一种非酒精性脂肪肝小鼠模型的建立方法

An Establishment Method of a Mouse Model of Non-alcoholic Fatty Liver Disease

[目的]建立一种由高脂高糖膳食导致小鼠患非酒精性脂肪肝病的方法,具体症状为伴有严重血脂异常、肝功能受损和炎症反应。[方法]选用SPF级雄性C57BL/6J小鼠,7~8周龄,采用随机数字法分为正常组和模型组,每组20只,模型组灌胃高脂高糖肠内营养液,正常组灌胃PBS,试验期间自由摄食基础饲料和水。于第10、12、14、16周分别处死3只小鼠,检测是否达到NAFLD脂变程度;16周后,处死全部小鼠进行常规血脂4项和肝功能检测;观察肝脏的外观,测定肝脏中TG和TC含量;另取肝脏、脾脏和腹部脂肪分别进行HE染色;测定血清中IL-6和TNF-α水平。[结果]随着灌胃时间的增加,模型组肝脏系数、TG、TC含量均呈逐渐增加的趋势,正常组对应指标变化不明显;当灌胃时间达到12周时,模型组肝脏TC含量显著高于正常组;14周时,模型组中TG、TC含量与正常组相比均显著增加;通过HE染色发现,灌胃16周时模型组肝细胞排序混乱,肝细胞脂肪严重;血液中炎症因子和肝功能水平显著高于正常组,说明灌胃高脂高糖肠内营养液16周,模型组内肝脂变严重,肝细胞损伤并出现明显的炎性病变。[结论]该研究建立NAFLD小鼠模型操作简便,检测的指标稳定性好、可重复性高且更接近模拟人类该疾病的自然发病规律,为建立深入研究NAFLD发病机制、相关防治措施和治疗药物的筛选提供了可靠的动物模型。

[Objective]A method of non-alcoholic fatty liver disease in mice caused by a high-fat,high-sugar diet was established,specifically with severe dyslipidemia,impaired liver function,and an inflammatory response.[Method]SPF male C57BL/6J mice(7-8 week-old)were selected and divided into normal and model groups by random number method,with 20 mice in each group.Mice in model group was fed with high fat and high glucose enteral nutrition,mice in the normal group was fed with PBS.Mice were freely fed with the basal diet and water during this period.Three mice were killed at 10,12,14,and 16 weeks respectively to determine whether NAFLD lipid change was achieved.All mice were killed after 16 weeks for routine lipid 4 tests and liver function tests.The appearance of the liver was observed,and the contents of TG and TC in the liver were measured.Furthermore,liver,spleen,and abdominal fat were taken for HE staining.IL-6 and TNF-αlevels in the serum were determined.[Result]The liver coefficient,TG and TC contents in the model group gradually increased with the increase of the gastric irrigation time,and the corresponding indices in the normal group did not change significantly.When the gastric irrigation time reached 12 weeks,TC content in the liver of the model group was significantly higher than that in normal group.At 14-week-age,the contents of TG and TC in the model group were significantly increased compared with the normal group.The results of HE staining revealed that the liver cells in the model group were disordered and intrahepatic cells are severely fat.The levels of inflammatory factors and liver function in the blood were significantly higher than those in the normal group.These indicated that the intragastric high-fat and high-sugar enteral nutrition solution was administered for 16 weeks.The liver lipids in the model group became severe,and the hepatocytes were damaged with obvious inflammatory lesions.[Conclusion]The establishment of NAFLD mouse model was easy to operate,and the indicators tested were stable,highly reproducible,and closer to stimulate the natural occurrence of the disease in humans.The research results provided a reliable animal model for the in-depth study of the pathogenesis of NAFLD,related prevention and treatment measures and the screening of therapeutic drugs.