RESUMEN
The response to vascular injury is a complex wound healing response involving cell proliferation, migration, remodeling and inflammation. In the present studies we employed a rat balloon angioplasty model of vascular injury to investigate the potential role of sphingolipid signaling in the response to vascular injury. The enzyme serine palmitoyltransferase (SPT) catalyzes the first committed step in de novo sphingolipid biosynthesis. We observed marked upregulation of expression of both SPT subunits in actively proliferating cells in injured vessels. This enhanced SPT expression occurs in de-differentiated fibroblasts and proliferating vascular smooth muscle cells. The upregulation is particularly apparent in the proliferating luminal edge of the neointima and the adventitial de-differentiated fibroblasts and may serve as a hallmark of this process. The possible functional consequences of this enzyme upregulation and its role in the response to vascular injury are suggested but remain to be determined.
Asunto(s)
Aciltransferasas/metabolismo , Angioplastia de Balón/efectos adversos , Traumatismos de las Arterias Carótidas/enzimología , Animales , Fibroblastos/enzimología , Inmunohistoquímica , Masculino , Modelos Animales , Músculo Liso Vascular/citología , Músculo Liso Vascular/enzimología , Ratas , Ratas Sprague-Dawley , Serina C-PalmitoiltransferasaRESUMEN
An RNA species has been identified whose nucleotide sequence is closely related to the mRNA encoding the murine interferon (IFN)-induced guanylate-binding protein-1 (mGBP1) and an mRNA encoding an isoprenylated protein that is constitutively expressed in various organs in the rat. Sequence analysis of the gene encoding this newly identified RNA reveals that in its 5'-region it is identical to a DNA fragment reported to represent the 5'-region of a gene termed mGBP2. In light of this homology, we term this newly identified gene product mGBP2. mGBP2 is inducible following IFN treatment in animals bearing Gbp1a alleles, in which mGBP1 is transcriptionally upregulated by IFN treatment, as well as in animals bearing Gbp1b alleles, in which mGBP1 is not induced in response to IFN treatment. The genomic organizations of the genes encoding mGBP1 and mGBP2 are similar, and the nucleotide sequences of their IFN-responsive-like elements and their relative locations are conserved. Gbp1 and Gbp2 map to a genetically indistinguishable site on the distal arm of chromosome 3.
Asunto(s)
Mapeo Cromosómico , Proteínas de Unión al ADN/genética , Proteínas de Unión al GTP , Genoma , Interferones/farmacología , Animales , Secuencia de Bases , Proteínas de Unión al ADN/biosíntesis , Datos de Secuencia Molecular , Prenilación de Proteína , Ratas , Homología de Secuencia de Ácido NucleicoRESUMEN
A cDNA encoding the human guanylate binding protein-1 (hGBP-1) was expressed in HeLa cells using a constitutive expression vector. Stably transfected clones expressing hGBP-1 exhibited resistance to the cytopathic effect mediated by both vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV) and produced less viral progeny than control cells following infection with these viruses. To study the role hGBP-1 plays in the IFN-mediated antiviral effect, cells were stably transfected with a construct expressing antisense RNA for hGBP-1. VSV infection of IFN-alpha-treated antisense RNA-expressing cells produced an amount of virus comparable to that produced in the parental cell line, while EMCV infection of the IFN-alpha-treated transfected cells and VSV and EMCV infection of the IFN-gamma-treated transfected cells produced far more virus than was produced in the parental cell line. These results demonstrate that GBP-1 mediates an antiviral effect against VSV and EMCV and plays a role in the IFN-mediated antiviral response against these viruses.