Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro
ABSTRACT: Rumen-protected choline (RPC) supplementation in the periparturient period has in some instances prevented and alleviated fatty liver disease in dairy cows. Mechanistically, however, it is unclear how choline prevents the accumulation of lipid droplets (LD) in liver cells. In this study, p...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | Journal of Dairy Science |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022030223005994 |
| _version_ | 1797390537630679040 |
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| author | Wenyan Lu Jingna Yang Mingyue Hu Kai Zhong Yueying Wang Yanbin Yang Juan J. Loor Guoyu Yang Liqiang Han |
| author_facet | Wenyan Lu Jingna Yang Mingyue Hu Kai Zhong Yueying Wang Yanbin Yang Juan J. Loor Guoyu Yang Liqiang Han |
| author_sort | Wenyan Lu |
| collection | DOAJ |
| description | ABSTRACT: Rumen-protected choline (RPC) supplementation in the periparturient period has in some instances prevented and alleviated fatty liver disease in dairy cows. Mechanistically, however, it is unclear how choline prevents the accumulation of lipid droplets (LD) in liver cells. In this study, primary liver cells isolated from liver tissue obtained via puncture biopsy from 3 nonpregnant mid-lactation multiparous Holstein cows (∼160 d postpartum) were used. Analyses of LD via oil red O staining, protein abundance via Western blotting, and phospholipid content and composition measured by thin-layer chromatography and HPLC/mass spectrometry were performed in liver cells cultured in choline-deficient medium containing 150 μmol/L linoleic acid for 24 h. In a subsequent experiment, lipophagy was assessed in liver cells cultured with 30, 60, or 90 µmol/L choline-chloride. All data were analyzed statistically using SPSS 20.0 via t-tests or one-way ANOVA. Compared with liver cells cultured in Dulbecco's Modified Eagle Medium alone, choline deficiency increased the average diameter of LD (1.59 vs. 2.10 µm), decreased the proportion of small LD (<2 µm) from 75.3% to 56.6%, and increased the proportion of large LD (>4 µm) from 5.6% to 15.0%. In addition, the speed of LD fusion was enhanced by the absence of choline. Among phospholipid species, the phosphatidylcholine (PC) content of liver cells decreased by 34.5%. Seventeen species of PC (PC [18:2_22:6], PC [15:0_16:1], PC [14:0_20:4], and so on) and 6 species of lysophosphatidylcholine (LPC; LPC [15:0/0:0]), PC (22:2/0:0), LPC (20:2/0:0), and so on] were decreased, while PC (14:1_16:1) and LPC (0:0/20:1) were increased. Choline deficiency increased the triglyceride (TAG) content (0.57 vs. 0.39 μmol/mg) in liver cells and increased the protein abundance of sterol regulatory element binding protein 1, sterol regulatory element binding protein cleavage activation protein, and fatty acid synthase by 23.5%, 17%, and 36.1%, respectively. Upon re-supplementation with choline, the phenotype of LD (TAG content, size, proportion, and phospholipid profile) was reversed, and the ratio of autophagy marker LC3II/LC3I protein was significantly upregulated in a dose-dependent manner. Overall, at least in vitro in mid-lactation cows, these data demonstrated that PC synthesis is necessary for normal LD formation, and both rely on choline availability. According to the limitation of the source of liver cells used, further work should be conducted to ascertain that these effects are applicable to liver cells from postpartum cows, the physiological stage where the use of RPC has been implemented for the prevention and treatment of fatty liver. |
| first_indexed | 2024-03-08T23:13:16Z |
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| institution | Directory Open Access Journal |
| issn | 0022-0302 |
| language | English |
| last_indexed | 2024-03-08T23:13:16Z |
| publishDate | 2023-12-01 |
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| record_format | Article |
| series | Journal of Dairy Science |
| spelling | doaj.art-32f13e1d58e743419187975b6805e2332023-12-15T07:22:09ZengElsevierJournal of Dairy Science0022-03022023-12-011061298689878Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitroWenyan Lu0Jingna Yang1Mingyue Hu2Kai Zhong3Yueying Wang4Yanbin Yang5Juan J. Loor6Guoyu Yang7Liqiang Han8Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. ChinaKey Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. ChinaKey Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. ChinaKey Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. ChinaKey Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. ChinaKey Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. ChinaDepartment of Animal Science and Division of Nutritional Sciences, University of Illinois, Urbana, IL 61801Key Laboratory of Animal Growth and Development of Henan Province, College of Veterinary Medicine, Henan University of Animal Husbandry and Economy, Zhengzhou 450046, P.R. ChinaKey Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. China; Corresponding authorABSTRACT: Rumen-protected choline (RPC) supplementation in the periparturient period has in some instances prevented and alleviated fatty liver disease in dairy cows. Mechanistically, however, it is unclear how choline prevents the accumulation of lipid droplets (LD) in liver cells. In this study, primary liver cells isolated from liver tissue obtained via puncture biopsy from 3 nonpregnant mid-lactation multiparous Holstein cows (∼160 d postpartum) were used. Analyses of LD via oil red O staining, protein abundance via Western blotting, and phospholipid content and composition measured by thin-layer chromatography and HPLC/mass spectrometry were performed in liver cells cultured in choline-deficient medium containing 150 μmol/L linoleic acid for 24 h. In a subsequent experiment, lipophagy was assessed in liver cells cultured with 30, 60, or 90 µmol/L choline-chloride. All data were analyzed statistically using SPSS 20.0 via t-tests or one-way ANOVA. Compared with liver cells cultured in Dulbecco's Modified Eagle Medium alone, choline deficiency increased the average diameter of LD (1.59 vs. 2.10 µm), decreased the proportion of small LD (<2 µm) from 75.3% to 56.6%, and increased the proportion of large LD (>4 µm) from 5.6% to 15.0%. In addition, the speed of LD fusion was enhanced by the absence of choline. Among phospholipid species, the phosphatidylcholine (PC) content of liver cells decreased by 34.5%. Seventeen species of PC (PC [18:2_22:6], PC [15:0_16:1], PC [14:0_20:4], and so on) and 6 species of lysophosphatidylcholine (LPC; LPC [15:0/0:0]), PC (22:2/0:0), LPC (20:2/0:0), and so on] were decreased, while PC (14:1_16:1) and LPC (0:0/20:1) were increased. Choline deficiency increased the triglyceride (TAG) content (0.57 vs. 0.39 μmol/mg) in liver cells and increased the protein abundance of sterol regulatory element binding protein 1, sterol regulatory element binding protein cleavage activation protein, and fatty acid synthase by 23.5%, 17%, and 36.1%, respectively. Upon re-supplementation with choline, the phenotype of LD (TAG content, size, proportion, and phospholipid profile) was reversed, and the ratio of autophagy marker LC3II/LC3I protein was significantly upregulated in a dose-dependent manner. Overall, at least in vitro in mid-lactation cows, these data demonstrated that PC synthesis is necessary for normal LD formation, and both rely on choline availability. According to the limitation of the source of liver cells used, further work should be conducted to ascertain that these effects are applicable to liver cells from postpartum cows, the physiological stage where the use of RPC has been implemented for the prevention and treatment of fatty liver.http://www.sciencedirect.com/science/article/pii/S0022030223005994liver cellscholinefatty liverlipid dropletlipophagy |
| spellingShingle | Wenyan Lu Jingna Yang Mingyue Hu Kai Zhong Yueying Wang Yanbin Yang Juan J. Loor Guoyu Yang Liqiang Han Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro Journal of Dairy Science liver cells choline fatty liver lipid droplet lipophagy |
| title | Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro |
| title_full | Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro |
| title_fullStr | Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro |
| title_full_unstemmed | Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro |
| title_short | Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro |
| title_sort | effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro |
| topic | liver cells choline fatty liver lipid droplet lipophagy |
| url | http://www.sciencedirect.com/science/article/pii/S0022030223005994 |
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