高脂乳剂灌胃构建大鼠非酒精性脂肪性肝病模型可靠性的再研究

Reliability of metabolic and pathologic parameters in the high fat diet-induced nonalcoholic fatty liver disease rat model established by gastric perfusion

目的研究高脂乳剂灌胃构建大鼠非酒精性脂肪性肝病(NAFLD)模型的可靠性。方法成年Wistar雄性大鼠47只,体重(170±15)g,随机分为正常组、模型组。所有大鼠给予基础饲料,同时应用高脂乳剂灌胃法喂养模型组大鼠,而对照组大鼠用生理盐水灌胃替代。在4周、8周、12周三个时间段,光镜观察肝组织病理形态学改变;分别检测各组体重和肝脏湿重,计算肝指数(LI);检测大鼠空腹血清ALT和AST水平,观察是否存在组间差异;结果与对照组对比,模型组12周LI显著增大(P=0.000);12周时,血清ALT(P=0.030)及AST(P=0.000)较同期对照组升高;病理结果显示模型组4周肝小叶腺泡3区肝细胞少量脂肪空泡形成;模型组8周肝小叶腺泡3区肝细胞弥漫性脂肪空泡形成,少量炎性细胞浸润;模型组12周肝小叶腺泡3区弥漫性脂肪空泡形成,大量炎性细胞浸润,符合NAFLD典型的病理特征。本实验大鼠死亡率2%,模型构建成功率98%。结论脂肪乳剂灌胃12周可以成功复制大鼠NAFLD模型,该模型具有较强的临床相似性、可靠性。

Objective To evaluate the reliability of metabolic and pathologic parameters in the high fat diet-induced nonalcoholic fatty liver disease（NAFLD） rat model using a gastric perfusion method.Methods Forty-seven male Wistar rats were randomly assigned to either the normal control group（N） or the NAFLD model group（M）.There was no significant difference in body weight between the two groups.All the rats were housed under specific pathogen-free（SPF） conditions and acclimated with a normal diet for one week.Afterwards,the M group was administered a high fat diet by gastric perfusion,and the N group was administered 0.95% NS.Rats from both groups were sacrificed at the end of the 4th,8th,and 12th week for analysis.Body length and weight was recorded for each rat,as well as the liver weight for calculating the liver Index（LI）.Serum alanine aminotransferase（ALT） and aspartate aminotransferase（AST） were measured by standard biochemical methods.Formalin-fixed liver sections were stained with hematoxylin-eosin and were graded for the degree of fatty composition and inflammation.Results Two rats died during the experiment,so that only 45 were included in the analysis.The M group showed pathological changes that were typical of NAFLD,and the degree of fat degeneration was homogeneous.Compared with the N group,the M group had significantly higher LI（P=0.000）,ALT level（P=0.006）,and AST level（P=0.000）.Conclusion A NAFLD model can be successfully induced by 12 weeks of gastric perfusion with high fat diet in adult rats.The metabolic and pathologic parameters produced by this method are consistent with NAFLD.