PMID- 22609461
OWN - NLM
STAT- MEDLINE
DCOM- 20130805
LR  - 20130222
IS  - 1638-6183 (Electronic)
IS  - 0300-9084 (Linking)
VI  - 95
IP  - 3
DP  - 2013 Mar
TI  - Modification of the lipidome in RAW264.7 macrophage subjected to stable silencing 
      of oxysterol-binding proteins.
PG  - 538-47
LID - S0300-9084(12)00189-7 [pii]
LID - 10.1016/j.biochi.2012.05.004 [doi]
AB  - Oxysterol-binding protein homologs (ORPs) are implicated in lipid metabolism, 
      vesicle transport and cell signaling. In this study we generated RAW264.7 cells 
      with ORP1L, ORP3, or ORP8 silenced using shRNA lentiviruses. The lipidome of the 
      cells under basal serum-free culture conditions or as treated with oxidized LDL 
      (oxLDL), enzymatically modified LDL (E-LDL), or lipopolysaccharide (LPS) was 
      analyzed by mass spectrometry. Reduction in each ORP resulted in distinct and 
      complex effects on macrophage lipidome. Under basal conditions, ORP1L silencing 
      had strongest effects on phosphatidylinositols (PI, increase), free cholesterol 
      (FC, increase), and cholesteryl esters (CE, increase). ORP3 silencing affected 
      most the glucosyl ceramides (GluCer, decrease) and PE-plasmalogens (PE-pl, 
      decrease), while ORP8 silencing increased FC and CE, and decreased GluCer and 
      PE-pl. Upon LPS treatment, the ORP effects were modified: under these conditions 
      ORP1L silencing caused increase of Cer, ORP3 silencing decrease of PI, and ORP8 
      silencing decrease of PI and increase of PE, not detectable under basal 
      conditions. The lipid species data were subjected to multivariate statistical 
      analysis of principal components, revealing numerous specific alterations upon 
      ORP silencing. The cells cultured in basal conditions or treated with LPS showed 
      qualitatively different responses. However, in LPS-stimulated cells silencing of 
      any of the three ORPs decreased the relative amount of arachidonic 
      acid-containing PI species, increased the corresponding PE species, and favored 
      16-carbon sphingomyelin (SM) species at the expense of the 24-carbon ones. As a 
      conclusion, the present study reveals the distinct and sophisticated roles of 
      different ORP proteins as regulators of macrophage lipid composition, with 
      implications for inflammatory signaling.
CI  - Copyright (c) 2012 Elsevier Masson SAS. All rights reserved.
FAU - Vihervaara, Terhi
AU  - Vihervaara T
AD  - Minerva Foundation Institute for Medical Research, Biomedicum 2U, Helsinki, 
      Finland.
FAU - Kakela, Reijo
AU  - Kakela R
FAU - Liebisch, Gerhard
AU  - Liebisch G
FAU - Tarasov, Kirill
AU  - Tarasov K
FAU - Schmitz, Gerd
AU  - Schmitz G
FAU - Olkkonen, Vesa M
AU  - Olkkonen VM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120516
PL  - France
TA  - Biochimie
JT  - Biochimie
JID - 1264604
RN  - 0 (Lipopolysaccharides)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Receptors, Steroid)
RN  - 0 (oxysterol binding protein)
SB  - IM
MH  - Animals
MH  - Atherosclerosis/metabolism
MH  - Cell Line
MH  - *Gene Silencing
MH  - Lentivirus/genetics
MH  - Lipid Metabolism/drug effects/*genetics
MH  - Lipopolysaccharides/pharmacology
MH  - Macrophages/drug effects/*metabolism
MH  - Mice
MH  - RNA, Small Interfering/genetics
MH  - Receptors, Steroid/*deficiency/*genetics
EDAT- 2012/05/23 06:00
MHDA- 2013/08/06 06:00
CRDT- 2012/05/22 06:00
PHST- 2012/03/20 00:00 [received]
PHST- 2012/05/04 00:00 [accepted]
PHST- 2012/05/22 06:00 [entrez]
PHST- 2012/05/23 06:00 [pubmed]
PHST- 2013/08/06 06:00 [medline]
AID - S0300-9084(12)00189-7 [pii]
AID - 10.1016/j.biochi.2012.05.004 [doi]
PST - ppublish
SO  - Biochimie. 2013 Mar;95(3):538-47. doi: 10.1016/j.biochi.2012.05.004. Epub 2012 
      May 16.