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PMID: 20871093 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Modification in oxidative stress, inflammation, and lipoprotein assembly in response to hepatocyte nuclear factor 4alpha knockdown in intestinal epithelial cells.

The Journal of biological chemistry ·Vol. 285 ·No. 52 ·2010-12-24 ·Pages 40448-60

Marcil V, Seidman E, Sinnett D, Boudreau F, Gendron FP, Beaulieu JF, Ménard D, Precourt LP, Amre D, Levy E

Abstract

Hepatocyte nuclear factor 4α (HNF4α) is a nuclear transcription factor mainly expressed in the liver, intestine, kidney, and pancreas. Many of its hepatic and pancreatic functions have been described, but limited information is available on its role in the gastrointestinal tract. The objectives of this study were to evaluate the anti-inflammatory and antioxidant functions of HNF4α as well as its implication in intestinal lipid transport and metabolism. To this end, the HNF4A gene was knocked down by transfecting Caco-2 cells with a pGFP-V-RS lentiviral vector containing an shRNA against HNF4α. Inactivation of HNF4α in Caco-2 cells resulted in the following: (a) an increase in oxidative stress as demonstrated by the levels of malondialdehyde and conjugated dienes; (b) a reduction in secondary endogenous antioxidants (catalase, glutathione peroxidase, and heme oxygenase-1); (c) a lower protein expression of nuclear factor erythroid 2-related factor that controls the antioxidant response elements-regulated antioxidant enzymes; (d) an accentuation of cellular inflammatory activation as shown by levels of nuclear factor-κB, interleukin-6, interleukin-8, and leukotriene B4; (e) a decrease in the output of high density lipoproteins and of their anti-inflammatory and anti-oxidative components apolipoproteins (apo) A-I and A-IV; (f) a diminution in cellular lipid transport revealed by a lower cellular secretion of chylomicrons and their apoB-48 moiety; and (g) alterations in the transcription factors sterol regulatory element-binding protein 2, peroxisome proliferator-activated receptor α, and liver X receptor α and β. In conclusion, HNF4α appears to play a key role in intestinal lipid metabolism as well as intestinal anti-oxidative and anti-inflammatory defense mechanisms.

MeSH Terms
Antioxidants/metabolism Biological Transport/physiology Caco-2 Cells Epithelial Cells/cytology,metabolism Gene Knockdown Techniques Hepatocyte Nuclear Factor 4 Humans Inflammation/genetics,metabolism Interleukin-6/biosynthesis,genetics Interleukin-8/biosynthesis,genetics Intestinal Mucosa/cytology,metabolism Leukotriene B4/biosynthesis,genetics Lipid Metabolism/physiology Lipoproteins/biosynthesis,genetics Liver X Receptors Malondialdehyde/metabolism Orphan Nuclear Receptors/genetics,metabolism Oxidative Stress/physiology Oxidoreductases/genetics,metabolism PPAR alpha/genetics,metabolism Sterol Regulatory Element Binding Protein 2/genetics,metabolism
Chemicals
Antioxidants CXCL8 protein, human HNF4A protein, human Hepatocyte Nuclear Factor 4 IL6 protein, human Interleukin-6 Interleukin-8 Lipoproteins Liver X Receptors NR1H3 protein, human Orphan Nuclear Receptors PPAR alpha SREBF2 protein, human Sterol Regulatory Element Binding Protein 2 Leukotriene B4 Malondialdehyde Oxidoreductases
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Marcil Valérie
Research Institute, McGill University, Campus MGH, C10.148.6, Montreal H3G 1A4, Quebec.
Seidman Ernest
Sinnett Daniel
Boudreau François
Gendron Fernand-Pierre
Beaulieu Jean-François
Ménard Daniel
Precourt Louis-Philippe
Amre Devendra
Levy Emile
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M110.155358
Published
2010-12-24
Epub
2010-00-24
Pages
40448-60
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3003343
Subset
IM
Grants
Canadian Institutes of Health Research · CTP-82942 · Canada
External Links
PubMed source DOI: 10.1074/jbc.M110.155358 PMC Full Text View journal details
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