两种失眠症小鼠模型胆汁酸组成和含量差异比较

Comparative study on bile acid composition and content difference between two kinds of insomnia mouse models

目的:探究不同复制方法的失眠症模型间胆汁酸代谢的差异。方法:采用腹腔注射对氯苯丙氨酸(PCPA)法和睡眠剥夺箱(SD)法复制了两种常用小鼠失眠症模型。在对小鼠肝功能和病理学考察的基础上,采用超高效液相色谱-串联质谱(UPLC-MS/MS)定量分析了失眠症小鼠血清、肝脏和回肠中23种胆汁酸水平的变化,采用主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)确认对变化贡献率较大的成分。结果:与正常组比较,两种失眠症模型小鼠肝脏指数显著减小(P<0.05,P<0.01),血清总胆汁酸(TBA)显著增加(P<0.01,P<0.05),肝脏和回肠存在炎性损伤。PCPA和SD两种模型比较,SD组肝脏指数和TBA变化较大,但差异无统计学意义。失眠症模型小鼠血清和回肠中的胆汁酸水平整体增高,且SD组较PCPA组明显;两组肝脏胆汁酸变化不大,略有降低的趋势。PLS-DA可以将2种模型与正常组显著区分,血清中筛选出了牛磺α-鼠胆酸(T-α-MCA)、β-鼠胆酸(β-MCA)、牛磺猪去氧胆酸钠(THDCA)、石胆酸(LCA)、牛磺脱氧胆酸钠(TDCA)和α-鼠胆酸(α-MCA)6种贡献率较大的成分。结论:失眠症模型小鼠胆汁酸循环存在紊乱现象,且SD造模胆汁酸变化更为明显,血清中胆汁酸可以作为失眠症模型肝功能异常的敏感指标。

Objective:To explore the differences in bile acid metabolism among insomnia models with different replication methods.Methods:Two commonly used mouse models of insomnia were established by intraperitoneal injection of p-chlorophenylalanine(PCPA)and sleep deprivation box(SD).The changes of 23 bile acids in serum,liver and ileum of mice with insomnia were quantitatively analyzed by UPLC-MS/MS on the basis of liver function and pathological examination.PCA and PLS-DA analysis were used to identify the components contributing to the changes.Results:Compared with normal group,the liver index of the two insomnia model mice were significantly reduced(P<0.05,P<0.01),the serum total bile acids(TBA)was significantly increased(P<0.01,P<0.05),and there was inflammatory damage in the liver and ileum.Compared with the two models of PCPA and SD,the changes of liver index and TBA in SD group were significant,but there was no statistical difference.The levels of bile acid in serum and ileum of insomnia model mice were increased,and SD group was more obvious than PCPA group.There was little change in liver bile acid between the two groups,but the tendency was slightly decreased.PLS-DA could be used to distinguish the two models from the normal group.Six components contributing significantly to the abnormal liver function were screened out in serum,including T-α-MCA,β-MCA,THDCA,LCA,TDCA andα-MCA.Conclusion:There is disturbance in bile acid circulation in insomnia model mice,and the changes of bile acid in SD group are more obvious.Bile acid in serum could be used as a sensitive indicator of abnormal liver function in insomnia model.