LPL-mediated lipolysis of VLDL induces an upregulation of AU-rich mRNAs and an activation of HuR in endothelial cells.
Lipoprotein lipase (LPL), the major enzyme for hydrolysis of circulating triglyceride-rich lipoproteins, is bound to the luminal surface of capillary endothelial cells. Products of LPL-mediated lipolysis, such as free fatty acids (FFA) and lipoprotein-remnants, can affect endothelial function and gene expression, and hence vascular homeostasis. In this study we tried to identify lipolysis-induced mRNAs in porcine aortic endothelial cells (ECAP) using a cDNA subtraction method. cDNA obtained from ECAP incubated with LPL and VLDL was subtracted from cDNA from cells cultured under control conditions. Analysis of the identified sequences revealed an upregulation of several mRNAs with adenine and uracil-rich elements (ARE) in their 3'-untranslated regions, such as IL-8, ESM-1 and VCAM-1. HuR, a ubiquitously expressed RNA-binding protein, is known to stabilize ARE-harboring mRNAs. Therefore, we investigated whether HuR is involved in this process and found that lipolysis induced an increased polysomal localization of HuR, which is typical for its activation pathway. In addition, the mRNAs for GM-CSF and TNF-alpha - established ARE-containing targets for HuR-mediated regulation - were upregulated by LPL-mediated lipolysis in ECAP. Differential expression of AU-rich mRNAs in response to LPL-mediated lipolysis might have an impact on physiological processes regulating lipid metabolism or pathophysiological processes promoting endothelial dysfunction and atherogenesis.
Tschernatsch
Marisa M O
MM
Institute of Molecular Biosciences, Karl-Franzens-University, Graz, Heinrichstrasse 31 a, 8010 Graz, Austria. marisa.tschernatsch@meduni-graz.at
Mlecnik
Bernhard
B
Trajanoski
Zlatko
Z
Zechner
Rudolf
R
Zimmermann
Robert
R
D000818:Animals; D000954:Antigens, Surface / Q000187:drug effects / Q000235:genetics*; D001013:Aorta, Thoracic / Q000166:cytology / Q000378:metabolism; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D015243:Cholesterol, VLDL / Q000187:drug effects / Q000378:metabolism*; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D000081035:Heterogeneous Nuclear Ribonucleoprotein D0; D034502:Heterogeneous-Nuclear Ribonucleoprotein D / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D008066:Lipolysis / Q000187:drug effects*; D008071:Lipoprotein Lipase / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins / Q000187:drug effects / Q000235:genetics*; D013552:Swine; D015854:Up-Regulation / Q000187:drug effects*
D000954:Antigens, Surface; D015243:Cholesterol, VLDL; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; C000630986:HNRNPD protein, human; D000081035:Heterogeneous Nuclear Ribonucleoprotein D0; D034502:Heterogeneous-Nuclear Ribonucleoprotein D; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D008071:Lipoprotein Lipase