C／EBPs和内质网应激相关分子在高三酰甘油血症性急性胰腺炎大鼠胰腺中的表达  

Alteration of C/EBPs and endoplasmic reticulum stress related molecules expression in pancreatic tissues in rats with hypertriglyceridemia related acute pancreatitis

作　　者：郑俊媛 吴江红[1] 陈静[1] 刘杰[1] 胡国勇[1] 王兴鹏[1] 曾悦[1] 

机构地区：[1]上海交通大学附属第一人民医院消化科,上海200080

出　　处：《中华胰腺病杂志》2016年第2期103-109,共7页Chinese Journal of Pancreatology

基　　金：基金项目：国家自然科学基金青年项目（81200322）;上海市自然科学基金面上项目（12ZR1424000）;上海市胰腺疾病重点实验室开放课题资助项目（SPDF2013004）

摘　　要：目的探讨C/EBPα、C/EBPβ和内质网应激相关分子（IRE1α和sXBP1）在高三酰甘油血症（HTG）相关性急性胰腺炎（AP）大鼠胰腺中的表达变化。方法96只SD大鼠按数字表法随机分为对照组、HTG组（高脂饮食2周）、AP组、HTG＋AP组，每组24只。采用腹腔注射雨蛙肽方法制备AP模型，造模后3、6、9、24 h分批处死大鼠。观察胰腺病理变化并予评分，检测血清TG、总胆固醇（TC）及血浆IL-1β、IL-6、TNF-α水平，实时PCR法检测胰腺组织IRE1α、sXBP1、C/EBPα、C/EBPβ mRNA表达，蛋白质印迹法检测胰腺组织IRE1α、sXBP1、NF-κB、C/EBPα和C/EBPβ蛋白表达，免疫组化法检测胰腺组织C/EBPα和C/EBPβ蛋白表达。结果高脂饮食2周后，HTG组、HTG＋AP组大鼠血清TG和TC均较对照组明显升高，差异有统计学意义（P值均〈0.05）。HTG＋AP组胰腺病理变化较AP组更严重，以9 h点表现最为显著（P〈0.05）。与AP组比较，HTG＋AP组血浆IL-1β、IL-6和TNF-α水平明显升高，9 h点差异有统计学意义（P值分别为0.011、0.034、0.027）。AP组和HTG＋AP组胰腺组织IRE1α、sXBP1、C/EBPα、C/EBPβ mRNA表达均于3 h点开始升高，分别于24、9、6、6 h点达峰值。与AP组比较，HTG＋AP组IRE1α、sXBP1、C/EBPα、C/EBPβ mRNA表达明显更高，其中IRE1α、sXBP1 mRNA 3、6、9、24 h点，C/EBPα mRNA 6、9、24 h点，C/EBPβ mRNA 6、9 h点差异均有统计学意义（P值均〈0.05）。AP组和HTG＋AP组胰腺组织IRE1α、sXBP1、NF-κB蛋白表达均于3 h点开始上调，9 h点达最高水平，HTG＋AP组较AP组上调更显著；HTG＋AP组6、9 h点C/EBPα、C/EBPβ蛋白高表达，AP组则无表达。结论C/EBPα、C/EBPβ、IRE1α、sXBP1基因参与HTG相关性AP的发病，内质网应激通路IRE1α/sXBP1和C/EBPα、C/EBPβ可能共同介导HTG性AP的胰腺病理损伤和炎症过程。Objective To investigate the alteration of C/EBP α, C/EBP β and endoplasmic reticulum stress （ERS） related molecules （IRE1 a and sXBP1 ） expression in pancreatic tissues in rats with hypertriglyceridemia related acute panereatitis. Methods Ninety six Sprague Dawley （SD） rats were divided into 4 groups ： control group, hypertriglyceridemia （HTG） group （ n = 24, fed with high fat diet for 2 weeks）, AP group （n = 24）, HTG ＋ AP group （n = 24）, and AP was induced by peritoneal injection of cerulein. The rats were sacrificed at 3, 6, 9, 24 h after AP induction, respectively. The pathological changes of the pancreatic tissues were observed and scored by HE staining. Plasma levels of IL-1 β, IL-6 and TNF-α were measured by ELISA. The expression of IRE1α, sXBP1, C/EBPα, C/EBPβ mRNA were analyzed by real time PCR. The expressions of IRE1α, sXBP1, NF-kB, C/EBPα and C/EBPβ protein were determined by WesternBlot. The expressions of C/EBPα and C/EBPβ proteins were also determined by immunohitochemistry. Results After two weeks of high fat diet, serum levels of triglyceride（TG） and total cholesterol （TC） in HTG group, HTG ＋ AP group were much higher than those of the control group, and the difference was statistically significant （ P 〈 0.05 ）. The pancreatic tissue injury was more severe in HTG ＋ AP group, particularly at 9 h （P 〈 0.05）. And plasma IL-113, IL-6 and TNF-α levels were also much higher in HTG ＋ AP group when compared with that of AP group, the differences were all significant at 9 h （P=0. 011; P =0.054; P =0. 027）. After AP induction, IRE1, sXBP1, C/EBP and C/EBPβ mRNA began to be up-regulated at 3 h, and IRE1 mRNA reached the highest level at 24 h, sXBP1 mRNA at 9 h, while C/EBP and C/EBPI3 mRNA reached the highest level at 6 h. Compared with AP group, IRE1, sXBP1, C/EBP and C/EBPβ mRNA levels were much higher in HTG ＋ AP group. In addition, as to IRE1 and sXBP1 mRNA, the difference was significant at 3, 6, 9, 24h, and C/EBP mRNA at 6,

关 键 词：胰腺炎 C/EBPΑ C/EBPΒ 内质网应激 高酯血症 大鼠 

分 类 号：R576[医药卫生—消化系统]