胆囊收缩素、胰岛素与胰高血糖素分泌细胞在小鼠胰腺发育过程中的形态学观察

发布时间：2018-05-11 15:41

本文选题：小鼠胰腺 + 发育　；参考：《山西医科大学》2009年硕士论文

【摘要】： 目的: 观察发育中昆明小鼠胰腺内胆囊收缩素免疫反应细胞(cholecystokinin immunoreactive cells, CCK-IR细胞)、胰岛素免疫反应细胞(insulin immunoreactive cells, Ins-IR细胞)和胰高血糖素免疫反应细胞(glucagon immunoreactive cells, Glu-IR细胞)的发生、分布和形态特征;以及在胰腺内胆囊收缩素与胰岛素,胰岛素与胰高血糖素是否有在同一细胞中共存的现象。方法: 应用免疫组织化学SABC法观察小鼠胚胎第11天到出生后第45天胰腺内CCK-IR细胞、Ins-IR细胞和Glu-IR细胞的发生、分布及形态特征。并用免疫荧光双标法观察胰腺内胆囊收缩素与胰岛素、胰岛素与胰高血糖素在同一细胞中是否有共存的现象。结果: 免疫组织化学SABC法显示,小鼠胚胎第11天,胰腺尚处于分化初期,在胰腺内开始出现CCK-IR细胞、Ins-IR细胞和Glu-IR细胞。3种细胞常成簇分布,浅染,细胞界限不清。至胚胎第15天后,3种细胞着色深浅不一,细胞界限逐渐可辨,在胰岛内呈散在分布。胚胎晚期至出生后,深染的Ins-IR细胞和Glu-IR细胞数量增多,Ins-IR细胞在胰岛内渐呈弥散状分布,而Glu-IR细胞在胰岛内由散在分布逐渐移向周边。出生后在胰岛内仅偶可见浅染的CCK-IR细胞。胰腺外分泌部的导管和腺泡上皮细胞之间,胚胎期偶可见到上述3种细胞,但小鼠出生后仅可见到Ins-IR细胞和Glu-IR细胞,而未见CCK-IR细胞。免疫荧光法显示,CCK-IR细胞的形态多样,有些细胞可见伸出的细长突起。在胚胎期,CCK-IR细胞的荧光强弱不一,出生后,荧光逐渐减弱,但在出生后第45天,胰岛的边缘部仍可见到CCK-IR细胞。Ins-IR细胞和Glu-IR细胞的形态以椭圆形、圆形多见。胚胎第15天前,Ins-IR细胞和Glu-IR细胞的荧光较弱,其中,Glu-IR细胞的荧光强度比Ins-IR细胞强。胚胎晚期直至出生后第45天,两种细胞的荧光都逐渐增强。免疫荧光双标法观察,自胚胎第14天至出生后第45天,小鼠胰腺内始终可见CCK-Ins-IR细胞,其数量以胚胎晚期和生后第30天较多。自胚胎第11天至出生后第45天,小鼠胰腺内可见Ins-Glu-IR细胞,其数量以胚胎第11天和生后第30天最多。生后在胰腺的外分泌部中也偶然可见Ins-Glu-IR细胞。结论: 1.对于显示发育中胰腺内的CCK-IR细胞,免疫荧光法比普通免疫组织化学技术更为敏感。 2.发育中小鼠胰腺内CCK-IR细胞具有较典型的旁分泌细胞的形态特征;该细胞的分布时段提示,CCK-IR细胞可能以内分泌、旁分泌等方式影响胰岛内分泌细胞及胰腺外分泌部的发生、发育,并参与调节小鼠生后胰腺的分泌活动。 3.免疫荧光双标法可以较好地显示两种激素在同一细胞内的共存现象,本实验中所显示的CCK-Ins-IR细胞、Ins-Glu-IR细胞可能是胰腺内较幼稚的内分泌细胞。 4.小鼠生后第30天的胰岛内可见到大量Ins-Glu-IR细胞,据此推测,在发育成熟的小鼠胰岛内的B细胞仍可能具有分裂增殖的潜能。
[Abstract]:Objective: The occurrence, distribution and morphological characteristics of cholecystokinin immunoreactive cells, CCK-IR cells, insulin immunoreactive cells (Ins-IR cells) and glucagon immunoreactive cells (Glu-IR cells) in the pancreas of developing Kunming mice were observed. And whether cholecystokinin and insulin, insulin and glucagon coexist in the same cell in the pancreas. Methods: The occurrence, distribution and morphological characteristics of CCK-IR cells, Ins-IR cells and Glu-IR cells in the pancreas of mice from the 11th day to the 45th day after birth were observed by immunohistochemical SABC method. The co-existence of cholecystokinin and insulin, insulin and glucagon in the same cell were observed by immunofluorescence double labeling method. Results: Immunohistochemical SABC showed that the pancreas was still in the early stage of differentiation on the 11th day of embryo. CCK-IR cells, Ins-IR cells and Glu-IR cells began to form clusters, light staining and unclear cell boundaries. After the 15th day of embryo, the three kinds of cells were stained in different depth, the cell boundaries were gradually discernible and dispersed in the islets. From late embryonic stage to postnatal stage, the number of Ins-IR cells and Glu-IR cells with deep staining increased. Ins-IR cells gradually distributed in the islets, while Glu-IR cells gradually moved from the scattered distribution to the periphery in the islets. Only slightly stained CCK-IR cells were seen in the islets after birth. Between the ducts and acinar epithelial cells in the exocrine part of the pancreas, the above three types of cells were occasionally found in the embryonic phase, but only Ins-IR cells and Glu-IR cells were found in mice after birth, but no CCK-IR cells were found. Immunofluorescence assay showed that CCK-IR cells had a variety of morphologies, and some cells had elongated protuberances. The fluorescence intensity of CCK-IR cells was different in embryonic stage, but the fluorescence decreased gradually after birth, but on the 45th day after birth, the morphology of CCK-IR cells. Ins-IR cells and Glu-IR cells were oval and round. The fluorescence intensity of Glu-IR cells was stronger than that of Ins-IR cells. The fluorescence of both cells increased gradually in late embryo and 45 days after birth. From the 14th day of embryo to the 45th day after birth, CCK-Ins-IR cells were observed in the pancreas of mice by immunofluorescence double labeling method. The number of CCK-Ins-IR cells was more in late embryo and 30 days after birth. From the 11th day of embryo to the 45th day after birth, Ins-Glu-IR cells could be seen in the pancreas of mice. The number of Ins-Glu-IR cells was the highest on the 11th day of embryo and the 30th day after birth. Ins-Glu-IR cells are also occasionally found in the exocrine portion of the pancreas after birth. Conclusion: 1. Immunofluorescence method is more sensitive than ordinary immunohistochemical technique to display CCK-IR cells in developing pancreas. 2. CCK-IR cells in the pancreas of developing mice have typical paracrine cells, and the distribution period of the cells suggests that CCK-IR cells may affect the development of islet endocrine cells and pancreatic exocrine cells by endocrine and paracrine methods. And involved in regulating the secretion of mouse pancreas after birth. 3. Immunofluorescence double labeling method can well show the coexistence of two hormones in the same cell. The CCK-Ins-IR cells in this study may be immature endocrine cells in the pancreas. 4. On the 30th day after birth, a large number of Ins-Glu-IR cells could be seen in the islets of mice, so it was speculated that the B cells in the mature islets of mice might still have the potential to divide and proliferate.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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