Objective To investigate the effects of urantide on the signal molecules of NF-κB pathway in mice with acute liver failure. Methods Twenty four male BALB/c mices were randomly divided into 4 equal groups to be pre-treated with urantide or normal saline (NS) by tail intravenous injection. The mices were attacked with lipopolysaccharide (LPS)/ D-galactosamine (GalN) or NS via intraperitoneal injection 30 minutes later. Livers were sampled 12 h after the attack, and subjected to extract plasmosin and nucleoprotein. Protein expression was detected by Western blot; DNA binding activity of NF-κB was evaluated by EMSA. Results There were not statistical differences in IκB-α protein expression of the cytoplasm among the four groups (P>0. 05); However, the levels of p-IκB-α protein were significantly higher in model (urantide-LPS/GalN + ) and pretreatment model ( urantide + LPS/GalN + ) than those in control ( urantide- LPS/GalN-) and pretreatment control group (urantide + LPS/GalN-) [(0. 63 ± 0. 10) , (0. 31 ± 0. 05) vs (0. 14 ± 0. 02) , (0. 14 ± 0. 01) , P< 0. 01], respectively. Compared to model group, pretreatment model mice had significant lower levels of cytoplamic p-IκB-α protein [(0. 31 ± 0. 05) vs (0. 63 ± 0. 10) ,P<0. 01]. Nuclear NF-κB p65 levels were also significantly higher in model and pretreatment model group than those in control and pretreatment control group [(1. 11 ± 0. 32), (0. 69 ± 0. 14) vs (0. 10 ± 0.03), (0. 13 ±0.01), P<0. 01], respectively. While p65 levels of pretreatment model were lower than those of model group [(0. 69 ± 0. 14) vs (1.11+ 0. 32), P<0. 05]. In addition, DNA binding activities of NF-κB were significantly higher in model and pretreatment model group than those in control and pretreatment control group [(4. 68 ± 1. 03) , (1. 60 ± 0. 37) vs (1.00±0. 16), (1.01 ±0. 10), P<0.01], respectively. However, pretreatment model group had a lower DNA binding activity of NF-κB level than that in model group [(1. 60 ± 0. 37) vs (4. 68 ± 1. 03), P<0. 01]. Conclusion NF-κB signal activation induced by LPS/GalN attack can be inhibited by urantide, an antagonist of UT receptor.%目的 探讨urantide对急性肝功能衰竭(ALF)小鼠肝内NF-κB信号通路分子的影响.方法 24只雄性BALB/c小鼠被随机分为4组(6只/组),分别行urantide或0.9％氯化钠溶液尾静脉注射预处理.半小时后,以脂多糖(LPS)/D-半乳糖胺(D-GalN)或0.9％氯化钠溶液腹腔注射进行攻击.12h后采集肝组织标本,分别进行胞质蛋白和核蛋白抽提；蛋白质表达采用Western印迹方法；NF-κB与DNA的结合活性通过EMSA检测.结果 胞质IκB-α蛋白水平在4组之间的差异无统计学意义(P＞0.05),而p-IκB-α蛋白水平在模型组(urantide-,LPS/D-GalN+)和预处理模型组(urantide+,LPS/D-GalN+)较正常对照组(urantide-,LPS/D-GalN-)和预处理对照组(urantide+,LPS/D-GalN-)显著升高[(0.63±0.10)、(0.31±0.05)比(0.14±0.02)、(0.14±0.01),P＜0.01],且在预处理模型组较模型组显著降低[(0.31±0.05)比(0.63±0.10),P＜0.01]；NF-κB p65蛋白水平在模型组和预处理模型组较正常对照组和预处理对照组显著升高[(1.11±0.32)、(0.69±0.14)比(0.10±0.03)、(0.13±0.01),P＜0.01],而与模型组相比,预处理模型组p65蛋白水平降低[(0.69±0.14)比(1.11±0.32),P＜0.05]；NF-κB与DNA的结合活性在模型组和预处理模型组较正常对照和预处理对照组显著升高[(4.68±1.03)、(1.60±0.37)比(1.00±0.16)、(1.01±0.10),P＜0.01],而与模型组相比,预处理模型组NF-κB的DNA结合活性则显著降低[(1.60±0.37)比(4.68±1.03),P＜0.01].结论 UT受体拮抗剂urantide可抑制LPS/D-GalN攻击诱导的NF-κB通路分子的表达和活性.
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