Chylomicron production is repressed by RPTOR knockdown, R-α-lipoic acid and 4-phenylbutyric acid in human enterocyte-like Caco-2 cells.

Abstract

Although the role of mechanistic target of rapamycin complex 1 (mTORC1) in lipid metabolism has been the subject of previous research, its function in chylomicron production is not known. In this study, we created three stable human colorectal adenocarcinoma Caco-2 cell lines exhibiting normal, low, or high mTORC1 kinase activity, and used these cells to investigate the consequences of manipulating mTORC1 activity on enterocyte differentiation and chylomicron-like particle production. Constitutively active mTORC1 induced Caco-2 cell proliferation and differentiation (as judged by alkaline phosphatase activity) but weakened transepithelial electrical resistance (TEER). Repressed mTORC1 activity due to the knockdown of RPTOR significantly decreased the expression of lipogenic genes FASN, DGAT1, and DGAT2, lipoprotein assembly genes APOB and MTTP, reduced protein expression of APOB, MTTP, and FASN, downregulated the gene expression of very long-chain fatty acyl-CoA ligase (FATP2), acyl-CoA binding protein (DBI), and prechylomicron transport vesicle-associated proteins VAMP7 (vesicle-associated membrane protein 7) and SAR1B (secretion associated Ras related GTPase 1B) resulting in the repression of apoB-containing triacylglycerol-rich lipoprotein secretion. Exposure of Caco-2 cells harboring a constitutively active mTORC1 to short-chain fatty acid derivatives, R-α-lipoic acid and 4-phenylbutyric acid, downregulated chylomicron-like particle secretion by interfering with the lipidation and assembly of the particles, and concomitantly repressed mTORC1 activity with no change to Raptor abundance or PRAS40 (Thr246) phosphorylation. R-α-lipoic acid and 4-phenylbutyric acid may be useful to mitigate intestinal lipoprotein overproduction and associated postprandial inflammation.

• Authors: Bo He, Zhigang Wang, Régis Moreau
• Journal: The Journal of nutritional biochemistry (J Nutr Biochem) Vol. 108 Pg. 109087 (10 2022) ISSN: 1873-4847 [Electronic] United States
• PMID: 35691593 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S.)
• Copyright: Copyright © 2022 Elsevier Inc. All rights reserved.

Chemical References

• Apolipoproteins B
• Chylomicrons
• Phenylbutyrates
• RPTOR protein, human
• Regulatory-Associated Protein of mTOR
• Thioctic Acid
• 4-phenylbutyric acid
• Mechanistic Target of Rapamycin Complex 1
• SAR1B protein, human
• Monomeric GTP-Binding Proteins
• Coenzyme A Ligases
• long-chain-fatty-acid-CoA ligase

Topics

• Apolipoproteins B (metabolism)
• Caco-2 Cells
• Chylomicrons (metabolism)
• Coenzyme A Ligases (metabolism)
• Enterocytes (drug effects, metabolism)
• Humans
• Mechanistic Target of Rapamycin Complex 1 (metabolism)
• Monomeric GTP-Binding Proteins (genetics, metabolism)
• Phenylbutyrates (metabolism, pharmacology)
• Regulatory-Associated Protein of mTOR (genetics, metabolism)
• Thioctic Acid (metabolism, pharmacology)