虎杖對EB病毒溶裂循環的抑制作用

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Title:	虎杖對EB病毒溶裂循環的抑制作用 Inhibition of the Epstein-Barr virus lytic cycle by Polygonum cuspidatum
Authors:	黃靖雯 Ching-Wen Huang
Contributors:	林翠品葉東柏嘉南藥理科技大學：生物科技研究所
Keywords:	白黎蘆醇 EB病毒大黃素虎杖抗病毒 EBV resveratrol emodin Polygonum cuspidatum antiviral activity
Date:	2008
Issue Date:	2009-03-11 11:42:09 (UTC+8)
Abstract:	虎杖是常使用之藥草，具有降低膽固醇、抗病毒、抗菌、抗血管新生的活性。本研究的目的是探討虎杖根之乙醇萃取物及其主要成分，大黃素與白黎蘆醇抑制EB病毒溶裂循環的效應。本研究以trypan blue及 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide方法分析細胞存活率；以西方墨點法、免疫螢光法及流式細胞儀分析EB病毒溶裂蛋白質－Rta、Zta及EA-D的表現；以Real-time PCR方法分析病毒顆粒的釋放；以螢光素酶分析病毒溶裂循環基因的轉錄。結果顯示虎杖根之乙醇萃取物及大黃素對於P3HR1細胞的CC50分別為為67.47 g/ml及11.46 M。虎杖根之乙醇萃取物、大黃素與白黎蘆醇分別在 25 g/ml、9.37 M及55 時具有抑制EB病毒溶裂蛋白質Rta、Zta及EA-D的表現。也分別降低66.70%、26.68%及54.21%病毒顆粒量。虎杖根之乙醇萃取物在濃度12.5 g/ml時對病毒溶裂循環早期基因BRLF1及BZLF1轉錄抑制能力分別為35.56%及45.36%；大黃素在濃度9.37 M時的抑制能力分別為3.21%及、31.17%；而白黎蘆醇在濃度6.88 M時降低BRLF1及BZLF1轉錄能力分別為88.11%及92.04%。綜合本研究發現虎杖根之乙醇萃取物及其主要成分大黃素與白黎蘆醇具有抑制EB病毒早期基因的轉錄，並且影響EB病毒溶裂蛋白質的表現，進而抑制病毒顆粒的產生。 Polygonum cuspidatum is a medicinal plant, with the cholesterol-lowering, antiviral, antibacterial and antiangiogenesis activities. This study is to investigate the inhibitory effects of the ethanolic extract of Polygonum cuspidatum root (PcE) and its main compnents, emodin and resvertrol, on Epstein-Barr virus lytic cycle. Trypan blue exclusion test and MTT [3-(4,5- dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] method are used to analyze the cell viability. Immunoblot, indirect immunofluorescence and flow cytometry analyze were used to investigate the expression of Epstein-Barr virus (EBV) lytic proteins, Rta, Zta, and EA-D; Real-time quantitative PCR was used to measure the EBV virion production; Tansient transfection analysis was followed to assess the transcription of lytic genes, including BRLF1 and BZLF1. Results showed that the PcE and emodin could influence the viability of P3HR1 cell, CC50 were 67.47 g/ml and 11.46 M, respectively. PcE, emodin and resvertrol at concentration of 25 g/ml, 9.37 M and 55 M, respectively, could inhibit the expression of EBV lytic proteins, including Rta, Zta, and EA-D; and reduce the virion production for 66.70%, 26.68%, and 54.21%, respectively. Furthermore, PcE, emodin and resvertrol inhibited the transcription of BRLF1 and BZLF1 by 35.56% and 45.36% at 12.5 g/ml; by 3.21% and 31.17% at 9.37 M; by 88.11% and 92.04% at 6.88 M, respectively. This study found that PcE, emodin and resvertrol could inhibit the transcription of lytic genes of EB virus, in turn, affected its lytic proteins expression. This inhibitory ability might impedes the production of mature viral particle.
Relation:	校內一年後公開，校外永不公開
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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