葡萄糖调节蛋白与重症急性胰腺炎中性粒细胞凋亡关系研究

发布时间：2019-05-15 05:41

【摘要】：目的：探讨葡萄糖调节蛋白78（GRP78）在牛黄胆酸钠诱导的大鼠重症急性胰腺炎时对中性粒细胞凋亡的作用。 方法：将54只雄性SD大鼠随机分为两组，分别建立假手术组（SO）及重症急性胰腺炎组（SAP）模型，SO组24只、SAP组30只，5％的牛黄胆酸钠逆行胰胆管注射制备大鼠重症急性胰腺炎模型，对照组只开腹翻动胰腺即缝合，造模后分别于3h,6h,12h观察大鼠的一般情况及死亡率，并处死大鼠，抽取静脉血测定各个时间点血清淀粉酶的浓度，分离中性粒细胞并用流式细胞仪检测中性粒细胞的凋亡率，提取中性粒细胞蛋白做western-blot测定GRP78,Mcl-1蛋白的表达，用HE染色切片观察胰腺的病理变化并对胰腺组织进行病理评分。 结果：重症急性胰腺炎组的血清淀粉酶的浓度明显高于对照组(P0.01)，SAP组在3h，6h，12h的淀粉酶浓度逐渐升高(P0.01)，至术后12h升至（6246.00±602.931）；SAP组中性粒细胞凋亡率较SO组明显降低，随时间延长而下降(P0.01)，至术后12h降至（1.300±0.5812）；western-blot显示SAP组中性粒细胞中GRP78,Mcl-1的表达较SO组明显升高，随着时间延长更明显；SAP大鼠PMN的凋亡率与GRP78,Mcl-1蛋白水平呈负相关（r=-0.836，，P0.01）、（r=-0.895，P0.01）；病理HE染色SAP组有大量炎细胞及红细胞浸润。 结论:本研究表明，大鼠重症胰腺炎时，外周血中性粒细胞的凋亡率随时间延长而下降，GRP78的表达明显增高，这可能与应激产生的GRP78的表达增强激活PI3k/Akt信号通路从而激活Mcl-1，在外周血中性粒细胞的凋亡延迟中发挥作用。
[Abstract]:Aim: to investigate the effect of glucose regulated protein 78 (GRP78) on neutrophil apoptosis in rats with severe acute pancreatitis induced by sodium taurocholate. Methods: 54 male SD rats were randomly divided into two groups. (SO) model was established in sham operation group and (SAP) model in severe acute pancreatitis group. There were 24 rats in SO group and 30 rats in SAP group. The rat model of severe acute pancreatitis was established by retrograde injection of 5% sodium taurocholate into the pancreatic duct. In the control group, the pancreas was sutured only by laparotomy. The general situation and mortality of the rats were observed at 3 h and 6 h, respectively, and the rats were killed. Venous blood was taken to determine the concentration of serum amylase at each time point, neutrophils were isolated and flow cytometry was used to detect the apoptosis rate of neutrophils. Neutrophil proteins were extracted and the expression of GRP78,Mcl-1 protein was detected by western-blot. The pathological changes of pancreas were observed by HE staining and the pathological scores of pancreatic tissue were evaluated. Results: the concentration of serum amylase in severe acute pancreatitis group was significantly higher than that in control group (P 0.01), SAP group at 3 h, 6 h, 12 h, P 0.01), and increased to (6246.00 卤602.931) at 12 h after operation. The apoptosis rate of neutrophils in SAP group was significantly lower than that in SO group, which decreased with the prolongation of time (P 0.01) and decreased to (1.300 卤0.5812) at 12 h after operation. Western-blot showed that the expression of GRP78,Mcl-1 in neutrophils of SAP group was significantly higher than that of SO group, especially with the prolongation of time. The apoptosis rate of PMN in SAP rats was negatively correlated with the level of GRP78,Mcl-1 protein (r 鈮

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