酞酸酯类对果蝇的毒效应

【作者】 厉曙光；

【导师】 金泰廙；

【作者基本信息】 复旦大学 ， 卫生毒理学， 2006， 博士

【摘要】 目的：研究DEHP和DBP—DOP等酞酸酯类化合物对果蝇寿命和体内抗氧化酶系统的影响，观察DEHP、DBP、DOP对果蝇生殖系统损害的组织病理学结果和遗传毒性，以及用基因芯片筛选DEHP诱导果蝇脑组织差异表达基因等，探讨其对黑腹果蝇的多种毒效应。方法：进行果蝇生存试验研究DEHP、DBP、DOP对果蝇寿命的影响；通过对果蝇经口喂饲DEHP后体内总SOD、CuZn-SOD活力及MDA含量的测定，研究DEHP对果蝇体内脂质过氧化反应的作用；对黑腹果蝇进行亚急性毒性试验，研究DEHP对果蝇生殖系统的损伤；利用高通量的基因芯片技术获得DEHP诱导果蝇脑组织的基因差异表达谱，从基因转录水平初步探讨DEHP缩短果蝇寿命的毒性作用机制；采用伴性隐性致死试验(SLRL)研究了DBP、DOP对果蝇生殖细胞的遗传毒性。结果：①DEHP和DBP—DOP可使果蝇半数死亡时间提前，200mg／L高剂量组雌雄果蝇分别较对照组提前12d及10d(P＜0.01或P＜0.05)，呈时间-依赖关系，并明显缩短果蝇的平均寿命和平均最高寿命(P＜0.01)，表明DEHP和DBP—DOP可使果蝇寿命缩短。②生化酶学检测发现DEHP染毒组果蝇体内总SOD和CuZn-SOD活力较对照组低(P＜0.01或P＜0.05)，并可使果蝇体内MDA含量增高(P＜0.01或P＜0.05)，提示DEHP可抑制果蝇体内SOD活力，促进脂质过氧化反应。③DEHP和DBP可对雄性黑腹果蝇生殖系统造成可逆性损伤。④DEHP诱导果蝇脑组织差异表达的基因片段共计35个：上调的基因有20个，包括CYP6a2，CYP6a8，CYP6w1，CYP4p1，Ugt36Bb，GST；下调的基因有15个，包括Mst57Da，Mst57Db，Acp95EF，Os-E，Os-C，Pbprp3，Pbprp1，a10等。⑤伴性隐性致死试验的结果表明；DBP-DOP对果蝇生殖细胞具有遗传毒性；果蝇的致突变率和不育率升高。结论：DEHP、DBP、DOP等对黑腹果蝇有一定的毒性作用，对人类可能产生潜在危害。更多还原

【Abstract】 [Objective] To study the effect of phthalates including DEHP and DBP-DOP on the lifespan, anti-oxydative system, reproductive system, genotoxicity, and neuotoxicity of Drosophila melanogaste. [Methods] ① Effect of DEHP and DBP-DOP on the lifespan of drosophila. was studied by the longevity test ②Determine the activity of total-SOD and CuZn-SOD and the concentration of MDA of the drosophila fed by DEHP orally, thus to study its impact on the lipid peroxidation in vivo.. ③Use subacute toxicity test to study the damage of DEHP on the reproductive system of drosophila. ④To determine the effect of DEHP on differential gene expression of the brain tissue by the use of RT-PCR （gene chips） thus to study its toxicity on drosophila. ⑤The genotoxicity of DBP-DOP on the germ cells of drosophila. was studied by The sex-linked recessive lethal （SLRL） test The result of sex-linked recessive lethal （SLRL） test shows genotoxicity to reproductive cells in drosophila induced with DBP-DOP. [Results] ①200mg/L dosage of DEHP and DBP-DOP shortened respectively the half lethal time (LT₅₀) of drosophila by 12d with the female and 10d with the male all compared with control（p<0. 01 or 0. 05）; a time-dependence relationship was shown. Also, mean and max of the lifespan shortened （p<0. 01） . So DEHP and DBP-DOP shortened respectively the lifespan of drosophila. ②Activity of total-SOD and CuZn-SOD of drosophila fed by DEHP was weaker than control （p<0.01 or 0.05） , however, concentration of MDA increased （p<0. 01 or 0.05） , thus to imply that DEHP inhibited the activity of SOD, promoted reaction of lipid peroxidation in vivo. ③DEHP and DBP caused respectively reversible damage to the reproductive system in drosophila. （4）35 fragments were induced by DEHP in the study of differential gene expression of brain tissue. 20 of them were upregulated,including CYP6a2, CYP6a8, CYP6wl, CYP4p1, Ugt36Bb, GST, Carboxylesterase, Glycine N-methyltransferase, Oxidoreductase, Est10, Jheh1, etc. with 8 new findings. 15 fragments were downregulated, including Mst57Da, Mst57Db, Acp95EF, Os-E, Os-C, Pbprp3, Pbprp1, a10, etc. with 7 new findings. Those genes involved in the functions of signal conduct, cell metabolism, growth, tissue differentiation and transcription factors with some for further comprehension.⑤ The result of sex-linked recessive lethal （SLRL） test shows genotoxicity to reproductive cells in drosophila induced with DBP-DOP and promote both rates of mutagen and sterility respectively. [Conclusion] DEHP, DBP and DOP have toxic effect on Drosophila melanogaster indicating potential hazards on human-being.更多还原