上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (5): 543-.doi: 10.3969/j.issn.1674-8115.2012.05.003

• 论著（基础研究） • 上一篇    下一篇

急性肝衰竭早期尾加压素Ⅱ表达及其与前炎症细胞因子TNF-α和IL-1β的关系

赵 亮, 梁冬雨, 刘亮明, 于芳萍, 叶长根   

  1. 南京医科大学附属上海松江中心医院 |上海交通大学附属第一人民医院松江分院肝病科, 上海 201600
  • 出版日期:2012-05-28 发布日期:2012-06-01
  • 通讯作者: 刘亮明, 电子信箱: liuliangming@hotmail.com。
  • 作者简介:赵 亮（1981—）, 女, 主治医师, 硕士生；电子信箱: zl1519@126.com。
  • 基金资助:

    国家自然科学基金（81070357, 30660066）

Expression of urotensin Ⅱ and its relationship with pro-inflammatory cytokines of TNF-alpha and IL-1beta in early stage of acute liver failure

ZHAO Liang, LIANG Dong-yu, LIU Liang-ming, YU Fang-ping, YE Chang-gen   

  1. Department of Hepatology, Shanghai Songjiang District Central Hospital Affiliated to Nanjing Medical University, Songjiang Branch of the First People's Hospital, Shanghai Jiaotong University, Shanghai 201600, China
  • Online:2012-05-28 Published:2012-06-01
  • Supported by:

    National Natural Science Foundation of China, 81070357, 30660066

PDF (PC)

1343

摘要：

目的 探讨小鼠急性肝衰竭（ALF）早期尾加压素Ⅱ（UⅡ）表达和分泌的动态变化及其对前炎症细胞因子的影响。方法 选择6周龄雄性BALB/c小鼠60只，随机分为模型组和预处理组，每组30只。两组均建立ALF动物模型，即采用脂多糖（LPS）50 μg/kg联合右旋半乳糖胺（D-GalN）800 mg/kg（LPS/DGalN）腹腔注射；预处理组在LPS/D-GalN注射前0.5 h，以UⅡ受体拮抗剂urantide 0.6 mg/kg尾静脉注射。两组均在LPS/D-GalN注射后0、0.5、1、2和6 h处死动物（每一时间点各6只小鼠），其中0 h作为对照（仅腹腔内注射0.2 mL无菌生理盐水）；采集动物血清和肝组织标本。采用RT-PCR及ELISA方法分别检测UⅡ、肿瘤坏死因子α(TNF-α)及白介素1β(IL-1β)mRNA和蛋白质的表达。结果 LPS/D-GalN注射后0.5 h，肝组织和血清中UⅡ的表达和分泌即快速上升并达峰值，且峰值水平持续至LPS/D-GalN攻击2 h后，至6 h时UⅡ的表达与分泌虽有所降低，但仍显著高于正常水平（P<0.05）；而TNF-α水平在LPS/D-GalN攻击后0.5 h内无明显升高（P＞0.05），直到1 h后才快速上升达峰值（P<0.05），至6 h时开始下降（P<0.05）；IL-1β的表达与分泌则直到6 h后才有明显上升（P<0.05）。Urantide的应用不仅显著抑制了LPS/D-GalN攻击诱导的UⅡ高表达，也使上调的TNF-α和IL-1β表达和分泌明显受抑（P<0.05）。结论 UⅡ可刺激TNF-α的表达，有可能作为炎症级联效应的触发者在ALF的发生或启动中发挥关键性影响。

关键词: 急性肝衰竭, 尾加压素Ⅱ, 肿瘤坏死因子α, 白介素1β, 小鼠

Abstract:

Objective To investigate the alteration of expression and secretion of urotensinⅡ(UⅡ) and its effect on pro-inflammatory cytokines in early stage of acute liver failure (ALF) in mice. Methods Sixty male BALB/c mice aged 6 weeks were randomly divided into model group and pre-treatment group, with 30 mice in each group. ALF animal models were established in model group by challenging with 50 μg/kg lipopolysaccharide (LPS) and 800 mg/kg D-galactosamine (D-GalN)(LPS/D-GalN) via intraperitoneal injection, and mice in pre-treatment group were administered with 0.6 mg/kg urantide (UⅡ receptor antagonist) via caudal vein injection 0.5 h before injection of LPS/D-GalN. Mice in both groups were sacrificed 0 h, 0.5 h, 1 h, 2 h and 6 h (6 mice at each time point in each group) after injection of LPS/D-GalN, with those sacrificed 0 h after injection as controls (intraperitoneal injection of 0.2 mL normal saline), and the samples of serum and liver tissues were taken. The expression of UⅡ, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) mRNA and protein was detected by RT-PCR and ELISA respectively.ResultsThe expression and secretion of UⅡ in serum and liver tissues rapidly increased and reached the peak 0.5 h after injection of LPS/D-GalN, lasted till 2 h after injection, decreased 6 h after injection, while still higher than the normal levels 6 h after injection (P<0.05). The level of TNF-α did not significantly increase 0.5 h after injection of LPS/DGalN (P>0.05), reached the peak 1 h after injection (P<0.05), and began to decrease 6 h after injection (P<0.05). The expression and secretion of IL1β did not significantly increase until 6 h after injection of LPS/D-GalN (P<0.05). The application of urantide significantly inhibited the increased expression of UⅡ and expression and secretion of TNF-α and IL-1β induced by LPS/D-GalN challenge (P<0.05). Conclusion UⅡ can stimulate the expression of TNF-α, and may play a key role in the pathogenesis and priming of ALF as a trigger of inflammatory cascade.

Key words: acute hepatic failure, urotensinⅡ, tumor necrosis factor-α, interleukin-1β, mouse