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標題: 豬瘟病毒感染內皮細胞對一氧化氮的影響
The Effect of Classical Swine Fever Virus (CSFV) on Nitric Oxide (NO) in Porcine Aortic Endothelial Cells (PAECS)
作者: 王之仰
關鍵字: 基礎研究;Classic swine fever virus(CSFV);畜牧獸醫類;豬瘟病毒;一氧化氮;一氧化氮合成?;nitric oxide;nitric oxide synthase

Classic swine fever (CSF) is a contagious disease of domestic pigs with high severity and mortality. It still causes a significant loss to pig industry in Taiwan even an extensive vaccination plan is implemented. Classic swine fever virus (CSFV) is a member of Pestivirus genus of Flaviridae. In acute and subacute cases, severe haemorrhagic diathesis and petechiation in multiple tissues and organs are derived from exaggerated inflammatory and immune responses and increased procoagulant activities. Vasculitis triggered by damages of endothelial cells when CSFV replicates inside constitutes the pathogenic basis of CSF. Nitric oxide (NO) identified as a vessel relaxing factor, a neurotransmitter, and a host defense mediator is generated from both a constitutive NO synthase (cNOS) and an inducible NO synthase (iNOS). Studies have indicated NO plays a role in both protective and cytotoxic responses in viral infections. NO produced by infected cells inhibited the replication of vaccinia virus and retroviruses. The highoutput NO from the up-regulated NO synthases followed cells infected with rabies virus and reovirus caused severe inflammations leading to tissue damages. A comprehensive study using culture cells for two years is proposed herein. Porcine aortic endothelial cells (PAEC) are infected with CSFV at different dosage and time dependent manners. The expression profiles and spatial localizations of cNOS and iNOS are checked by high resolution confocal laser scanning microscope. The expression levels and degradation rates of cNOS and iNOS are determined by semi-quantitative RT-PCR and Western blotting. The amount of nitric oxide and promoter activities of cNOS and iNOS are measured. Furthermore, the phosphorylation status of cNOS is examined. Selective inhibitors are used to determine the roles of PI 3-K/Akt pathway, Src protein, and intracellular calcium ions in eNOS phosphorylation. The expression of iNOS through MAPK pathway is analyzed. With past experiences in studies, this plan is surely to be conducted properly. This proposal will improve our understanding about molecular pathogenesis of CSFV and potentially contributes to develop a novel strategy for future prevention and therapy.
其他識別: NSC97-2313-B005-047-MY2
Appears in Collections:獸醫學系所

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