碩士 / 輔仁大學 / 生命科學系碩士班 / 103 / Abstract
Herpes simplex virus type 1 (HSV-1; Herpesviridae) is an enveloped DNA virus and a risk factor for Alzheimer’s disease (AD). Polygonum multiflorum is applied for anti-aging in Chinese medicine. In the present study, anti-HSV-1 activity of P. multiflorum extracts (PM) and their effects on AD markers including amyloid precursor protein (APP) cleavage and Tau protein hyperphosphorylation in SH-SY5Y cells induced by HSV-1 infection, were evaluated. The results indicated that (1) PM blocked HSV-1 replication in Vero E6 cells in a dose-dependent manner with IC50 108 ±13.8 μg/ml. The inhibitory effect of PM was not related to direct cytotoxicity; (2) In the time of addition experiment, PM added at 0 to 8 hr postinfection (p.i.) reduced HSV-1 replication in Vero E6 cells; (3) To elucidate whether PM affected HSV-1 adsorption, the cells were pretreated with PM for 1 hr at 4 °C then infected with HSV-1, the virus titers were determined by plaque forming assay. The data showed that PM had 20% inhibitory activity on virus replication; (4) To determine whether PM interfered HSV-1 structure, the viral particles were treated with PM for 15 min to 60 min at 37 °C and virus titers were analyzed plaque forming assay. The results indicated that HSV-1 replication was blocked after treated with PM for 15 min to 1 hr; (5) The data from Western blotting demonstrated that HSV-1 late proteins such as glycoprotein B (gB), gC, and gG and immediated-early proteins including infected cell protein 0 (ICP0) and ICP4 protein expressions in Vero E6 cells were significantly attenuated by PM; (6) PM also inhibited HSV-1 replication in neuron cells SH-SY5Y; (7) Furthermore, to analyze whether HSV-1 infection affected APP cleavage and Tau protein hyperphosphorylation in neuron cells, SH-SY5Y cells were infected with HSV-1 and APP cleavage and Tau protein hyperphosphorylation were determined by Western blotting. The results indicated that F35 fragments cleavaged from APP (APP-F35) and hyperphosporylation of Tau protein could be induced by HSV-1 at 12 to 24 hr p.i.; (8) Effects of PM on AD markers expression in HSV-1-infected SH-SY5Y cells were analyzed with Western blotting. The results showed that PM decreased APP-F35 productions and levels of Tau protein hyperphosphorylation in the cells induced by HSV-1. Based on this study, I concluded that PM blocked HSV-1 replication in Vero E6 cells and SH-SY5Y cells. The inhibitory action mechanisms were related to blocking of HSV-1 adsorption, interruption of virus structure, and inhibition of virus immediated-early and late proteins expression. PM also decreased AD markers production in neuron cells induced by HSV-1. These data could be as evidences for development new antiviral drugs from PM and PM applied in AD prevention and treatment.
| Identifer | oai:union.ndltd.org:TW/103FJU00105015 |
| Date | January 2014 |
| Creators | Huang Shin Hui, 黃詩惠 |
| Contributors | Kuo,Yuh-Chi, Tsai, Wei-Jern, 郭育綺, 蔡維人 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 89 |
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