摘要
目的明确两种不同造模方法对酒精性肝病模型小鼠的肝脏脂变、炎症及铁沉积的影响。方法分别采用酒精液体饮食喂养C57BL/6小鼠10 d和8周,分为10 d对照组(对照饮食)、10 d模型组(酒精液体饮食)、8周对照组(对照饮食)、8周模型组(酒精液体饮食),每组10只。造模结束后留取各组小鼠血清及肝脏组织检测。采用苏木精-伊红、油红O、普鲁士蓝染色分别观察小鼠肝脏炎症、脂变、铁沉积情况。依据相应试剂盒说明书检测血清中肝功能生化指标[天冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)]、脂质相关生化指标[血清甘油三脂和总胆固醇]、铁代谢相关生化指标[血清铁调素、血清铁及总铁结合力(TIBC)、转铁蛋白饱和度(TS)]。采用Real-time PCR(qPCR)检测肝脏炎症相关基因IL-1β、IL-10以及铁代谢相关基因HAMP的表达。结果苏木精-伊红染色显示10 d和8周模型组仅少量炎症细胞浸润;油红O染色显示10 d及8周模型组较对照组均出现明显脂变;普鲁士蓝染色显示10 d及8周模型组较对照组均无明显铁沉积。10 d模型组的血清ALT、AST水平以及IL-1β表达均较10 d对照组升高,差异均有统计学意义(P<0.05),两组间IL-10表达差异无统计学意义(P>0.05)。8周模型组血清ALT、AST水平以及IL-1β、IL-10表达与8周对照组比较,差异均无统计学意义(P>0.05)。10 d模型组甘油三酯、总胆固醇水平均较10 d对照组升高,差异均有统计学意义(P<0.05);8周模型组甘油三酯、总胆固醇与对照组比较,差异均无统计学意义(P>0.05)。两组模型中血清铁均较对照组均有一定上升趋势,但差异均无统计学意义(P>0.05)。10 d模型组与10 d对照组TS水平比较,差异无统计学意义(P>0.05);8周模型组TS水平较8周对照组升高,差异有统计学意义(P<0.05)。10 d及8周模型组的TIBC水平与对照组比较,差异均无统计学意义(P>0.05)。10 d模型组与10 d对照组血清铁调素水平比较,差异无统计学意义(P>0.05);8周模型组的血清铁调素水平较8周对照组降低,差异有统计学意义(P<0.05)。肝脏HAMP基因表达在10 d和8周模型组均较对照组降低,差异均有统计学意义(P<0.05)。结论10 d及8周酒精喂养均能建立酒精性脂肪肝模型,均能引起肝脏轻度炎症,但肝脏均无明显铁沉积,可能需要延长造模时间或额外铁负荷才能建立酒精肝铁过载模型。
Objective To investigate the effects of two different modeling methods on liver inflammation,steatosis,and iron deposition in alcoholic liver disease mice.Methods C57BL/6 mice were fed with alcohol liquid diet for 10 days and 8 weeks respectively,and were divided into 10 day control group(control diet),10 day model group(alcoholic liquid diet),8 week control group(control diet),8 week model group(alcoholic liquid diet),with 10 mice in each group.Serum and liver tissues were harvested for further study.The level of mice hepatic inflammation,steatosis,and iron deposition were evaluated by hematoxylin-eosin staining,oil red O staining,and prussian blue staining.The corresponding test kits were used to detect the level of the liver functional markers[aspartate aminotransferase(AST)and alanine aminotransferase(ALT)]and the lipid metabolism associated biomarkers[triacylglycerol,total cholesterol]and the iron metabolism associated biomarkers[hepcidin,iron and total iron binding capacity(TIBC),transferrin saturation(TS)]in serum.Real-time PCR(qPCR)was used to detect the expression of inflammation and iron metabolism-related genes:IL-1β,IL-10 and HAMP.Results According to the hematoxylin-eosin staining,no apparent inflammatory cell infiltration was found in livers from 10 days and 8 weeks ethanol model compared to those from the control group.Oil red O staining showed that hepatic steatosis occurred in both 10 days and 8 weeks model group compared to those of the control groups.Meanwhile,no conspicuous liver iron deposition could be observed in livers from10 days and 8 weeks model group mice.The levels of serum ALT,AST and IL-1βin the 10 days model group were significantly higher than those in the 10 days control group,the differences were statistically significant(P<0.05),there was no significant difference in the expression of IL-10 between the two groups(P>0.05).There was no significant difference in serum ALT,AST,IL-1β,IL-10 expression between the 8 weeks model group and the 8 weeks control group(P>0.05).The levels of triglyceride and total cholesterol in the 10 days model group were significantly higher than those in the 10 days control group,the differences were statistically significant(P<0.05),there was no significant difference in triglyceride and total cholesterol between the 8 weeks model group and the 8 weeks control group(P>0.05).The serum iron in the two model groups had a certain increase trend compared with the control group,but the difference was not statistically significant(P>0.05).There was no significant difference in TS level between the 10 days model group and the 10 days control group(P>0.05);the level of TS in the 8 weeks model group was significantly higher than that in the 8 weeks control group,the difference was statistically significant(P<0.05).There was no significant difference in the TIBC level between the 10 days model group and the 8 weeks model group and the control group(P>0.05).There was no significant difference in serum ferrimodulin levels between the 10 days model group and the 10 days control group(P>0.05);the level of serum ferrimodulin in the 8 weeks model group was significantly lower than that in the 8 weeks control group,the difference was statistically significant(P<0.05).The expression of HAMP gene reduced in livers from 10 days and 8 weeks model groups compare to those of the control groups,the differences were statistically significant(P<0.05).Conclusion Both 10 days and 8 weeks ethanol feeding could establish an alcoholic fatty liver mice model,accompanied by slight liver inflammation,but no liver iron deposition.Longer modeling time or additional iron stimulation may be required to develop an iron overload model of ALD.
作者
谢超
王晓凡
刘天会
王萍
范旭
刘琳
丛敏
XIE Chao;WANG Xiao-fan;LIU Tian-hui(Liver Research Center,Beijing Friendship Hospital,Capital Medical University,Beijing Key Laboratory of Translational Medicine in Liver Cirrhosis,National Clinical Research Center of Digestive Disease,Beijing 100050,China)
出处
《临床和实验医学杂志》
2023年第6期561-565,共5页
Journal of Clinical and Experimental Medicine
基金
国家自然科学基金(编号:82170614)
王宝恩肝纤维化基金(编号:CFHPC 2021042)。
关键词
酒精
小鼠模型
脂肪肝
Alcohol
Mice model
Fatty liver
