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dc.date.accessioned2013-03-12T08:52:59Z
dc.date.available2013-03-12T08:52:59Z
dc.date.issued2007en_US
dc.date.submitted2007-12-17en_US
dc.identifier.citationRuud, Øyvind. The effects of arterial shear on MAP kinase activation and expression of pro- and anti-inflammatory molecules in cultured porcine endothelial cells. Hovedoppgave, University of Oslo, 2007en_US
dc.identifier.urihttp://hdl.handle.net/10852/12196
dc.description.abstractVein grafting is complicated by high rates of stenosis due to the development of vascular inflammation, intimal hyperplasia and accelerated atherosclerosis. In contrast, arterial grafts are relatively resistant to these processes. Vascular inflammation is regulated by MAP kinases (including JNK and p38) and NF-ƒÛB signaling pathways which trigger endothelial expression of adhesion molecules (e.g. E-selectin, VCAM-1) and chemokines (e.g. IL-8, MCP-1). Blood flow over lumenal surfaces generates haemodynamic forces. Changes in flow have profound effects on the physiology of endothelial cells. This may influence disease processes that are associated with perturbations in flow, e.g. intimal thickening. Here we examined the effects of acute arterial shear stress (12 dynes/cm2) on pro-inflammatory activation of porcine aortic endothelial cells (PAEC) or porcine jugular vein endothelial cells (PJVEC). We also examined whether induction of anti-inflammatory transcripts accompanied the induction of pro-inflammatory mRNA. PAEC and PJVEC monolayers were plated onto gelatin-coated glass slides and cultured for 48h before experimentation. Cultures were then placed in a vacuum-held, parallel-plate flow chamber (Figure 2.2). Fluid was circulated through the chamber via a flow loop that held a constant flow rate, so that the shear stress imposed on the endothelial cells layer was 12 dynes/cm2 (equivalent to arterial flow) (Figure 2.1). Cells were cultured in static conditions or exposed to acute shear or TNF£\ (10ng/ml) for two and four hours or thirty and ninety minutes to look at gene expresssion and MAP kinase activation respectively. Transcript levels were quantified using reverse transcription, real-time quantitative PCR. The amount of each target gene was normalized by measuring cyclophilin mRNA levels. The effect of shear stress on pro-inflammatory MAP kinases and NF-£eB was assessed by western blotting for a phosphorylated form of JNK and p38, and nuclear translocation of p65. We observed that arterial shear stress induced high levels of IL-8, MCP-1 and E-selectin transcripts in PJVEC whereas PAEC were relatively resistant to the pro-inflammatory effects of shear stress (Figure 3.3 and Figure 3.4). To investigate the potential mechanism underlying the differential responses of PJVEC and PAEC we examined the effects of shear stress on the activation of NF-ƒÛB, JNK and p38. We observed that shear stress induced prolonged activation of JNK and p38 in PJVEC but only transient activation in PAEC. In contrast, the kinetics of NF-ƒÛB activation in response to shear were similar in both cell types (Figure 3.6 and Figure 3.7). Thus prolonged activity of MAP kinases may explain the hypersensitivity of PJVEC to shear stress mediated activation. Finally, we observed that anti-inflammatory transcripts (MKP-1 and XIAP) are induced by shear stress in PAEC but not in PJVEC (Figure 3.11), thus PAEC may resist pro-inflammatory activation by shear stress through inhibition of MAPK by molecules such as MKP-1 and XIAP. We suggest that the hypersensitivity of venous EC to the pro-inflammatory effects of shear stress may partly explain the susceptibility of vein grafts to inflammation and accelerated atherosclerosis.nor
dc.language.isoengen_US
dc.subjectendothelium inflammasjon atherosklerose intimial hyperplasi MAPK JNP p38 NFkBen_US
dc.titleThe effects of arterial shear on MAP kinase activation and expression of pro- and anti-inflammatory molecules in cultured porcine endothelial cellsen_US
dc.typeMaster thesisen_US
dc.date.updated2008-03-04en_US
dc.creator.authorRuud, Øyvinden_US
dc.subject.nsiVDP::568en_US
dc.identifier.bibliographiccitationinfo:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft.au=Ruud, Øyvind&rft.title=The effects of arterial shear on MAP kinase activation and expression of pro- and anti-inflammatory molecules in cultured porcine endothelial cells&rft.inst=University of Oslo&rft.date=2007&rft.degree=Hovedoppgaveen_US
dc.identifier.urnURN:NBN:no-18625en_US
dc.type.documentHovedoppgaveen_US
dc.identifier.duo68971en_US
dc.contributor.supervisorPaul Evans, Harald Thidemann Johansenen_US
dc.identifier.bibsys080352553en_US
dc.identifier.fulltextFulltext https://www.duo.uio.no/bitstream/handle/10852/12196/1/OeyvindxRuudxThesisx-xfinalx24.10.07.pdf


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