双肾上腺皮质激素样激酶-1和Ki-67在胰管结扎后胰岛再生中的表达及分布

发布时间：2018-04-25 23:32

本文选题：双肾上腺皮质激素样激酶1 + Ki-67　；参考：《大理大学》2017年硕士论文

【摘要】：目的:研究胰管结扎(Pancreatic duct ligation,PDL)术后双肾上腺皮质激素样激酶-1(Doublecortin and Ca M kinase-like-1,DCLK1)和增殖细胞核抗原(Nuclcar-associated antigen,Ki-67)在胰岛再生中的表达及分布,为探讨胰岛再生的分子调控机制提供实验依据。方法:随机将35只大鼠(体质量250±10.8g)分为正常对照组(5只)和胰管结扎(PDL)模型组(30只),模型组再分成6个亚组(每组为5只)。模型组采用4%的水合氯醛腹腔麻醉后于胰尾总管处结扎主胰管并于术后1 d、3d、5d、7d、9d、12d分别取胰腺(体、尾部),与对照组(胰腺体、尾部)进行组织学处理。模型组与对照组均采用免疫组织化学SABC染色法观察DCLK1和Ki-67在胰腺组织中的表达及分布,并通过H-E染色,Insulin(Ins)及Glucagon(Glu)免疫染色观察胰岛的再生情况。结果:DCLK1和Ki-67在正常胰腺组织中存在少量表达,其表达主要分布在胰腺导管、腺泡和胰岛。PDL术后1、3天,胰腺组织高度水肿,腺泡细胞变性坏死,胰岛形态被破坏,胰岛内β细胞变性坏死,胰岛边缘α细胞呈Glu免疫阳性反应;DCLK1在胰腺组织内呈现异常表达;Ki-67在第3天可见少量免疫阳性细胞,其分布无特殊。PDL术后5天,坏死的胰腺组织脂肪化,胰腺小叶内的大量腺泡呈脂肪样变,胰腺小叶内开始出现管状复合体(tubular complexes,TC)结构,DCLK1在管状复合体上皮呈较强的免疫阳性反应,Ki-67阳性细胞数量增多较为明显。PDL术后7、9天,外分泌部萎缩,许多胰腺小叶被管状复合体结构及脂肪组织替代,DCLK1表达减弱,胰岛内开始出现Ins免疫阳性反应。PDL术后12天,胰腺腺泡结构恢复,外分泌部之间出现正常胰岛,Ins、Glu免疫阳性反应较明显,DCLK1未见表达,Ki-67阳性细胞数恒定,分布清晰,主要分布在导管、腺泡及胰岛,分布具有差异性,差异具有统计学意义(P0.05)。结论:胰管结扎后,早期胰腺外分泌部组织呈脂肪化,DCLK1异常表达于TC结构;后期DCLK1表达下降,胰岛细胞形态及内分泌功能恢复正常。Ki-67阳性细胞分布以导管上皮最为明显,其次是腺泡及胰岛,分布具有差异性。
[Abstract]:Objective: to investigate the expression and distribution of double adrenocorticortin and Ca M kinase-like kinase-1 (DCLK1) and proliferating cell nuclear antigen Nuclcar-associated antigen Ki-67 (PCNA) in islet regeneration after pancreatic duct ligation with Pancreatic duct ligation (PDL). Methods: 35 rats (250 卤10.8g body weight) were randomly divided into normal control group (n = 5) and pancreatic duct ligation model group (n = 30). The model group was divided into 6 subgroups (5 rats in each group). In the model group, the main pancreatic duct was ligated at the common pancreatic duct after abdominal anesthesia with 4% chloral hydrate, and the pancreas (body and tail) were removed at 1 day, 3 days, 5 days, 7 days and 12 days, respectively, and the control group (pancreatic body, tail) were treated with histology. The expression and distribution of DCLK1 and Ki-67 in pancreatic tissues were observed by immunohistochemical SABC staining in both the model group and the control group. The regeneration of pancreatic islets was observed by H-E staining and Glu-staining. Results there was a small amount of expression of 7% DCLK1 and Ki-67 in normal pancreatic tissues. The expression was mainly distributed in pancreatic ducts, acinar and islet. PDL showed high edema, degeneration and necrosis of acinar cells and destruction of pancreatic islets. 尾 -cell degeneration and necrosis in the islet, Glu immunoreactivity of 伪 cells at the edge of the islet. A small number of immunoreactive cells were found in the pancreatic tissue on the 3rd day, and there was no special expression of Ki-67 on the 5th day after the operation. The necrotic tissue of the pancreas is fatty, and a large number of acinus in the lobules of the pancreas show fatty degeneration. In the pancreatic lobule, the tubular complex (tubular complex TC1) structure began to appear. DCLK1 showed a stronger immunoreactive reaction in the tubular complex epithelium. The number of Ki-67 positive cells increased obviously. The exocrine part shrank 7 days after PDL, and the exocrine part was atrophied at 7 ~ 9 days after PDL. In many pancreatic lobules, the expression of DCLK1 was weakened by tubular complex structure and adipose tissue replacement. The pancreatic acinar structure recovered 12 days after the onset of Ins immunoreactivity in pancreatic islets. The positive expression of Ki-67 was not found in DCLK1. The expression of Ki-67 was distinct in the ducts, acinar and islets. The distribution was different, and the difference was statistically significant (P 0.05). Conclusion: after ligation of pancreatic duct, the tissue of pancreatic exocrine showed abnormal expression of DCLK1 in TC structure in the early stage, the expression of DCLK1 decreased in the later stage, and the morphology and endocrine function of pancreatic islet cells returned to normal. The distribution of Ki-67 positive cells was the most obvious in ductal epithelium. The distribution of acinar and pancreatic islets was different.
【学位授予单位】：大理大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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