目的 研究CCl₄致大鼠慢性肝損傷模型的最佳造模劑量和時間，并進行代謝組學(xué)評價。方法 SPF級雄性SD大鼠隨機分為6組，即對照組和10%、20%、30%、40%、50% CCl₄組。CCl₄組sc給予不同濃度的CCl₄大豆油溶液，注射體積為2 mL/kg，每周2次，連續(xù)8周，對照組給予等體積溶劑油。大鼠于造模0、2、4、6、8周末置代謝籠12 h收集尿液，取出后采用眼球后靜脈叢穿刺法取靜脈血0.3 mL，常規(guī)分離血清；4、6周末各處死3只大鼠，取肝臟；8周末造模后處死大鼠，分離各臟器，稱質(zhì)量計算臟器系數(shù)。觀察動物體質(zhì)量、肝臟外觀和病理切片、全自動生化儀檢測血清丙氨酸轉(zhuǎn)氨酶（ALT）和天門冬氨酸轉(zhuǎn)氨酶（AST）。對10% CCl₄組尿液進行氫核磁共振（¹H-NMR）代謝組學(xué)測定，運用SPSS、MatLab7.5、SIMCA-P軟件對數(shù)據(jù)進行分析。結(jié)果 對照組大鼠體質(zhì)量保持增長；隨著CCl₄劑量增高，各模型組大鼠體質(zhì)量增長速度漸趨緩慢。造模8周后，與對照組比較，10% CCl₄組肝臟系數(shù)顯著升高（P<0.01），其他組肝臟系數(shù)無顯著性變化；20%~50% CCl₄組的脾臟、腎臟系數(shù)，20%、40%、50% CCl₄組的肺臟系數(shù)均顯著升高（P<0.05、0.01）。造模4周后，10% CCl₄組大鼠肝臟出現(xiàn)了慢性炎癥表現(xiàn)，而20%~50% CCl₄組大鼠肝臟損傷明顯，出現(xiàn)明顯壞死現(xiàn)象；6周后，造模組大鼠肝臟均出現(xiàn)肝硬化病變。造模組大鼠血清ALT和AST與對照組比較均顯著升高（P<0.05、0.01），且均出現(xiàn)了先升高后降低趨勢。代謝組學(xué)研究結(jié)果顯示，造模4周起大鼠尿液與0周可以顯著分離。共尋找到差異代謝物15個，6條重要的代謝通路，涉及能量代謝、脂質(zhì)代謝、氨基酸和核苷酸代謝等途徑。結(jié)論 慢性肝損傷模型造模劑量以10% CCl₄、2 mL/kg為宜，造模時間以4~6周為宜，代謝組學(xué)研究可以動態(tài)追蹤造模過程。

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.

國家自然科學(xué)基金項目（31770362）；山西省科技創(chuàng)新重點團隊（201605D131045-18）；地產(chǎn)中藥功效物質(zhì)研究與利用山西省重點實驗室（201605D111004）