Heparin inhibits NF-kappaB activation and increases cell death in cerebral endothelial cells after oxygen-glucose deprivation.
Heparin is a classic anticoagulant that is commonly used in the treatment of acute ischemic stroke (AIS). Its use remains controversial, however, due to the risk of cerebral hemorrhagic transformation. In addition to anticoagulant effects, diverse effects on transcription factors can be caused by heparin. Among the transcription factors potentially affected is nuclear factor kappa B (NF-kappaB), a protein that is reportedly related to the survival of cerebral endothelial cells. We investigated the effect of heparin on NF-kappaB activation and cell death following oxygen-glucose deprivation (OGD), an experimental model of AIS. We subjected bEnd.3 cells from a murine cerebral microvascular endothelial cell line to OGD. We examined the effect of heparin on OGD-induced NF-kappaB activation and its mechanism of action, using electrophoretic mobility shift assays, reporter gene analysis, real-time RT-PCR, Western blot analysis, and confocal microscopy. We also measured the effect of heparin on OGD-induced cell death using an MTT assay. Heparin inhibited both tumor necrosis factor alpha- and OGD-induced NF-kappaB activation. Heparin was taken up by endocytosis and then entered the nucleus. Heparin did not affect the nuclear translocation of NF-kappaB, but instead inhibited the DNA binding of NF-kappaB in the nucleus. Cells were more susceptible to OGD-induced cell death after heparin treatment. Besides producing an anticoagulation effect, heparin also inhibits NF-kappaB activation, resulting in increased susceptibility to OGD-induced cell death. This effect may be responsible for hemorrhagic transformation in patients following heparin treatment for AIS.
Lee
Jeong Ho
JH
Department of Pharmacology, Yonsei University College of Medicine, Seoul, South Korea.
Lee
Jinu
J
Seo
Gi Ho
GH
Kim
Chul Hoon
CH
Ahn
Young Soo
YS
D000818:Animals; D001932:Brain Neoplasms; D016923:Cell Death / Q000187:drug effects*; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology; D002560:Cerebrovascular Circulation / Q000187:drug effects / Q000502:physiology*; D016384:Consensus Sequence; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D017930:Genes, Reporter; D005947:Glucose / Q000172:deficiency*; D006493:Heparin / Q000494:pharmacology*; D000860:Hypoxia / Q000503:physiopathology*; D051379:Mice; D016328:NF-kappa B / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D014162:Transfection
D016328:NF-kappa B; D006493:Heparin; D005947:Glucose