摘要
目的探讨高脂血症对大鼠急性胰腺炎的损伤机制。方法对高脂饮食SD大鼠和正常饮食SD大鼠均分别予雨蛙肽腹腔注射和牛磺胆酸钠胰管注射，建立4组动物模型：高脂血症急性水肿性胰腺炎组、高脂血症急性坏死性胰腺炎组、正常血脂急性水肿性胰腺炎组和正常血脂急性坏死性胰腺炎组，每组5只，采用相差双向电泳（2D-DIGE）和MALDI串联飞行时间质谱（MALDI-TOF／TOF）检测差异蛋白质，并采用免疫组化验证差异蛋白质表达。结果高脂血症急性水肿性胰腺炎和高脂血症坏死性胰腺炎动物模型胰腺组织有典型组织病理学改变。通过DeCyder图像分析软件进行匹配，高脂血症急性坏死性胰腺炎组与正常血脂急性坏死性组分析到59个差异点，39个差异点经质谱成功鉴定，23个点在高脂血症急性坏死性胰腺炎组上调，16个点下调。高脂血症急性水肿性胰腺炎组与正常血脂急性水肿性组分析到12个差异点，7个差异点经质谱成功鉴定，2个点在高脂血症急性水肿性胰腺炎组上调，5个点下调。差异蛋白质主要包括代谢酶类（脂肪酶、淀粉酶、d1抗胰蛋白酶等），内质网应激和钙代谢相关蛋白质[葡萄糖调节蛋白（GRP）78、热休克蛋白（HSP）60、蛋白二硫键异构酶等]，以及与DNA损伤修复、细胞凋亡、循环障碍、信号传导通路等相关蛋白质。淀粉酶和GRP78免疫组化验证结果与电泳结果一致。结论应用相差凝胶电泳和二级质谱的蛋白质组学手段筛选到内质网应激、细胞凋亡等相关蛋白质在高脂血症急性胰腺炎中的差异表达。高脂血症对急性胰腺炎的损伤加重通过多种途径，内质网应激相关蛋白质可能作为高脂血症相关急性重症胰腺炎的早期预警信号和临床治疗新靶点。
Objective To analyze the injury mechanism of hypeflipidemia-associated acute pancreatitis utilizing proteomics. Methods Ten SD rats were fed with high fat feed to establish hyperlipidemic models, and 10 SD rats were fed with normal feed to be used as control group. Six weeks later 5 rats from each group were intraperitoneally injected with caertdin to establish acute edematous pancreafifis models, and 5 rats from each group were injected with bile acid taurocholate into pancreatic duct to cause acute necmtizing pancreatitis. Nine hours after the establishment of models the rats were killed with their pancreases taken out. Differential protein analysis was performed using two-dimensional differential in- gel electrophoresis （2D-DIGE） and tandem mass spectrometry （MALDI-TOF/TOF） method was adopted to confirm the significantly changed proteins. Results Rat models of hypedidemia- associated acute edematous pancreatitis and rat models of hypedidemia-associated acute necrotic pancreatitis were well established. Fifty-nine spots were detected by DeCyder Analysis Software between acute necrotic pancreatitis with and without hyperlipidemia samples and 39 markedly changed protein spots were identified by MALDI-TOF-TOF. Twelve spots were detected between the samples of acute edematous pancreatitis with and without hyperlipidemia and 7 of them were identified. Differentially expressed proteins in hyperlipidemiaassociated pancreatitis included pancreatic enzymes, such as lipase, amylase, and alpha-l-antiproteinase, ER stress and calcium influx related proteins including GRP78, HSP60 and protein disulfide isomerase. Other proteins associated with DNA replication and damage repair, apoptosis, cell metabolism, circulatory dysfunction, and signal transdnction were identified in hyperlipidemia-associated acute pancreatitis. Conclusion Some proteins related to ER stress, apoptosis, circulatory dysfunction and signal transduction show differential expression in acute pancreatitis with hyperlipidemia. Hyperlipidenfia intensifies acute pancreatitis through various ways. ER stress-related proteins such as GRP78, HSP60, protein disulfide isomerase, and calreticulin can be regarded as biomarkers of early diagnosis and target of further treatment in severe acute pancreatitis.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2008年第16期1132-1136,共5页
National Medical Journal of China
基金
上海市科学技术委员会启明星科研基金资助项目（06QA14019）
关键词
胰腺炎
急性坏死性
高脂血症
电泳
凝胶
双向
内质网
Pancreatitis, acute necrotizing
Hyperlipidemias
Electrophoresis, gel, two- dimensional
Endoplasmic reticulum
