细胞凋亡在重症急性胰腺炎并发急性肺损伤中的作用机制  

Action mechanism of apoptosis in the development of acute lung injury after severe acute pancreatitis

作　　者：石占利[1] 孙静[1] 李志会[1] 任丹红[1] 章佳颖[1] 方堃[1] 

机构地区：[1]浙江省中西医结合医院重症医学科,杭州310003

出　　处：《解放军医学杂志》2017年第11期974-978,共5页Medical Journal of Chinese People's Liberation Army

基　　金：浙江省自然科学基金(LY13H290008);浙江省医药卫生科技计划项目(2011KYB075)~~

摘　　要：目的探讨细胞凋亡在重症急性胰腺炎(SAP)并发急性肺损伤(ALI)中的作用,并从Notch/Hes信号转导方面探讨其作用机制。方法 50只健康雄性SD大鼠随机分为假手术组(n=8)和模型组(n=42),模型组采用胰胆管逆行注射3.5%牛磺胆酸钠的方法建立SAP并发ALI模型,并随机分为模型3h组(n=10)、模型6h组(n=10)、模型12h组(n=10)和模型24h组(n=12)。计算肺组织湿/干重比值(W/D),光镜下进行胰腺和肺组织病理评分,酶化学法测定肺组织髓过氧化物酶(MPO)活性,放射免疫分析法测定肺组织肿瘤坏死因子α(TNF-α)水平,TUNEL法检测肺组织细胞凋亡并计算凋亡指数(AI),Western blotting方法测定Notch-1、Hes-1和Hes-5蛋白的表达水平。结果模型大鼠的肺组织W/D呈现逐级增高趋势,各组均明显高于假手术组(P<0.01),其中模型12h组达到峰值;各模型组胰腺病理评分和肺组织病理评分均较假手术组明显提高(P<0.01),模型12h组最高;各模型组肺组织MPO和TNF-α水平改变与肺组织病理评分呈现相同趋势;各模型组大鼠肺组织细胞凋亡指数(AI)均较假手术组明显增高(P<0.01)。Western blotting检测结果提示,4个模型组Notch-1蛋白表达水平均较假手术组下降,其中模型3h、模型6h和模型12h三组与假手术组比较差异有统计学意义(P<0.05),模型12h组Notch-1蛋白表达水平最低,明显低于模型3h组、模型6h组和模型24h组(P<0.01)。相关性分析显示,肺组织AI与肺损伤病理评分呈显著负相关(r=–0.834,P<0.01);模型Notch-1蛋白表达水平与肺组织AI呈显著正相关(r=0.515,P=0.004),与肺组织W/D(r=–0.593,P=0.001)、肺组织病理评分(r=–0.306,P=0.002)、MPO活性(r=–0.687,P<0.01)及TNF-α水平(r=–0.574,P=0.001)呈显著负相关。结论 SAP并发ALI过程中,肺组织细胞凋亡增加对减轻肺损伤起重要作用,SAP时Notch-1信号表达受限是肺损伤恶化的重要机制。Objective To explore the role of apoptosis in the development of acute lung injury（ALI） after severe acute pancreatitis（SAP） and its mechanism via Notch/Hes signal transduction pathway in the pathologic process. Methods Fifty healthy male Sprague Dawley rats were randomly divided into, sham group（n=8） and model group（n=42）. Tissue samples of model group were collected randomly at 3（n=10）, 6（n=10）, 12（n=10） and 24 h（n=12） after model establishment. Tissue collection of Sham group was conducted at 3 h. Left lung wet/dry weight ratio（W/D） was calculated; Histological scores of pancreatic and lung tissues were assessed under microscope; myeloperoxidase（MPO） and tumor necrosis factor-α（TNF-α） of lung tissues were determined by enzymatic-chemical method and radioimmunoassay method respectively. Apoptosis of lung cells was evaluated by TUNEL assay followed by calculation of apoptosis index. Protein levels of Notch-1, Hes-1 and Hes-5 were also detected semi-quantitatively by Western blotting. Results The lung tissue W/D of model rats exhibited a gradual increment at the prior 12 hours, and the ratio was significantly higher than that of sham group at each time point（P〈0.01） while it reached the peak at 12 h time point. Pancreatic and lung pathological scores of model groups were increased at all time points and significantly higher than sham group（P〈0.01）. Lung pathological scores of model groups achieved a peak at 12 h. Meanwhile, lung MPO and TNF-α of model groups showed the same increment trend at each time points. The apoptosis index（AI） of lung cells in model groups were higher than that in sham group（P〈0.01）. Compared with sham group, the model groups showed lower protein expression levels of Notch-1, especially at 3, 6 and 12 h（P〈0.05）. Notch-1 protein expression level of 12 h group was lower than that of 3, 6 and 24 h groups（P〈0.01）. Correlation analysis found that apoptosis of lung cells was significantly negatively related to histo

关 键 词：细胞凋亡 重症急性胰腺炎 急性肺损伤 信号转导 

分 类 号：R576.1[医药卫生—消化系统]