PARP-1通过NF-κB信号通路对重症急性胰腺炎致肠黏膜屏障损伤的作用机制  

The mechanism of poly(ADP-ribose)polymerase on the intestinal mucosal barrier injury in rat model with severe acute pancreatitis through NF-κB signaling pathway

作　　者：弥亮钰 吴自谦 潘新亭[1] 万有栋 吕韶燕 朱青云[2] 宋婧宇 王芸芸[1] 林天娇 Mi Liangyu;Wu Ziqian;Pan Xinting;Wan Youdong;Lyu Shaoyan;Zhu Qingyun;Song Jingyu;Wang Yunyun;Lin Tianjiao(Emergency Intensive Care Unit,the Affiliated Hospital of Qingdao University,Qingdao 266000,China;Department of Ultrasound,the Affiliated Hospital of Qingdao University,Qingdao 266000,China)

机构地区：[1]山东省青岛大学附属医院急诊重症医学科,266000 [2]山东省青岛大学附属医院超声科,266000

出　　处：《中华急诊医学杂志》2020年第5期675-681,共7页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金(818704440)。

摘　　要：目的探讨PARP-1[poly(ADP-ribose)polymerase-1,聚腺苷酸二磷酸核糖转移酶-1]在重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠肠黏膜屏障损伤中的作用机制。方法20只Wistar大鼠按随机数字表法分为CON组(对照组)、SAP组、3-AB组(即PARP-1抑制剂组)、3-AB-CON组,每组各5只。腹腔注射雨蛙素及脂多糖制作SAP大鼠模型,CON组仅注射等体积生理盐水,3-AB组于建模前0.5 h注射3-AB溶液30 mg/kg,3-AB-CON组腹腔注射3-AB溶液30 mg/kg 0.5 h后处理同CON组。各组大鼠建模后12 h处死,观察各组腹腔内大体改变,取心脏血、胰腺和肠组织,光镜下观察胰腺组织及肠组织病理形态学改变,采用酶联免疫吸附试验(ELISA)检测血清白细胞介素-6(IL-6)含量,使用全自动生化仪检测血清淀粉酶含量,采用蛋白免疫印迹试验(Western Blot)测定肠组织PARP-1、胞核核转录因子-κB(NF-κB)的蛋白表达,采用免疫组化试验测定肠黏膜Occludin蛋白表达。计量资料多组间比较采用单因素方差分析,方差不齐时则用秩转换的非参数检验,以P<0.05为差异有统计学意义。结果与CON组比较,SAP组大鼠腹腔明显腹水,胰腺病理明显出血坏死,肠组织绒毛结构紊乱,淀粉酶、IL-6水平升高,肠组织PARP-1、NF-κB蛋白表达明显增多,肠黏膜Occludin蛋白表达减少,说明PARP-1可诱导NF-κB活化和炎症损伤,导致胰腺及肠损伤。3-AB-CON组与CON组各指标比较差异无统计学意义。与SAP组比较,3-AB组胰腺及肠组织损伤明显减轻,淀粉酶、IL-6水平明显降低[淀粉酶(U/L)(1879.25±736.66)vs(5569.33±1993.48),IL-6(pg/mL)(77.98±20.65)vs(209.14±79.08),均P<0.05],肠组织PARP-1、胞核NF-κB蛋白表达减少[PARP-1(灰度值)(1.44±0.09)vs(1.49±0.13),NF-κB(灰度值)(0.63±0.09)vs(0.96±0.08),均P<0.05],肠黏膜Occludin蛋白表达增多[评分(6.7±1.5)vs(3.2±1.1),P<0.05]。结论抑制PARP-1表达对SAP肠黏膜屏障损伤有保护作用,其作用机制可能是通过抑制NF-�Objective To investigate the effects of poly(ADP-ribose)polymerase-1(PARP-1)in intestinal mucosal barrier injury in rat model with severe acute pancreatitis(SAP).Methods Twenty healthy male Wistar rats were divided into four groups(n=5 each group)using a random table method:control,SAP,3-aminobenzamide(3-AB),and 3-AB control groups.The SAP model was induced by intraperitoneal injection of cerulean with lipopolysaccharide.At 30 min,the rats were treated with the PARP-1 inhibitor,3-AB,or normal saline,separately.After 12 h,all rats were sacrificed to harvest pancreas tissues,intestines tissues,and blood from the hearts for index detection.Serum amylase(AMY)and interleukin(IL)-6 levels were measured using an automatic biochemical instrument and enzyme-linked immunosorbent assay(ELISA),respectively.The protein expression of PARP-1 and nuclear factor(NF-κB)were measured using Western blot and that of occludin was measured using an immunohistochemical test.One-way analysis of variance was used for comparison of multiple groups of variables.Non-parametric tests of rank conversion were used when variances were not uniform.A P<0.05 was considered statistically significant.Results Compared to the control group,the following indexes in the SAP group were significantly increased:ascites(with serious hemorrhage and necrosis in the pancreas and disordered intestinal villi),serum AMY and IL-6 levels,and the expression of PARP-1 and NF-κB.However,Occludin expression was significantly decreased.There was no significant difference between 3-AB group and 3-AB control group.Compared to the SAP group,the severity of SAP and pancreatitis-associated intestinal injury was significantly attenuated with the administration of 3-AB.Serum AMY and IL-6 levels were significantly decreased(serum AMY:1879.25±736.6 U/L vs 5569.33±1993.48 U/L;IL-6:77.98±20.65 pg/mL vs 209.14±79.08 pg/mL,both P<0.05),but the expression of PARP-1 and NF-κB were significantly increased(PARP-1:1.44±0.09 vs 1.49±0.13;NF-κB:0.63±0.09 vs 0.96±0.08,both P<0.05

关 键 词：重症急性胰腺炎 肠黏膜屏障 PARP-1 NF-ΚB OCCLUDIN蛋白 

分 类 号：R57[医药卫生—消化系统]