典型POPs对雄性翡翠贻贝毒理效应的蛋白组学和代谢组学研究

发布时间：2018-04-19 00:28

本文选题：翡翠贻贝 + 鳃　；参考：《海南大学》2015年硕士论文

【摘要】：滴滴涕(Dichlorodiphenyltrichloroethane, DDT)和苯并芘(Benzo(a)pyrene, BaP)均是典型持久性有机污染物(POPs),被认为是广泛存在的环境雌激素,由于其具有持久性、半挥发性或难挥发性、生物蓄积性、高毒性和远距离迁移性,而引起世界各国的广泛关注。本研究选择苯并芘和滴滴涕作为暴露污染物,以热带海洋重要模式生物翡翠贻贝(Perna viridis)为研究对象,采用代谢组学和蛋白质组学技术,探究两种典型POPs单独及联合作用对雄性翡翠贻贝鳃及雄性生殖腺的毒理效应,揭示其毒理机制,为早期预警监测两种污染物和风险评价提供科学依据。 1.BaP和DDT及其联合作用对雄性翡翠贻贝鳃组织的代谢组学和蛋白质组学研究通过对雄性翡翠贻贝进行单独暴毒BaP10μg/L、 DDT10μg/L及联合暴毒BaP10μg/L+DDT10μg/L,各组均处理7天后,基于核磁共振技术的代谢组学分析结果表明：BaP单独暴露主要影响雄翡翠贻贝鳃组织的渗透压调节,而DDT单独暴露主要影响雄翡翠贻贝鳃组织的渗透压调节和能量代谢过程。BaP和DDT联合暴毒后雄性翡翠贻贝鳃组织未检测到明显的代谢物变化。利用基于双向电泳技术和质谱检测技术相结合的方法来研究雄性翡翠贻贝鳃组织的蛋白质表达谱的变化,结果显示：BaP暴露使雄翡翠贻贝鳃组织发生变化的蛋白质涉及细胞凋亡,蛋白质消化和能量代谢等生物过程；DDT暴露使雄翡翠贻贝鳃组织变化的蛋白质涉及氧化应激,蛋白质消化,能量代谢,细胞骨架和结构等生物过程；BaP和DDT联合暴露后,雄翡翠贻贝鳃组织响应的蛋白质涉及氧化应激,蛋白质消化,能量代谢,细胞生长和细胞凋亡,细胞骨架和结构等生物过程。 2.BaP和DDT及其联合作用对雄性翡翠贻贝生殖腺的代谢组学和蛋白质组学研究对翡翠贻贝进行BaP10μg/L, DDT10μg/L及BaP10μg/L+DDT10μg/L暴毒7天后,基于核磁共振技术的代谢组学分析结果表明：BaP单独暴露组雄翡翠贻贝的生殖腺没有明显的代谢物变化。而DDT单独暴露和BaP与DDT联合暴露均主要影响雄翡翠贻贝生殖腺组织的渗透压调节和能量代谢过程。与DDT单独暴露组相比,联合暴露组支链氨基酸、丙氨酸,谷氨酸,甘氨酸、乙酰乙酸、琥珀酸、龙虾肌碱、精氨酸、未知代谢物(3.53ppm)和ATP均发生变化,且变化趋势相同。表明BaP和DDT联合暴毒组的代谢物变化主要是由DDT引起的,DDT对雄翡翠贻贝生殖腺代谢物的影响较大。基于双向电泳结合质谱检测技术的蛋白质组学分析结果表明：BaP暴露后雄翡翠贻贝生殖腺响应的蛋白质的功能涉及信号传导、蛋白质分解代谢、物质和能量代谢、氧化应激、精子的发生和细胞凋亡等生物学过程；DDT暴露后雄翡翠贻贝生殖腺响应的蛋白质功能涉及细胞形成、蛋白质转运、能量代谢、转录因子、细胞骨架、氧化应激、信号传导、和细胞凋亡等生物学过程：BaP和DDT联合暴露后,雄翡翠贻贝生殖腺变化的蛋白质涉及细胞骨架、细胞形成、氧化应激、核苷酸代谢、转录因子、精子的发生、蛋白质转运、物质和能量代谢、信号传导、和细胞凋亡等生物学过程。该研究表明,联合蛋白质组学和代谢组学的方法可以更好地解释环境污染物对生物体的毒理机制。
[Abstract]:DDT (Dichlorodiphenyltrichloroethane, DDT) and benzo pyrene (Benzo (a) pyrene, BaP) are typical persistent organic pollutants (POPs), is considered to be the environmental estrogens widely exist, because of its persistence, semi volatile or non-volatile, bioaccumulation and high toxicity and long range transport, and caused widespread concern all over the world. This research chooses benzopyrene and DDT as exposure to tropical marine pollutants, an important model organism in mussel (Perna viridis) as the research object, using metabonomics and proteomics research, two typical POPs single and combined effects of toxicological effects on male mussel gills and male gonads, revealing its toxicological mechanism, provide scientific basis for early warning and risk assessment of two kinds of pollutants.
Metabolomics and proteomics of 1.BaP and DDT and their combined effects on the gill tissue of male jadeite mussel
By BaP10 g/L alone exposure toxicity on male mussel, DDT10 g/L and BaP10 g/L+DDT10 combined with drug exposure groups were g/L, 7 days after treatment, metabolic NMR analysis results show that: Based on the penetration of BaP exposure mainly affects male mussel gill tissue osmotic pressure regulation, while DDT alone exposure the main effects of male mussel gill tissue pressure regulation and energy metabolism of.BaP and DDT combined with drug exposure after male mussel gill tissue was not detected metabolites change significantly. The change of protein expression profile method, two-dimensional electrophoresis and mass spectrometry detection technology based on the combination of male mussel gill tissue showed that: BaP exposure to male mussel gill tissue changes of proteins involved in apoptosis, protein digestion and energy metabolism and other biological processes; DDT exposure to male mussel gill Histological changes of proteins involved in oxidative stress, protein metabolism, energy metabolism, cytoskeleton structure and biological processes; BaP and DDT after exposure, the male mussel gill tissue response proteins involved in oxidative stress, protein metabolism, energy metabolism, cell growth and apoptosis, cytoskeleton structure and biological processes.
Metabolic and proteomic studies of 2.BaP and DDT and their combined effects on the reproductive glands of the male jadeite Mytilus edulis
BaP10 g/L of DDT10 g/L and mussel, BaP10 mu g/L+DDT10 mu g/L 7 days after drug exposure, metabolic NMR analysis results show that: BaP based on single exposure group of male genital glands of Perna viridis NO metabolites change obviously. While DDT alone exposure and BaP and DDT joint exposure were main influence the male gonad tissue penetration of mussel pressure regulation and energy metabolism. Compared with DDT alone exposure group, combined exposure group of branched chain amino acids, alanine, glutamic acid, glycine, acetoacetate, succinate, homarine, arginine, unknown metabolites (3.53ppm) and ATP were changed, and have the same trend. That metabolite the changes of BaP and DDT combined with drug exposure group is mainly caused by DDT, the effect of DDT on male reproductive gland mussel metabolites greatly. Two dimensional electrophoresis combined with mass spectrometry detection study based on the results of the analysis BaP showed that after exposure to the male gonad response of mussel protein function involving signal transduction, protein metabolism, energy metabolism, oxidative stress, and cell apoptosis after exposure to DDT sperm; male genital gland response of mussel protein functions involved in cell formation, protein transport, energy metabolism, transcription factor, cytoskeleton, oxidative stress, signal transduction, and cell apoptosis in BaP and DDT after exposure, the protein changes of the gonads of the male mussels involved in cytoskeleton, cell formation, oxidative stress, nucleotide metabolism, transcription factor, spermatogenesis, protein transport, material and energy metabolism. Signal transduction, and cell apoptosis. The study shows that the methodology and metabonomics combined with proteomics can better explain the environmental pollutants on organisms The mechanism of toxicology.

【学位授予单位】：海南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X171.5

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