二十碳五烯酸对赤潮微藻氧化胁迫的影响及细胞凋亡机制的研究

发布时间：2018-01-30 13:40

  本文关键词： EPA 赤潮异弯藻 米氏凯伦藻 氧化胁迫 细胞凋亡机制　出处：《曲阜师范大学》2015年硕士论文　论文类型：学位论文

【摘要】：近年来,随着海洋工业经济活动的日益加剧,海洋环境污染成为当今全球关注的重要问题之一,而赤潮现象的频繁爆发,严重损害了海洋经济的可持续发展,给人类健康带来严重的危害。如何高效合理的治理赤潮问题成为科学家们关注的焦点。通过大型海藻分泌的化感物质处理赤潮问题是一种蓬勃发展的新型环境亲和型抑制方法,具有快速、高效、无二次污染等特点。从大型海藻中提取出来的不饱和脂肪酸来抑制赤潮微藻的生长是本文研究的重点。二十碳五烯酸(EPA)是不饱和脂肪酸的典型代表,普遍存在于水生动植物体内。本文通过观察EPA对赤潮微藻的生长以及氧化胁迫方面的影响,进一步对细胞的凋亡机制进行了探究,从新的视角阐明化感机制,为建立一套完善而实用的赤潮防治方法提供理论依据。赤潮异弯藻和米氏凯伦藻是海洋中比较常见的单细胞藻类,作为海洋中的初级生产者,是海洋食物链的关键所在。本文通过实验,分别从“个体水平—细胞水平—亚细胞水平—分子水平”上研究不饱和脂肪酸对赤潮异弯藻和米氏凯伦藻影响,取得了以下研究结果:1.EPA对赤潮异弯藻和米氏凯伦藻生长的影响EPA对赤潮异弯藻和米氏凯伦藻生长的影响均呈现出低浓度促进高浓度抑制的趋势。表现出明显的“低促高抑”现象。2.EPA对赤潮异弯藻和米氏凯伦藻氧化胁迫的影响(1)在EPA的作用下,赤潮异弯藻和米氏凯伦藻的SOD、CAT、POD、GSH、ASA均出现一定的应激活性,都表现出“低促高抑”的现象。(2)药物处理后,ROS水平、·OH、H2O2的水平也急剧上升,O2·-出现先上升后下降的趋势,这一现象可能与细胞内其它物质对O2·-的作用有关。(3)MDA含量先上升后下降,但下降趋势不是很显著,总体来说MDA的含量保持在一个较高的水平,其含量的变化体现了微藻膜系统受损伤的程度。(4)通过对照组可以看出,赤潮异弯藻和米氏凯伦藻本身的酶活性就不同。3.EPA对赤潮异弯藻和米氏凯伦藻细胞凋亡机制的影响(1)台盼蓝染色实验显示:EPA作用下,赤潮异弯藻和米氏凯伦藻的生长出现先上升后下降的趋势。(2)EPA作用下,赤潮异弯藻细胞出现严重变形,细胞膜破坏严重,外形改变,有凹陷,部分地方出现断裂,外膜出现溶解现象。而细胞内部的淀粉粒增多,液泡增多变大,质体出现空洞,基粒片层间隙变大,出现水肿现象,叶绿体外膜出现溶解现象。而米氏凯伦藻细胞出现明显的质壁分离现象,细胞壁逐渐溶解断裂,细胞外形发生严重改变,细胞内部叶绿体界限不清,出现溶解。(3)EPA处理赤潮藻后,赤潮异弯藻和米氏凯伦藻线粒体膜电位出现下降的现象。(4)EPA处理赤潮藻后,赤潮异弯藻和米氏凯伦藻细胞内的caspases-3,9含量均上升。(5)在药物作用下,赤潮异弯藻和米氏凯伦藻细胞内Cyt C从线粒体膜间歇释放出来。(6)在药物作用下,赤潮异弯藻和米氏凯伦藻细胞内Bcl-2家族蛋白受到抑制。
[Abstract]:In recent years, with the increasing economic activities of marine industry, marine environmental pollution has become one of the most important issues of global concern, and the frequent outbreak of red tide has seriously damaged the sustainable development of marine economy. How to manage red tide efficiently and reasonably has become the focus of scientists' attention. It is a booming new type of environment to deal with the problem of red tide by allelopathic substances secreted by macroalgae. Sum type inhibition method. Fast and efficient. No secondary pollution and other characteristics. The unsaturated fatty acids extracted from macroalgae to inhibit the growth of red tide microalgae is the focus of this study. 20 carbapentaenoic acid EPAs is a typical representative of unsaturated fatty acids. The effect of EPA on the growth of red tide microalgae and oxidative stress was observed to further explore the mechanism of cell apoptosis. The mechanism of allelopathy was elucidated from a new perspective to provide a theoretical basis for the establishment of a set of perfect and practical methods for the prevention and control of red tide. As the primary producer in the ocean, it is the key of the marine food chain. The effects of unsaturated fatty acids (unsaturated fatty acids) on the effects of unsaturated fatty acids (UFA) on Heterophyllum mirabilis and Kellenella mirabilis were studied from individual level, cell level, subcellular level and molecular level, respectively. The following results were obtained:. 1. The effects of EPA on the growth of Alternaria mirabilis and Kellenella mirabilis; 1. The effects of EPA on the growth of Alternaria mirabilis and Kellenella mirabilis showed a tendency of low concentration and high concentration inhibition. The effect of EPA on oxidative stress of Heterophyllum and Karen's algae in Red Tide. 1) under the action of EPA. Both the red tide and the SODCATCAT PODH ASA showed a certain stress activity, both of which showed the phenomenon of "low promotion and high inhibition" after drug treatment. The level of ROS, H _ 2O _ 2 and H _ 2O _ 2 also increased sharply, and O _ 2 路- showed a tendency of rising first and then decreasing. This phenomenon may be related to the effect of other substances in the cell on O2 路-. The content of MDA increased first and then decreased, but the decreasing trend was not very significant. In general, the content of MDA remained at a high level, and the change of the content reflected the degree of damage to the membrane system of microalgae. The effect of EPA on the apoptosis mechanism of Heteropodium mirabilis and Karen's alga. (1) trypan blue staining showed that under the action of: EPA. Under the action of EPA, the growth of the red tide and Karen's algae rose first and then decreased. Under the action of EPA, the cell membrane was seriously deformed, the cell membrane was destroyed seriously, the shape changed and there was a depression. In some places, rupture and dissolution appeared in the outer membrane. However, the number of starch granules in the cells increased, the vacuoles increased, the plastids became empty, the interlamellar gap of the granulosa became larger, and the phenomenon of edema appeared. The outer membrane of chloroplast dissolves, while the cytoplasm wall is separated obviously, the cell wall dissolves and breaks gradually, the appearance of the cell changes seriously, and the boundary of chloroplast inside the cell is not clear. After treated with EPA, the mitochondrial membrane potential of Alternaria mirabilis and Kellenella mirabilis decreased after the treatment of red tide alga with EPA. The contents of caspases-3t9 in the cells of both the red tide and Karen's alga were both increased. (5) under the action of the drug, the contents of caspases-3O9 in the cells were increased. Cyt C was released intermittently from mitochondria membrane. The Bcl-2 family proteins were inhibited by the drug treatment.
【学位授予单位】：曲阜师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X55;Q945.78

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