本文選題：小鼠胰腺 + 發(fā)育��； 參考：《山西醫(yī)科大學(xué)》2009年碩士論文

【摘要】： 目的: 觀察發(fā)育中昆明小鼠胰腺內(nèi)膽囊收縮素免疫反應(yīng)細(xì)胞(cholecystokinin immunoreactive cells, CCK-IR細(xì)胞)、胰島素免疫反應(yīng)細(xì)胞(insulin immunoreactive cells, Ins-IR細(xì)胞)和胰高血糖素免疫反應(yīng)細(xì)胞(glucagon immunoreactive cells, Glu-IR細(xì)胞)的發(fā)生、分布和形態(tài)特征;以及在胰腺內(nèi)膽囊收縮素與胰島素,胰島素與胰高血糖素是否有在同一細(xì)胞中共存的現(xiàn)象。 方法: 應(yīng)用免疫組織化學(xué)SABC法觀察小鼠胚胎第11天到出生后第45天胰腺內(nèi)CCK-IR細(xì)胞、Ins-IR細(xì)胞和Glu-IR細(xì)胞的發(fā)生、分布及形態(tài)特征。并用免疫熒光雙標(biāo)法觀察胰腺內(nèi)膽囊收縮素與胰島素、胰島素與胰高血糖素在同一細(xì)胞中是否有共存的現(xiàn)象。 結(jié)果: 免疫組織化學(xué)SABC法顯示,小鼠胚胎第11天,胰腺尚處于分化初期,在胰腺內(nèi)開始出現(xiàn)CCK-IR細(xì)胞、Ins-IR細(xì)胞和Glu-IR細(xì)胞。3種細(xì)胞常成簇分布,淺染,細(xì)胞界限不清。至胚胎第15天后,3種細(xì)胞著色深淺不一,細(xì)胞界限逐漸可辨,在胰島內(nèi)呈散在分布。胚胎晚期至出生后,深染的Ins-IR細(xì)胞和Glu-IR細(xì)胞數(shù)量增多,Ins-IR細(xì)胞在胰島內(nèi)漸呈彌散狀分布,而Glu-IR細(xì)胞在胰島內(nèi)由散在分布逐漸移向周邊。出生后在胰島內(nèi)僅偶可見淺染的CCK-IR細(xì)胞。 胰腺外分泌部的導(dǎo)管和腺泡上皮細(xì)胞之間,胚胎期偶可見到上述3種細(xì)胞,但小鼠出生后僅可見到Ins-IR細(xì)胞和Glu-IR細(xì)胞,而未見CCK-IR細(xì)胞。 免疫熒光法顯示,CCK-IR細(xì)胞的形態(tài)多樣,有些細(xì)胞可見伸出的細(xì)長(zhǎng)突起。在胚胎期,CCK-IR細(xì)胞的熒光強(qiáng)弱不一,出生后,熒光逐漸減弱,但在出生后第45天,胰島的邊緣部仍可見到CCK-IR細(xì)胞。Ins-IR細(xì)胞和Glu-IR細(xì)胞的形態(tài)以橢圓形、圓形多見。胚胎第15天前,Ins-IR細(xì)胞和Glu-IR細(xì)胞的熒光較弱,其中,Glu-IR細(xì)胞的熒光強(qiáng)度比Ins-IR細(xì)胞強(qiáng)。胚胎晚期直至出生后第45天,兩種細(xì)胞的熒光都逐漸增強(qiáng)。 免疫熒光雙標(biāo)法觀察,自胚胎第14天至出生后第45天,小鼠胰腺內(nèi)始終可見CCK-Ins-IR細(xì)胞,其數(shù)量以胚胎晚期和生后第30天較多。自胚胎第11天至出生后第45天,小鼠胰腺內(nèi)可見Ins-Glu-IR細(xì)胞,其數(shù)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見Ins-Glu-IR細(xì)胞。 結(jié)論: 1.對(duì)于顯示發(fā)育中胰腺內(nèi)的CCK-IR細(xì)胞,免疫熒光法比普通免疫組織化學(xué)技術(shù)更為敏感。 2.發(fā)育中小鼠胰腺內(nèi)CCK-IR細(xì)胞具有較典型的旁分泌細(xì)胞的形態(tài)特征;該細(xì)胞的分布時(shí)段提示,CCK-IR細(xì)胞可能以內(nèi)分泌、旁分泌等方式影響胰島內(nèi)分泌細(xì)胞及胰腺外分泌部的發(fā)生、發(fā)育,并參與調(diào)節(jié)小鼠生后胰腺的分泌活動(dòng)。 3.免疫熒光雙標(biāo)法可以較好地顯示兩種激素在同一細(xì)胞內(nèi)的共存現(xiàn)象,本實(shí)驗(yàn)中所顯示的CCK-Ins-IR細(xì)胞、Ins-Glu-IR細(xì)胞可能是胰腺內(nèi)較幼稚的內(nèi)分泌細(xì)胞。 4.小鼠生后第30天的胰島內(nèi)可見到大量Ins-Glu-IR細(xì)胞,據(jù)此推測(cè),在發(fā)育成熟的小鼠胰島內(nèi)的B細(xì)胞仍可能具有分裂增殖的潛能。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

【共引文獻(xiàn)】

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