Abstract
Background20(S)-hydroxycholesterol (20(S)) potentially reduces adipogenesis in mammalian cells. The role of this oxysterol and molecular mechanisms underlying the adipogenesis of preadipocytes from laying hens have not been investigated. This study was conducted to 1. Analyze genes differentially expressed between preadipocytes treated with an adipogenic cocktail (DMIOA) containing 500 nM dexamethasone, 0.5 mM 3-isobutyl-1-methylxanthine, 20 μg/mL insulin and 300 μM oleic acid (OA) and control cells and 2. Analyze genes differentially expressed between preadipocytes treated with DMIOA and those treated with DMIOA + 20(S) using Affymetrix GeneChip® Chicken Genome Arrays.ResultsIn experiment one, where we compared the gene expression profile of non-treated (control) cells with those treated with DMIOA, out of 1,221 differentially expressed genes, 755 were over-expressed in control cells, and 466 were over-expressed in cells treated with DMIOA. In experiment two, where we compared the gene expression profile of DMIOA treated cells with those treated with DMIOA+20(S), out of 212 differentially expressed genes, 90 were over-expressed in cells treated with DMIOA, and 122 were over-expressed in those treated with DMIOA+20(S).Genes over-expressed in control cells compared to those treated with DMIOA include those involved in cell-to-cell signaling and interaction (IL6, CNN2, ITGB3), cellular assembly and organization (BMP6, IGF1, ACTB), and cell cycle (CD4, 9, 38). Genes over-expressed in DMIOA compared to control cells include those involved in cellular development (ADAM22, ADAMTS9, FIGF), lipid metabolism (FABP3, 4 and 5), and molecular transport (MAP3K8, PDK4, AGTR1). Genes over-expressed in cells treated with DMIOA compared with those treated with DMIOA+20(S) include those involved in lipid metabolism (ENPP2, DHCR7, DHCR24), molecular transport (FADS2, SLC6A2, CD36), and vitamin and mineral metabolism (BCMO1, AACS, AR). Genes over-expressed in cells treated with DMIOA+20(S) compared with those treated with DMIOA include those involved in cellular growth and proliferation (CD44, CDK6, IL1B), cellular development (ADORA2B, ATP6VOD2, TNFAIP3), and cell-to-cell signaling and interaction (VCAM1, SPON2, VLDLR).ConclusionWe identified important adipogenic regulators and key pathways that would help to understand the molecular mechanism of the in vitro adipogenesis in laying hens and demonstrated that 20(S) is capable of suppressing DMIOA-induced adipogenesis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1231-z) contains supplementary material, which is available to authorized users.
Highlights
Adipogenesis is the process in which preadipocytes become adipocytes, and it is one of the most intensively studied models of cellular differentiation
The effect of DMIOA on the adipogenesis and expression of key adipogenic transcripts The results of our pre-experiment investigation indicated that, preadipocytes treated with DMIOA had remarkably higher lipid accumulation (Figure 1B) and significantly higher (P < 0.05) expression of key adipogenic transcripts such as fatty acid binding protein 4 (FABP4) and The core enhancer binding protein beta (C/EBPβ) compared with non-treated cells (Figure 1C)
Genes over-expressed in control cells compared to those treated with DMIOA include those involved in cell-to-cell signaling and interaction such as interleukin 6 (IL6), calponin 2 (CNN2), and integrin beta 3 (ITGB3), cell morphology such as ATPase, Ca++ transporting, plasma membrane 2 (ATP2B2), insulin like growth factor 3 (IGFB3), and inhibin beta A (INHBA), cellular assembly and organization such as bone morphogenetic protein 6 (BMP6), insulin like growth factor 1 (IGF1), beta actin (ACTB), and adenylyl cyclase-associated Protein 2 (CAP2), cellular function and maintenance such as integrin alpha 4 (ITGA4), integrin beta 2 (ITGB2), and growth and differentiation factor 9 (GDF9), and cell cycle such as CD4, 9 and 38 molecules (CD4, CD9, CD38), and cyclin-dependent kinase inhibitor 2B (CDKN2B)
Summary
Adipogenesis is the process in which preadipocytes become adipocytes, and it is one of the most intensively studied models of cellular differentiation. The increase in adipose tissue mass results from multiplication of fat cells through a process called adipogenesis, where undifferentiated precursor cells (preadipocytes) differentiate into fat cells [2]. The critical step in these events is the activation of the transcription factor CCATT enhancer-binding protein beta (C/EBPβ) by mitogen activated protein kinase (MAPK) and glycogen synthase kinase-3 beta (GSK3β) [6]. The activated C/EBPβ triggers transcription of peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and CCATT enhancer-binding protein alpha (C/EBPα), which in turn additively activate the expression of genes responsible for the development of mature adipocytes [3]
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