Changes of serum and liver bile acid profiles in a mouse model of metabolic associated fatty liver disease induced by a methionine-choline-deficient diet

doi:10.12307/2021.111

Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (26): 4137-4144.doi: 10.12307/2021.111

Changes of serum and liver bile acid profiles in a mouse model of metabolic associated fatty liver disease induced by a methionine-choline-deficient diet

Yang Hailin1,2,3, Zhang Dingqi1,2,3, Chen Gaofeng1,2,3, Zhang Congcong4, Chen Jiamei1,2,3, Wang Xiaoning5, Liu Wei1,2,3, Liu Ping1,2,3,5

1Institute of Liver Diseases, 2Shanghai Key Laboratory of Traditional Chinese Medicine, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 3Liver and Kidney Diseases Key Laboratory of Ministry of Education, 4Institute of Chinese Materia Medica,5Institute of Interdisciplinary Science, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

Received:2020-08-07 Revised:2020-08-11 Accepted:2020-09-17 Online:2021-09-18 Published:2021-04-26
Contact: Liu Ping, MD, Professor, Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Shanghai Key Laboratory of Traditional Chinese Medicine, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Liver and Kidney Diseases Key Laboratory of Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Institute of Interdisciplinary Science, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China Liu Wei, MD, Associate researcher, Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Shanghai Key Laboratory of Traditional Chinese Medicine, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Liver and Kidney Diseases Key Laboratory of Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China E-mail:liuliver@vip.sina.com;lwhzayl@shutcm.edu.cn
About author:Yang Hailin, Master candidate, Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Shanghai Key Laboratory of Traditional Chinese Medicine, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Liver and Kidney Diseases Key Laboratory of Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Supported by:
the National Natural Science Foundation of China, No. 81703681 (to LW) and 81530101 (to LP); Shanghai Sailing Plan for Scientific Youth Talents, No. 17YF1419800 (to LW)

Abstract

Abstract:

BACKGROUND: Methionine-choline-deficient (MCD) model is widely used to induce metabolic associated fatty liver disease. There are few dynamic changes of bile acid profiles reported in the existing relevant research; however, the regulation of bile acid is one of the important ways to intervene with the lipid metabolism of liver. Therefore, it is necessary to clarify the dynamic changes of bile acid profiles during model establishment.

OBJECTIVE: To investigate the changes of bile acid profiles in serum and liver of mice with metabolic associated fatty liver disease induced by feeding a MCD diet.
METHODS: After 1 week of adaptive feeding, 60 male C57/BL6J mice were randomly divided into 2 groups, with 30 mice in each group. One group was fed with methionine-choline-sufficient diet (MCS), and the other group was fed with MCD diet, with free access to food and water. At the end of the 4th, 6th, and 8th week, 10 mice were randomly selected from each group, with serum and liver tissue samples of mice collected. The activities of alanine aminotransferase and aspartate aminotransferase in serum and the content of triglyceride in liver tissue were detected. Hematoxylin-eosin staining, oil red O staining and sirius red staining were used for pathological evaluation. The contents of 18 kinds of bile acids in serum and liver were measured by ultra-performance liquid chromatography-tandem mass spectrometry.
RESULTS AND CONCLUSION: The test results of animal samples with different modeling time showed that, compared with the MCS diet group, the activity of alanine aminotransferase and aspartate aminotransferase in serum and the content of triglyceride in liver tissue were significantly increased in the MCD diet group at each observational time (P < 0.05). Compared with the MCS diet group, extensive hepatocyte steatosis was found in liver tissue, collagen fibers around sinusoids increased, and non-alcoholic fatty liver disease activity score increased significantly in the MCD diet group at each observational time. Compared with the MCS diet group, the content of total free bile acids and total combined bile acids and the content of bile acids except taurodeoxycholic acid in the serum of the MCD diet group showed an upward trend, and the value of cholic acid/chenodeoxycholic acid increased significantly. Compared with the MCS diet group, the contents of total free bile acids, bile acids, glycocholic acid, glycochenodeoxycholic acid, and taurocholic acid in the liver of the MCD diet group showed an increasing trend, while the contents of chenodeoxycholic acid, ursodeoxycholic acid, hyodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, taurochenodeoxycholic acid, and taurodeoxycholic acid in the liver decreased. The content of glycodeoxycholic acid decreased first and then increased with the extension of modeling time. The contents of deoxycholic acid and taurohyodeoxycholic acid remained basically unchanged, and the value of cholic acid/chenodeoxycholic acid increased significantly. All these findings show that the bile acid profiles in the serum and liver of mice with metabolic fatty liver disease induced by MCD diet change significantly, and especially the increase of cholic acid/chenodeoxycholic acid ratio may play a key role. It is suggested that the development of metabolic fatty liver disease in the MCD model may be closely related to the changes of bile acid profiles and its toxicity.

Key words: methionine-choline-deficient, diet, methionine, choline, metabolic associated fatty liver disease, bile acids

CLC Number:

Yang Hailin, Zhang Dingqi, Chen Gaofeng, Zhang Congcong, Chen Jiamei, Wang Xiaoning, Liu Wei, Liu Ping. Changes of serum and liver bile acid profiles in a mouse model of metabolic associated fatty liver disease induced by a methionine-choline-deficient diet[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(26): 4137-4144.

Figures/Tables 11

从小鼠血清胆汁酸检测结果可以看出，与蛋氨酸-胆碱充分组小鼠相比，蛋氨酸-胆碱缺乏组小鼠血清中总游离型和总结合型胆汁酸以及除牛磺脱氧胆酸外的胆汁酸，含量均呈上升趋势，胆酸/鹅去氧胆酸值显著升高；随着饲养时间增长，蛋氨酸-胆碱充分组胆汁酸含量猪去氧胆酸、甘氨胆酸、牛磺猪去氧胆酸含量显著下降；如图4所示，蛋氨酸-胆碱缺乏组与蛋氨酸-胆碱充分组的小鼠血清中胆汁酸的组成发生了较大改变，其中变化幅度较大的胆汁酸为胆酸、牛磺猪去氧胆酸、甘氨胆酸、牛磺熊去氧胆酸、牛磺脱氧胆酸等。发现造模4，6，8周时，与蛋氨酸-胆碱充分组胆酸占比(9.87%，20.16%，15.02%)相比，在蛋氨酸-胆碱缺乏组的占比(47.49%，56.74%，57.80%)分别增加了4.8，2.8，3.8倍；与蛋氨酸-胆碱充分组牛磺猪去氧胆酸占比(7.12%，4.97%，1.64%)相比，在蛋氨酸-胆碱缺乏组的占比(0.56%，0.42%，0.47%)降低了92%，92%，72%；与蛋氨酸-胆碱充分组甘氨胆酸占比(9.05%，7.18%，8.91%)相比，在蛋氨酸-胆碱缺乏组的占比(0.96%，0.79%，0.97%)降低了90%，89%，89%；与蛋氨酸-胆碱充分组牛磺熊去氧胆酸占比(7.41%，6.49%，6.76%)相比，在蛋氨酸-胆碱缺乏组的占比(2.39%，2.09%，1.37%)降低了68%，68%，80%；与蛋氨酸-胆碱充分组牛磺脱氧胆酸占比(14.73%，13.37%，12.64%)相比，在蛋氨酸-胆碱缺乏组的占比(2.40%，1.80%，1.84%)降低了84%，87%，86%。从小鼠肝脏胆汁酸检测结果可以看出，与蛋氨酸-胆碱充分组小鼠相比，蛋氨酸-胆碱缺乏组肝脏中总游离型胆汁酸以及胆汁酸胆酸、甘氨胆酸、甘氨鹅去氧胆酸、牛磺胆酸含量呈上升趋势，胆汁酸鹅去氧胆酸、熊去氧胆酸、猪去氧胆酸、牛磺熊去氧胆酸、牛磺石胆酸、牛磺鹅去氧胆酸、牛磺脱氧胆酸含量呈下降趋势，胆汁酸甘氨脱氧胆酸含量呈先下降后上升趋势，胆汁酸去氧胆酸、牛磺猪去氧胆酸含量基本不变，胆酸/鹅去氧胆酸值显著升高；随着饲养时间增长，蛋氨酸-胆碱充分组胆汁酸鹅去氧胆酸、熊去氧胆酸、去氧胆酸、猪去氧胆酸、甘氨胆酸、甘氨脱氧胆酸、牛磺脱氧胆酸含量显著下降；如图5所示，蛋氨酸-胆碱缺乏组小鼠肝脏中胆汁酸的组成发生了较大变化，其中变化显著的为胆酸、牛磺鹅去氧胆酸、牛磺猪去氧胆酸、牛磺熊去氧胆酸、牛磺脱氧胆酸。发现造模4，6，8周时，与蛋氨酸-胆碱充分组胆酸占比(3.12%，3.94%，3.47%)相比，在蛋氨酸-胆碱缺乏组的占比(18.09%，30.83%，28.91%)增加了5.8，7.8，8.3倍；与蛋氨酸-胆碱充分组牛磺鹅去氧胆酸占比(12.47%，13.20%，13.40%)相比，在蛋氨酸-胆碱缺乏组的占比(9.28%，7.82%，5.09%)降低了26%，41%，63%；与蛋氨酸-胆碱充分组牛磺猪去氧胆酸占比(6.57%，8.27%，8.84%)相比，在蛋氨酸-胆碱缺乏组的占比(4.54%，5.49%，4.35%)降低了31%，34%，51%；与蛋氨酸-胆碱充分组牛磺熊去氧胆酸占比(5.10%，5.12%，2.20%)相比，在蛋氨酸-胆碱缺乏组的占比(1.38%，1.02%，1.36%)降低了73%、81%、39%；与蛋氨酸-胆碱充分组牛磺脱氧胆酸占比(11.26%，11.17%，10.06%)相比，在蛋氨酸-胆碱缺乏组的占比(3.50%，2.46%，2.62%)降低了69%，78%，74%。此外，将血清及肝脏中胆汁酸胆酸/鹅去氧胆酸比值与肝损伤相关的生化指标(血清谷丙转氨酶、谷草转氨酶及肝组织三酰甘油)作Pearson相关分析，结果见表5，6。表中相关系数为正值说明呈正相关，表示随着肝损伤的增强胆酸/鹅去氧胆酸比值升高，相关系数的绝对值越大代表相关性越强。可以看出，4，6，8周小鼠血清中胆酸/鹅去氧胆酸比值与血清谷丙转氨酶、谷草转氨酶活性呈正相关(P < 0.05)，8周小鼠血清中胆酸/鹅去氧胆酸比值与肝组织三酰甘油含量呈正相关(P < 0.01)，且整个造模过程中的小鼠血清中胆酸/鹅去氧胆酸比值与血清谷丙转氨酶、谷草转氨酶活性及肝组织三酰甘油含量呈正相关(P < 0.01)。4，6，8周小鼠肝脏中胆酸/鹅去氧胆酸比值与血清谷丙转氨酶、谷草转氨酶活性呈正相关(P < 0.05)，6，8周小鼠肝脏中胆酸/鹅去氧胆酸比值与肝组织三酰甘油含量呈正相关(P < 0.05)，且整个造模过程中的小鼠肝脏中胆酸/鹅去氧胆酸比值与血清谷丙转氨酶、谷草转氨酶活性及肝组织三酰甘油含量呈正相关(P < 0.01)。"

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Changes of serum and liver bile acid profiles in a mouse model of metabolic associated fatty liver disease induced by a methionine-choline-deficient diet

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