目的 研究CCl₄致大鼠慢性肝损伤模型的最佳造模剂量和时间，并进行代谢组学评价。方法 SPF级雄性SD大鼠随机分为6组，即对照组和10%、20%、30%、40%、50% CCl₄组。CCl₄组sc给予不同浓度的CCl₄大豆油溶液，注射体积为2 mL/kg，每周2次，连续8周，对照组给予等体积溶剂油。大鼠于造模0、2、4、6、8周末置代谢笼12 h收集尿液，取出后采用眼球后静脉丛穿刺法取静脉血0.3 mL，常规分离血清；4、6周末各处死3只大鼠，取肝脏；8周末造模后处死大鼠，分离各脏器，称质量计算脏器系数。观察动物体质量、肝脏外观和病理切片、全自动生化仪检测血清丙氨酸转氨酶（ALT）和天门冬氨酸转氨酶（AST）。对10% CCl₄组尿液进行氢核磁共振（¹H-NMR）代谢组学测定，运用SPSS、MatLab7.5、SIMCA-P软件对数据进行分析。结果 对照组大鼠体质量保持增长；随着CCl₄剂量增高，各模型组大鼠体质量增长速度渐趋缓慢。造模8周后，与对照组比较，10% CCl₄组肝脏系数显著升高（P<0.01），其他组肝脏系数无显著性变化；20%~50% CCl₄组的脾脏、肾脏系数，20%、40%、50% CCl₄组的肺脏系数均显著升高（P<0.05、0.01）。造模4周后，10% CCl₄组大鼠肝脏出现了慢性炎症表现，而20%~50% CCl₄组大鼠肝脏损伤明显，出现明显坏死现象；6周后，造模组大鼠肝脏均出现肝硬化病变。造模组大鼠血清ALT和AST与对照组比较均显著升高（P<0.05、0.01），且均出现了先升高后降低趋势。代谢组学研究结果显示，造模4周起大鼠尿液与0周可以显著分离。共寻找到差异代谢物15个，6条重要的代谢通路，涉及能量代谢、脂质代谢、氨基酸和核苷酸代谢等途径。结论 慢性肝损伤模型造模剂量以10% CCl₄、2 mL/kg为宜，造模时间以4~6周为宜，代谢组学研究可以动态追踪造模过程。

Objective To explore the optimal dose and time of modeling for chronic liver injury induced by CCl₄ in rats,and to evaluate its metabolomics.Methods SPF male Sprague Dawley rats were randomly divided into six groups:control group,10%,20%,30%,40%,and 50% CCl₄ group.Rats in CCl₄ group were sc injected with CCl₄ soybean oil solution of different concentrations,the volume of injection was 2 mL/kg,twice a week for 8 weeks.Rats in control group were given the same volume of solvent oil.At the end of 0,2,4,6,and 8 weeks of modeling,rats were placed in metabolic cages to collect urine for 12 h.After removal,0.3 mL of venous blood was collected by retrobulbar venous plexus puncture,and serum was separated routinely.At the end of 4 and 6 weeks,three rats were executed to isolate the livers.At the end of 8 weeks,the organs of rats were separated,and the organ coefficient was calculated by weighting.Animal weight,organ coefficient,liver pathological changes,serum alanine aminotransferase (ALT),and aspartate aminotransferase (AST) were observed.The urine of rats in 10% CCl₄ group were determined by using ¹H-NMR metabolomics.The data were analyzed using SPSS,MatLab7.5,and SIMCA-P softwares.Results The body mass of rats in the control group kept increasing;With the increase of CCl₄ dosage,the growth rate of body mass of rats in each model group gradually slowed down.After 8 weeks,compared with the control group,the liver coefficient of 10% CCl₄ group was increased significantly (P< 0.01),while that of other groups did not change significantly;The spleen and kidney coefficients of 20%-50% CCl 4 group and lung coefficients of 20%,40% and 50% CCl₄ group were increased significantly (P< 0.05,0.01).Four weeks after modeling,rats in the 10% CCl₄ group showed chronic liver inflammation,while 20% to 50% CCl₄ group rats showed obvious liver injury and necrosis.Six weeks later,the liver of the model group rats showed cirrhosis.The ALT and AST enzymes in the serum of rats in modeling group were significantly increased compared with control group (P< 0.05,0.01),in the trend of first increased then decreased.Metabolomics studies showed that the urine of rats can be significantly separated from that of 0 week after fourth week of modeling.A total of 15 differential metabolites and six important metabolic pathways were found by urine ¹H-NMR metabolomics techniques,including energy metabolism,lipid metabolism,amino acid and nucleotide metabolism.Conclusion The optimal dose of chronic liver injury model is 0.02 mL CCl₄/100 g,and the optimal modeling time is 4-6 weeks.The metabolomics study can track the dynamic process of modeling.

国家自然科学基金项目（31770362）；山西省科技创新重点团队（201605D131045-18）；地产中药功效物质研究与利用山西省重点实验室（201605D111004）