抗氧化酵素拮抗Thioacetamide與HCV core protein在斑馬魚早期胚胎發育中所誘導的氧化壓力

• Main Author: 王嘉蓉
• Other Authors: 耿全福
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/53564766883832633332

碩士 === 國立彰化師範大學 === 生物技術研究所 === 99 === Thioacetamide (TAA) is considered as one of the hepatotoxin. Hepatitis C virus (HCV) core protein (HCP) has been considered to be a significant risk factor in Hepatocellular carcinoma (HCC). But little is known about the toxic effect of TAA and HCP on animal embryos development, and whether HCP enhance the toxicity of TAA or not. First, we treated wild type zebrafish embryos with TAA. Our results showed that the survival rate were decreased to about 80% after treatment for 6 h. Does-dependent and time-course studies showed the LC50 was 24 mM for 84 h treatment. We also observed there are 45% of embryos were spinal column curving and 62% were edema in pericardial sac, and embryos with low melanin formation in skin and cannot hatch out from chorion after TAA treatment for 48 h. In addition, hydrogen peroxide was increased, nitric oxide was increased approximately 5-fold, SOD activity was decreased 24.5%, and oxidative damage marker include the levels of lipid peroxidation (LPO) were increased more than 2.6-fold after TAA-treated for 6 h when comparing with control. Utilizing the iNOS inhibitor-LCanavanine and increasing the level of antioxidant enzyme Prx-2 in embryos could effectively repressed oxidative stress which was induced by TAA, and then to reduced embryos death. Appearance of HCP transgenic fish embryos was nomal. But superoxide level was increased at 48 h , nitric oxide level was always more than wild type. Whereas LPO was obviously less than wild type. We then treated HCP transgenic zebrafish embryos with TAA. The survival rates were decreased to about 80% after treatment for 6 h. Does-dependent and time-course studies showed the LC50 was 24 mM for 60 h treatment. The teratogenic behavior were not only same with wild type embryostreated with TAA but also percentage was higher. Our datas showed that superoxide was increased at 6-24 h, SOD activity was highly inhibited 85.7% at 6 h, and LPO were increased 26.6% at 24 h after TAA treatment when comparing with HCP transgenic fish embryos. We demonstrated that HCP enhanced toxic of TAA to increase oxidative stress in early zebrafish embryonic development. Besides, we used microarray analysis to compare the global gene expression responses to TAA and HCP in zbrafish embryos. As result, hsp70l、hsp70、ZGC:174006 were up-regulated and nrp2b、epha4a、otx2、prdm1a、six3b、ZGC:136531與mef2cb were down-regulated after TAA treatment.