急性梗阻性胰腺炎大鼠胰腺细胞凋亡及其内分泌变化的实验研究

Pancreatic cell apoptosis and endocrine alterations in rats with acute obstructive pancreatitis

目的探讨急性梗阻性胰腺炎（acute obstructive pancreatitis，AOP）大鼠胰腺细胞凋亡的发生规律及其内分泌相关激素的变化。方法采用胆胰管结扎方法建立AOP大鼠模型，分别于梗阻后8、12h取血检测胰岛素和胰高血糖素以及血清淀粉酶水平；取胰腺组织，采用TUNEL法与Annexin—Ⅴ／PI双染法流式细胞仪检测胰腺细胞的凋亡。结果AOP大鼠在胆胰管结扎8、12h时的血清淀粉酶分别为（1198±687）U／L和（1698±1103）U／L，胰岛素分别为（8．1±5．8）ng／ml和（12．7±6．9）ng／L，胰高血糖素分别为（6．8±4．6）ng／ml和（7．3±2．9）ng／ml，均显著高于假手术对照组的（404±222）U／L、（5．6±2．7）ng/ml和（2．6±2．1）ng／ml（P〈0．05）。胰腺组织中外分泌部分腺泡细胞与内分泌部分胰岛细胞均发生凋亡，8、12h的细胞凋亡率分别为（20．5±11．2）％和（15．5±8．9）％，均显著高于假手术组的（4．2±1．6）％（P〈0．05）。结论AOP大鼠的胰腺组织中内、外分泌腺细胞均发生凋亡，这可能是AOP内分泌激素改变的病理生理学基础。

Objective To investigate the occurrence of apoptosis and its relationship with endocrine hormones alterations in rats with acute obstructive pancreatitis （AOP）. Methods The model of AOP was established by ligation of panereaticobiliary duct. 8, 12 hrs after operation, the serum insulin, glucagons and amylase were determined; pancreatic tissues were harvested and apoptotic rate was evaluated by TUNEL and flow cytometry （Annexin V-FITC/PI assay）. Results 8 and 12 hrs after AOP induction, serum amylase levels were （ 1198 ± 687） U/L and （ 1698 ± 1103 ） U/L respectively; serum insulin levels were （ 8.1 ± 5.8 ） ng/ml and （ 12.7 ± 6.9 ） ng/ml respectively ; serum glucagon levels were （ 6.8 ± 4.6 ） ng/ml and （ 7.3 ± 2.9 ） ng/ml respectively ; all these parameters were significantly high than （404 ± 222 ） U/L, （5.6 ± 2.7 ） ng/ml and （ 2.6 ± 2.1 ） ng/ml in the sham operation group （ P 〈 0.05 ）. AnnexinV FITC/PI assay confirmed apoptosis occurred both in exocrine acinus cells and endocrine pancreas islet ; and the apoptotic rate was （ 20.5 ± 11.2 ） % and （ 15.5 ± 8.9） % at 8 and 12 hrs after AOP induction, which was significantly high than （4.2 ± 1.6） % in the sham operation group （ P 〈 0.05 ）. Conclusions Cell apoptosis occurred in both acinar and islet in the model of AOP, and this may be the pathophysiological basis of endocrine hormones alterations in the model of AOP.