【摘要】：目的：探討葡萄糖調(diào)節(jié)蛋白78（GRP78）在牛黃膽酸鈉誘導(dǎo)的大鼠重癥急性胰腺炎時對中性粒細(xì)胞凋亡的作用。 方法：將54只雄性SD大鼠隨機(jī)分為兩組，分別建立假手術(shù)組（SO）及重癥急性胰腺炎組（SAP）模型，SO組24只、SAP組30只，5％的牛黃膽酸鈉逆行胰膽管注射制備大鼠重癥急性胰腺炎模型，對照組只開腹翻動胰腺即縫合，造模后分別于3h,6h,12h觀察大鼠的一般情況及死亡率，并處死大鼠，抽取靜脈血測定各個時間點(diǎn)血清淀粉酶的濃度，分離中性粒細(xì)胞并用流式細(xì)胞儀檢測中性粒細(xì)胞的凋亡率，提取中性粒細(xì)胞蛋白做western-blot測定GRP78,Mcl-1蛋白的表達(dá)，用HE染色切片觀察胰腺的病理變化并對胰腺組織進(jìn)行病理評分。 結(jié)果：重癥急性胰腺炎組的血清淀粉酶的濃度明顯高于對照組(P0.01)，SAP組在3h，6h，12h的淀粉酶濃度逐漸升高(P0.01)，至術(shù)后12h升至（6246.00±602.931）；SAP組中性粒細(xì)胞凋亡率較SO組明顯降低，隨時間延長而下降(P0.01)，至術(shù)后12h降至（1.300±0.5812）；western-blot顯示SAP組中性粒細(xì)胞中GRP78,Mcl-1的表達(dá)較SO組明顯升高，隨著時間延長更明顯；SAP大鼠PMN的凋亡率與GRP78,Mcl-1蛋白水平呈負(fù)相關(guān)（r=-0.836，，P0.01）、（r=-0.895，P0.01）；病理HE染色SAP組有大量炎細(xì)胞及紅細(xì)胞浸潤。 結(jié)論:本研究表明，大鼠重癥胰腺炎時，外周血中性粒細(xì)胞的凋亡率隨時間延長而下降，GRP78的表達(dá)明顯增高，這可能與應(yīng)激產(chǎn)生的GRP78的表達(dá)增強(qiáng)激活PI3k/Akt信號通路從而激活Mcl-1，在外周血中性粒細(xì)胞的凋亡延遲中發(fā)揮作用。
[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 鹵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 鹵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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