Survivin蛋白在大鼠急性出血坏死性胰腺炎中的表达及作用

发布时间：2019-06-17 21:22

【摘要】：目的：通过人为诱导建立大鼠急性出血坏死性胰腺炎（ANP）动物实验模型，动态观察胰腺损伤的各项指标，并检测凋亡抑制蛋白Survivin的表达在ANP中的表达及作用，从而揭示Survivin蛋白在ANP发病机制中的作用，为临床ANP诊断和治疗提供新的有效的靶点。 方法：40只健康雄性SD大鼠随机分为ANP造模后6h、12h、24h和假手术组(SO组)共4组，每组10只。手术前禁食12h，自由饮水。实验组胰胆管内推注4.5%牛磺胆酸钠1.0ml/Kg，SO组注射等量生理盐水。取各不同时间点处死SD大鼠，从心脏采血测定血淀粉酶（AMY）、血脂肪酶（LPL）的浓度。各组SD大鼠处死时取胰头部胰腺组织,应用免疫组化及RT-PCR方法检测Survivin蛋白在胰腺组织中的表达，TUNEL法检测凋亡的胰腺组织细胞的凋亡指数。 结果：假手术组（SO组）大鼠胰腺组织结构清楚，细胞形态正常，ANP实验组胰腺组织充血水肿明显，胰腺局部出现坏死灶，少量淡红色血性腹水，胰周脂肪组织皂化、出血。血清AMY于造模后6h即升高，12h左右达峰值，以后逐渐下降，但仍高于假手术组。SO组：(1276±213)U/L；ANP组(6343±287)U/L。两组总体均数间差别有统计学意义（P0.05）。大鼠ANP建立6h血清LPL即明显下降，随胰腺炎症程度的加重，血清LPL值呈逐渐下降趋势，各实验组与假手术组相比较差别有统计学意义（P 0.01）。免疫组化：假手术组（SO组）胰腺组织细胞中未见有Survivin蛋白表达，造模后ANP实验组Survivin蛋白的表达在胰腺组织细胞中主要集中于胰腺腺泡和导管细胞，呈棕褐色细颗粒沙状，ANP6h、12h、24h组Survivin的光密度值积分分别是5743±459、9041±893、13022±803，通过t-检验各组间比较差别有统计学意义（P 0.05）。RT-PCR：假手术组胰腺组织中未见Survivin蛋白mRNA的表达，大鼠ANP造模后6h即可见到Survivin蛋白mRNA的表达，并且随ANP病程的进展，Survivin蛋白mRNA的表达水平呈现逐渐增强的趋势。胰腺组织凋亡细胞呈棕黄色，假手术组胰腺组织细胞凋亡极少。ANP造模后，随胰腺炎症严重程度的加重，胰腺腺泡细胞凋亡指数逐渐降低，与假手术组相比较差别有显著统计学意义（P 0.01），胰腺导管细胞凋亡指数逐渐升高，与假手术组相比较差别有统计学意义（P 0.05）。 结论：1.大鼠ANP动物实验随病程延长，胰腺组织细胞损伤程度呈逐渐加重趋势。2.血淀粉酶、血脂肪酶对ANP诊断有着重要的意义，但对于评估ANP损伤严重程度无重要参考意义。3.Survivin在ANP中的表达显著增加，Survivin表达增加与ANP损伤程度有密切关系，，对于判断ANP预后有重要意义。
[Abstract]:Aim: to establish an animal model of acute hemorrhagic necrotizing pancreatitis (ANP) in rats by artificial induction, to observe the indexes of pancreatic injury, and to detect the expression and role of apoptosis inhibitor protein Survivin in ANP, so as to reveal the role of Survivin protein in the pathogenesis of ANP and provide a new and effective target for clinical diagnosis and treatment of ANP. Methods: forty healthy male SD rats were randomly divided into 4 groups at 6 h, 12 h, 24 h and SO group (SO group) with 10 rats in each group. Fasting for 12 hours before operation, free drinking water. 4.5% sodium taurocholate 1.0 ml 路kg ~ (- 1) was injected into the pancreatic bile duct of the experimental group, and the same amount of saline was injected into the so group. SD rats were killed at different time points, and the concentration of serum amylase (AMY), blood lipase (LPL) was measured from the heart. The pancreatic tissue of pancreatic head was taken at the time of execution of SD rats in each group. The expression of Survivin protein in pancreatic tissue was detected by immunohistochemistry and RT-PCR, and the apoptosis index of apoptotic pancreatic tissue was detected by TUNEL assay. Results: in the sham operation group (SO group), the structure of pancreatic tissue was clear and the morphology of cells was normal. The hyperemia and edema of pancreatic tissue were obvious in ANP experimental group, and the local necrotic foci, a small amount of reddish hemorrhagic ascitic fluid, saponification and bleeding of peripancreatic adipose tissue were found in ANP experimental group. Serum AMY increased at 6 h after modeling, reached its peak at 12 h, and then decreased gradually, but it was still higher than that in pseudo-operation group. SO group: (1276 卤213) U 鈮

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