小鼠急性酒精性肝损伤造模方法的比较

Comparison and optimization of acute alcoholic liver injury model in mice

目的模拟人类饮酒习惯,参考现有的利用酒精灌胃建立酒精性肝损伤模型的报道,探究影响模型成功的关键因素并优化小鼠酒精性肝损伤模型的构建方法。方法将150只SPF级ICR小鼠分为三个实验组讨论造模的周期和频率,造模剂的种类和用量及造模剂给予的时间间隔对实验动物的生化和生理指标所产生的影响差异。三组实验分别对小鼠进行不同剂量、不同频率、不同间隔时间的60%乙醇和53°白酒灌胃,最后计算各组小鼠的死亡率,检测血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)的活性及肝组织匀浆样本中丙二醛(MDA)、还原型谷胱甘肽(GSH)、甘油三酯(TG)的含量,并进一步对小鼠肝进行HE染色及油红染色。结果1 d造模组死亡率为20%,两次不同剂量白酒造模组死亡率为40%,4 h、6 h、8 h、12 h白酒间隔造模组死亡率依次为40%、20%、10%、0%;对血清样本中ALT和AST的含量及肝组织匀浆中MDA、GSH、TG的含量、病理学检测指标等方面进行比较发现,缩短灌胃时间间隔及增加灌胃剂量会加剧小鼠肝受损情况。结论在建立酒精性肝损伤的小鼠模型时,酒精灌胃的剂量、周期和频率会影响模型的成功率,比较而言,20 mL/kg和10 mL/kg的剂量,间隔6 h灌胃53°白酒的方法更适合用于酒精性肝损伤动物模型的构建。

Objective To investigate the factors affecting the establishment of a mouse model of alcoholic liver injury.Methods A total of 150 specific-pathogen-free male ICR mice were divided into three groups to study the possible influences of different experimental conditions,including modeling period,frequency,type,and dosage of modeling agent,and time interval between doses.Mice in the three groups received 60%ethanol and alcohol with 53%volume ratio by gavage at different doses,frequencies,and time intervals and the mortality rates were calculated.We also detected alanine aminotransferase(ALT)and aspartate transaminase(AST)serum levels and malondialdehyde(MDA),glutathione(GSH),and triglyceride(TG)levels in liver tissue homogenate.Liver pathology was examined in tissue sections.Results The mortality rates in the 1-day modeling and double-dose liquor modeling groups were 20%and 40%,respectively,and the mortality rates in the 4,6,8,and 12 h interval dosing groups were 40%,20%,10%,and 0%,respectively.Pathological indicators,including ALT,AST,MDA,GSH,TG,and tissue pathology worsened in line with shortening of the gavage interval and increased gavage dose.Conclusions The alcohol dose,length of the modeling period,and frequency of dosing affect the establishment of an alcoholic liver injury model in mice.We suggest that dosing mice with alcohol with 53%volume ratio twice a day at 20 mL/kg body weight or 10 mL/kg body weight at 6 h intervals can successfully cause liver damage.