Octanoylated ghrelin attenuates angiogenesis induced by oxLDL in human coronary artery endothelial cells via the GHSR1a-mediated NF-κB pathway.
OBJECTIVE
Low concentrations of oxidized low-density lipoprotein (oxLDL) promote the in vitro angiogenesis of endothelial cells and play an important role in plaque angiogenesis, which may cause plaque vulnerability and enhance the risk of intravascular thrombosis. The aim of this research was to investigate the effects of octanoylated ghrelin on oxLDL-induced angiogenesis and the underlying molecular mechanisms involved in this process.


METHODS
Human coronary artery endothelial cells (HCAECs) were incubated with 5 μg/ml oxLDL and treated with various concentrations of octanoylated ghrelin (10(-9)-10(-6)M) with or without inhibitors for 24h. Cell proliferation, migration, and in vitro angiogenesis were analyzed by bromodeoxyuridine (BrdU) staining and BrdU enzyme-linked immunosorbent assay (ELISA), transwell assay, and tube formation on Matrigel, respectively. NF-κB (nuclear factor κB) expression was determined by Western-blot analysis.


RESULTS
Treatment with oxLDL at 5 μg/ml enhanced the proliferation, migration and tube formation of HCAECs. In contrast, pretreatment with octanoylated ghrelin significantly attenuated in vitro angiogenesis in oxLDL-induced HCAECs. In addition, Western blot analysis indicated that NF-κB expression was increased after oxLDL treatment, and that this effect was significantly reversed by pretreatment with octanoylated ghrelin. However, the NF-κB inhibitor PDTC or the GHSR1a inhibitor [D-Lys3]-GHRP-6 abolished the effects of octanoylated ghrelin on the inhibition of angiogenesis and NF-κB p65 expression induced by oxLDL.


CONCLUSIONS
These findings suggest that octanoylated ghrelin attenuates angiogenesis induced by oxLDL in HCAECs via the inhibition of GHSR1a-mediated NF-κB pathway. Furthermore, octanoylated ghrelin may promote the stability of vulnerable plaques by inhibiting plaque angiogenesis.
Department of Geriatrics, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Li
L
Wang
Department of Geriatrics, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Guiqiong
G
Li
Department of Geriatrics, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China. Electronic address: chenqwcq@163.com.
Qingwei
Q
Chen
Department of Geriatrics, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Dazhi
D
Ke
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D002210:Caprylates / Q000737:chemistry; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D054439:Ghrelin / Q000031:analogs & derivatives / Q000494:pharmacology*; D006801:Humans; D008077:Lipoproteins, LDL / Q000009:adverse effects / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism; D009389:Neovascularization, Pathologic / Q000139:chemically induced / Q000517:prevention & control*; D054440:Receptors, Ghrelin / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
D020533:Angiogenesis Inhibitors; D002210:Caprylates; D054439:Ghrelin; D008077:Lipoproteins, LDL; D016328:NF-kappa B; D054440:Receptors, Ghrelin; C099252:oxidized low density lipoprotein; C031492:octanoic acid
GHSR; angiogenesis; coronary artery endothelial cell; octanoylated ghrelin; oxLDL