JNK抑制剂对仔猪肝细胞药物性损伤的缓解作用及其机理

【目的】 研究c-Jun氨基末端激酶(JNK)抑制剂SP600125对仔猪原代肝细胞药物性损伤的缓解作用及机理。【方法】 通过二步灌流法获得仔猪原代肝细胞，将肝细胞分为对照组(C)、JNK抑制剂组(SP)、模型组(M)和治疗组(T)，每组6个重复。对照组细胞不添加药物，SP组用2 μmol/L SP600125处理细胞，M组用80 μg/mL脂多糖(LPS)+20 μg/mL恩诺沙星(ENR)处理细胞，T组用80 μg/mL LPS+20 μg/mL ENR +2 μmol/L SP600125处理细胞，处理12 h后，收集上清测定谷丙转氨酶(ALT)和谷草转氨酶(AST)活性，收集细胞测定谷胱甘肽过氧化物酶(glutathione peroxidase，GSH-Px)、超氧化物歧化酶(superoxide dismutase，SOD)活性、丙二醛(malondialdehyde，MDA)含量及肝细胞核因子-1(hepatocyte nuclear factor 1，HNF-1)和谷胱甘肽-S-转移酶A1(glutathione S-transferase alpha 1，GSTA1)mRNA的相对表达量。【结果】 与对照组相比，M组肝细胞培养液上清中ALT和AST活性均显著升高(P<0.05)，M组肝细胞中GSH-Px和SOD活性、HNF-1和GSTA1 mRNA的相对表达量均极显著降低(P<0.01)，MDA含量极显著提高(P<0.01);与M组相比，T组肝细胞培养液上清中ALT、AST的活性均显著下降(P<0.05)，T组肝细胞中GSH-Px和SOD活性、HNF-1和GSTA1 mRNA的相对表达量均显著提高(P<0.05)，MDA含量显著降低(P<0.05)。【结论】 JNK抑制剂SP600125可通过调控细胞的抗氧化能力及HNF-1和GSTA1的表达缓解由LPS/ENR导致的仔猪原代肝细胞的药物性损伤。

Mitigating Effect and Mechanism of JNK Inhibitor on Drug-induced Injury of Piglet Hepatocytes

【Objective】 The aim of this study was to investigate the alleviating effect and mechanism of c-Jun N-terminal kinase (JNK) inhibitor SP600125 on the pharmacological damage of primary hepatocytes in piglets.【Method】 Piglet primary hepatocytes were obtained by two-step perfusion method,and the hepatocytes were divided into control group (C),JNK inhibitor group (SP),model group (M) and treatment group (T),with 6 replicates in each group.The cells in control group were not added with drugs,SP group cells were treated with 2 μmol/L SP600125,M group cells were treated with 80 μg/mL lipopolysaccharide (LPS) +20 μg/mL enrofloxacin (ENR),T group cells were treated with 80 μg/mL LPS+20 μg/ mL ENR+2 μmol/L SP600125.After treatment for 12 h,supernatants were collected to determine alanine transaminase (ALT) and glutamic oxalacetic transaminase (AST) activities,and cells were collected to determine the activities of glutathione peroxidase (GSH-Px),superoxide dismutase (SOD), and the content of malondialdehyde (MDA) and the relative expression of hepatocyte nuclear factor 1 (HNF-1) and glutathione S-transferase alpha 1 (GSTA1) mRNA.【Result】 Compared with control group,the activities of ALT and AST in hepatocyte culture medium were significantly increased in M group (P<0.05),the activities of GSH-Px,SOD and the relative expression of HNF-1,GSTA1 mRNA in hepatocytes were extremely significantly decreased in M group (P<0.01),and MDA content in hepatocytes was extremely significantly increased (P<0.01).Compared with M group,the activities of ALT and AST in hepatocyte culture medium were significantly decreased in T group (P<0.05),the activities of GSH-Px,SOD and the relative expression of HNF-1,GSTA1 mRNA in hepatocytes were significantly increased in T group (P<0.05),and MDA content was significantly decreased (P<0.05).【Conclusion】 JNK inhibitor SP600125 could alleviate the drug-induced injury of piglet primary hepatocytes caused by LPS/ENR by regulating the antioxidant capacity and the expression of HNF-1 and GSTA1 of cells.