85.7% and 67.9% respectively, therefore CgA can be used to demonstrate most of endocrine cells.Some researchs have showed that there are scanty endocrine cells in normal endometrium, while they are frequently present in endometrial carcinoma. A number of bioactive secretory productes such as serotonin, somatostatin, FSH, ACTH, vasoactive intestinal peptide, gastrin, bombesin have been identified by monoclonal antisera in endometrioid adenocarcinoma. Very little is known about the function of endocrine cells in endometrioid adenocarcinoma, however several endocrine hormones, including serotonin, bombesin and somatostatin, are known to manifest growth-modulating activity and may influence the growth, differentiation and function of the target cells. A number of analogues or agonists of the endocrine hormones have studied in vitro and in vivo, and the results showed they are effective in treating tumor-related syndromes.It is well-know that uterus as an organ is dependent on estrogen and progesterone for its normal development and function. The importance of peptide hormones, growth factors, autocrine-paracrine regulatory loops, and stromal-epithelial interactions is now widely recognized. So, the role of endocrine cells in endometrium should not be neglected. The studies on endocrine cells of endometrium are far less than that on gastrointestinal trac^ lung^ pancreas and prostate. Our investigations are try to explore the histogenesis of endometrial endocrine cells, their development during embryonic period, their biological characteristic and significance in normal and malignant endometrial tissue, and to provide a new approach of therapeutic procedures based on neuroendocrine hormones and their antagonists.Part oneMorphological Observation on Endocrine Cells of Human Normal Endometrium and Endometrioid AdenocarcinomObjective: To observe the morphological and ultrastructural characteristic of normal endometrium and endometrioid adenocarcinoma.Materials and Methods: Fetal endometrium of 12-40 week gestation (n=20), normal adult endometrium (n=20) and endometrioid adenocarcinoma (n=50) were studied by immunohistochemistry and transmission electron microscope.Results:1. Immunohistochemistry and ultrastructural observations of fetal endometriumChromogranin A positive cells appeared in 16-week fetus and located in the surface and glandular epithelium. Cells containing serotonin, somatostatin and gastrin-releasing peptide did not found.The ultrastructure of endocrine cells showed that their cytoplasm contained specific dense core vesicles.2. Immunohistochemistry and ultrastructural observations of normal endometriumScanty CgA positive cells w更多还原