对乙酰氨基酚诱导小鼠药物性肝损伤中PCNA的表达

Expression Analysis of PCNA During Drug-induced Liver Injury Induced by Acetaminophen in Mice

目的研究对乙酰氨基酚(Acetaminophen,AAP)诱导的小鼠药物性肝损伤过程中增殖细胞核抗原(PCNA)表达的动态变化,进一步阐明PCNA在药物性肝损伤中的重要作用。方法利用AAP溶液(550 mg/kg)腹腔注射诱导小鼠药物性肝损伤。取正常对照小鼠和注射AAP后6、24、48、72 h的小鼠肝脏制备匀浆液,利用蛋白印迹的方法检测小鼠肝损伤过程中PCNA的表达变化;同时检测各时间点小鼠血清中AST和ALT酶活变化。结果小鼠注射对乙酰氨基酚后,血清中AST和ALT酶活逐渐升高,24 h达到高峰后逐渐降低,72 h恢复到接近正常水平。PCNA在注射对乙酰氨基酚后6 h,其表达量显著下降,在24 h表达量达到最低值,随着损伤后的修复,PCNA的表达量逐渐增高,72 h逐渐恢复接近正常水平。结论 PCNA在AAP诱导的小鼠药物性肝损伤过程中表达出现显著变化,AAP诱导的药物性肝损伤过程存在一个损伤与修复逐渐变化的过程。

Objective To study the dynamic changes of PCNA during drug-induced liver injury induced by aeetaminophen in mice and further clarify the important roles of PCNA in the drug-induced liver injury. Methods 24 healthy male mice were injected intraperitoneally acetaminophen solution （diluted with saline, 550 mg/kg）. Eye blood was respectively collected at 6,24,48,72 h after aeetaminophen injection. Serum was separated,and then AST and ALT activity were measured. At the same time, the livers of mice were prepared for 10% liver homogenate. Six mice were used at each time point. At the same time, six normal mice were used as controls. Western blotting was used to detect the expression changes of PCNA during acute liver injury in mice. Results AST and ALT activity were gradually increased with time delayed and got to the peak levels at 24 h, then gradually recovered to the normal level at 72 h after aeetaminophen injection. The expression of PCNA was significantly decreased at 6 h and got to the lowest level at 24 h. Then the amount of PCNA expression gradually returned to normal level at 72 h after AAP injection. Conclusion The expression of PCNA is significantly changed during drug-induced liver injury induced by acetaminophen in mice, which indicates that the process of acute liver induced by AAP includes gradual process of injury and repair.