UII/UT系统对急性肝衰竭小鼠肝组织自噬水平的影响  

Effect of urotensin Ⅱ/urotensin Ⅱ receptor system on autophagy in acute liver failure in mice

作　　者：钟欢 何玉 谈志丽 刘亮明 

机构地区：[1]南京医科大学附属上海松江中心医院感染科上海交通大学附属第一人民医院松江分院感染科,上海市201600

出　　处：《世界华人消化杂志》2018年第4期228-235,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81070357;上海市松江区科学技术攻关资助项目;No.16SJGG21~~

摘　　要：目的探讨尾加压素Ⅱ(urotensin Ⅱ,UⅡ)及其受体系统(urotensin Ⅱ receptor,UT)对急性肝衰竭(acute liver failure,ALF)小鼠肝脏自噬水平的影响.方法♂Balb/c小鼠随机分为4组(每组6只).A组:健康对照组;B组:预处理对照组;C组:模型组;D组:预处理模型组.B组和D组给予0.6 mg/kg Urantide尾静脉注射预处理.30 min后,C组和D组立即以脂多糖(lipopolysaccharide,LPS)联合D-半乳糖胺(D-galactosamine,D-Gal N)腹腔注射诱导急性肝衰竭.LPS/D-Gal N攻击6 h后采集小鼠血清和肝组织样本.测量血清谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)水平以评价肝损伤情况.采用实时荧光定量PCR(realtime PCR,RT-PCR)检测Beclin-1、Atg5(autophagy related 5)、Atg7、Sqstm1(sequestosome 1)/p62和LC3 mRNA(microtubule-associated protein 1 light chain 3)水平,采用免疫印记技术(Western blot)检测LC3及p62蛋白质含量.结果模型组和预处理模型组ALT和AST水平与健康对照组和预处理对照组相比较均明显增高(P<0.01),其中预处理模型组较模型组明显降低(P<0.01).RT-PCR结果显示模型组和预处理模型组小鼠肝组织中Beclin-1、Atg5、Atg7、p62和LC3 mRNA水平均较健康对照组和预处理对照组低(P<0.05);而模型组和预处理模型组、健康对照组和预处理对照组各自间比较均无统计学差异(P>0.05).Western blot结果也显示,模型组和预处理模型组小鼠肝组织中L C3和p62蛋白水平均较健康对照组和预处理对照组低(P<0.05);而模型组和预处理模型组、健康对照组和预处理对照组各自间比较均无统计学差异(P>0.05).结论 UⅡ/UT系统对LPS/D-GalN诱导ALF小鼠下调的肝组织自噬水平无明显影响.To investigate the effect of urotensin II/urotensin II receptor （UII/UT） system on the levels of hepatic autophagy in mice with acute liver failure （ALF）.Male Balb/c mice were randomly divided into four groups （n = 6 each）： normal controls （group A）, pre- treated controls （group B）, model mice （group C）, and pre-treated model mice （group D）. Groups B and D received urantide （0.6 mg/kg body weight） via caudal vein injection. At 30 min post-injection, groups C and D were intraperitoneally injected with LPS/D-GalN to induce acute liver injury. Serum and liver tissue samples were collected 6 h later. Serum alanine transaminase （ALT） and aspartate aminotransferase （AST） levels were determined to assess liver injury. The expression of autophagy related genes such as Bedin-1, autophagy related 5 （Atg5）, Atg7, sequestosome 1 （Sqstm1/p62）, and microtubule-associated protein 1 light chain 3 （LC3） was detected by quantitative PCR. The expression of autophagic proteins LC3 and p62 was tested by Western blot. Serum ALT and AST levels in group C were significantly higher than those in groups A and B （P 〈 0.01）, while they were significantly lower in group D than in group C （P 〈 0.01）. RT-PCR analysis showed that the expression of autophagy related genes such as Beclin-1, Atg5, Atg7, p62, LC3 was downregulated in groups C and D compared to groups A and B （P 〈 0.05）, although there was no difference between groups C and D as well as between groups A and B （P 〉 0.05）. LC3II and p62 protein levels tested by Western blot were significantly lower in groups C and D than in groups A and B （P 〈 0.05）, but there was no difference between groups C and D as well as between groups A and B （P 〉 0.05）.CONCLUSION UII/UT system has no influence on the suppressed hepatic autophagy in ALF mice.

关 键 词：尾加压素Ⅱ 急性肝衰竭 自噬 URANTIDE 小鼠 

分 类 号：R575.3[医药卫生—消化系统]