重组人白细胞介素-1受体拮抗剂对急性肝衰竭小鼠肝脏保护作用的评价

Evaluation of the Livers Protective Effects of Recombinant Human Interleukin-1 Receptor Antagonist in Mice with Acute Hepatic Failure

采用(+)-D-氨基半乳糖盐酸盐(D-Ga1N)结合脂多糖(LPS)诱导小鼠急性肝衰竭(AHF)来评价重组人白细胞介素-1受体拮抗剂(rhIL-1Ra)对肝脏的保护作用。本研究用SPF级雄性C57BL/6小鼠建立AHF模型。造模前1 h,小鼠腹腔注射3、9 mg/kg的rhIL-1Ra或等体积的生理盐水(NS),注射造模药物后4 h采集小鼠血液和肝组织样本。分析各组小鼠循环中的谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)水平和肝组织切片,并用荧光定量PCR方法分析肝组织中一氧化氮合成酶2(NOS2)和白细胞介素-6(IL-6)的表达水平变化。本研究还进一步通过死亡率试验考察rhIL-1Ra对AHF模型小鼠的保护作用。结果表明,9mg/kg剂量的rhIL-1Ra可以显著抑制AHF模型小鼠循环中的ALT和AST活性以及TBIL水平,减轻肝组织坏死和肝细胞变性程度,降低肝组织中NOS2和IL-6的表达。2个剂量(3和9mg/kg)的rhIL-1Ra都可以显著地改善AHF模型小鼠的生存率。结果表明,rhIL-1Ra在AHF药物开发和临床应用中具有潜在价值。

(+)-D-galactosamine hydrochloride (D-GalN) and lipopolysaccharide(LPS)-induced mouse model of acute hepatic failure (AHF) was used to investigate the livers protective effects of recombinant human interleukin-1 receptor antagonist (rhIL-1Ra). Male specific pathogen-free (SPF) C57BL/6 mice were given a single intraperitoneal injection of rhIL-1Ra (3 or 9 mg/kg) or equal volume of normal saline(NS) 1 h before D-GalN/LPS administration. Samples of blood and liver tissue were collected after 4 h of D-GalN/LPS treatment. The alanine aminotransferase (ALT) activity, aspartate transaminase (AST) activity and total bilirubin (TBIL) level in serum were analyzed. For histological analysis, liver specimen was photographed. Expression of nitricoxidesynthase 2 (NOS2) and interleukin-6 (IL-6) in the livers were detected by quantitative real time-PCR. The data showed that rhIL-1Ra could significantly suppress ALT activity, AST activity and TBIL level in AHF model mice. Histologically, rhIL-1Ra reduced hepatocytes necrosis and protected the cell shape in liver of AHF mice. Besides, NOS2 and IL-6 mRNA level in the liver were much lower in administration of rhIL-1Ra group than in AHF model group. 3 and 9 mg/kg rhIL-1Ra in two treatment groups could dramatically improve the survival rate in AHF model mice. The results showed that rhIL-1Ra has potential value in drug development and clinic application in AHF.