Evaluation of the Toxicity of Dichlorodiphenyl-trichloroethane (DDT) to the Expression Levels of Three Genes p53, rarα1, wnt in the Embryos and Liver of Fish Medaka Oryzias latipes

T. T. Huong, Nguyen Xuan Tòng
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Abstract

Three genes p53, rarα1, wnt and liver of Medaka fish tissue were selected to study the toxicity of DDT to fish embryo (8 days old) and adult Medaka fish (3 months old). TEM and real-time PCR methods were used to examine changes in structure of liver tissue and genetic expression. The TEM results recorded a clear difference between the control and the experimental sample. Adult Medaka fish exposed to 1µg/L DDT for 24 hours has a distorted liver cell nucleus with many hollow lipid particles, and the sinus-shaped meshes are clustered, interrupted; lysosome is not intact with many degenerative bubbles. The results of real-time PCR analysis showed that DDT affects the development, different and proliferation of cells in the embryonic stage and gene expression in adult stage. All three genes p53, rarα1 and wnt in fish embryos changed strongly and tended to be inhibited when exposuring with 1700µg/L DDT, the values ​​were recorded respectively 0.9; 4.9 and 5.4 times compared to the control sample (1 times). For adult fish, gene expression was lower than fish embryos with real-time PCR analysis values ​​for the three genes p53, rarα1 and wnt respectively 0.9 and 0.5; 0.36 and 0.09; 0.53 and 0.09 times after exposure to 1500 and 1700 µg/L DDT. These results demonstrated that gene expression is dependent on the developmental stage of the cell as well as on the dose of the chemical exposed. The development, cell differentiation/proliferation in the embryonic and adulthood stages affected differently to the cell gene expression.
二氯二苯三氯乙烷(DDT)对米鳉鱼胚胎和肝脏中p53、rarα1、wnt三个基因表达水平的毒性评价
本实验选取美达卡鱼组织的p53、rarα1、wnt和肝脏3个基因,研究DDT对8日龄美达卡鱼胚胎和3月龄美达卡鱼成鱼的毒性。采用透射电镜(TEM)和实时荧光定量PCR (real-time PCR)检测肝组织结构和基因表达的变化。透射电镜结果记录了对照和实验样品之间的明显差异。1µg/L滴滴涕作用24小时后,成鱼肝细胞核扭曲,有许多中空的脂质颗粒,窦状网簇状,中断;溶酶体不完整,有许多退行性气泡。实时荧光定量PCR分析结果显示,滴滴涕影响胚胎期细胞的发育、分化和增殖以及成虫期基因的表达。1700µg/L滴滴涕作用下,鱼胚中p53、rarα1和wnt三个基因均发生了强烈变化,并有被抑制的趋势,其值分别为0.9;与对照样品(1倍)相比,分别为4.9倍和5.4倍。在成鱼中,p53、rarα1和wnt三个基因的实时PCR分析值分别为0.9和0.5,表达量低于鱼胚;0.36和0.09;暴露于1500µg/L和1700µg/L DDT后分别为0.53和0.09倍。这些结果表明,基因表达依赖于细胞的发育阶段以及暴露的化学物质的剂量。胚胎期和成人期的发育、细胞分化/增殖对细胞基因表达的影响不同。
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