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Effects of choline and docosahexaenoic acid on the lipidome of bovine precision-cut liver slices cultured with palmitic acid.

J. E. Rico

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06-23-2020

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Abstract:

T130
Effects of choline and docosahexaenoic acid on the lipidome of bovine precision-cut liver slices cultured with palmitic acid.
J. E. Rico*1, B. N. Tate1, W. A. Myers1, V. S�inz de la Maza-Escol�1,2, C. Chang1, L. F. Wang1,3, J. W. McFadden1. 1Cornell University Ithaca, NY, 2University of Bologna Bologna, Italy, 3Henan Agricultural University Zhengzhou, China.

Fatty acids (FA) and choline are involved in phospholipid synthesis and may influence hepatic triglyceride (TG) deposition and secretion in transition cows. Liver was biopsied from pregnant Holstein dry cows (parity = 2.4 � 0.5) and precision-cut liver slices (~200 μm) were incubated in serum-free M199 medium on titanium inserts under 95% oxygen/5% carbon dioxide within a roller culture incubator. Slices were either unsupplemented (no FA or choline), treated with 200 μM palmitic acid (PA), or treated with PA with 2 mM choline chloride (CC), 200 μM docosahexaenoic acid (DHA), or CC plus DHA for 8 h. Slice viability was assessed by measuring adenosine triphosphate (ATP) and lactate dehydrogenase leakage at h 0 and 8. Lipidomics was performed using mass spectrometry. For each experiment, 3 cows with 3 slices per cow were utilized (9 total reps/experiment). Protein-normalized, generalized log-transformed and auto-scaled data were analyzed using ANOVA. Partial least squares discriminant analysis (PLS-DA) was performed. Slice ATP and lactate dehydrogenase leakage was not different between h 0 and 8; therefore, slices were deemed viable. Lipidomics revealed 256 lipids including but not limited to TG and phosphatidylcholines (PC). The PLS-DA model distinguished slices treated with CC, DHA, or CC with DHA in the presence of PA. For component 1, changes in TG and PC were observed in response to treatment (variable importance of projection scores >1.95). Enrichment of 16:0 and 22:6 within TG was observed in slices treated with PA or DHA (e.g., TG 15:0/16:0/16:0 and TG 16:0/22:6/22:6, respectively; false discover rate [FDR] < 0.001). Treatment with PA increased PC 31:0 and 32:0; however, CC abolished these effects (FDR <0.001). Treating slices with CC or DHA increased many unsaturated PC (e.g., PC 31:1, 36:3, and 37:2; FDR <0.001), relative to PA. However, in the presence of PA, unsaturated PC were often lower with CC and DHA co-treatment, relative to DHA alone (e.g., PC 36:6; FDR <0.001). We conclude ex vivo that the ability of CC to modulate liver slice PC status depends on DHA availability.

Keywords: cow, lipidome, liver slice.