亚麻酸对赤潮异弯藻和米氏凯伦藻化感效应及细胞凋亡机制的研究

发布时间：2018-04-26 05:20

本文选题：亚麻酸 + 赤潮异弯藻　；参考：《曲阜师范大学》2015年硕士论文

【摘要】：近年来,有害赤潮在我国沿海和世界各地频繁发生,严重的影响了海洋生态系统、海洋水产系统稳定,甚至对海洋渔业和人类健康产生了巨大的威胁。有害赤潮已经成为海洋环境污染的问题之一,并引起了全球的关注。目前,对于有害赤潮的治理方法有很多种,但是,很多治理方法对于海洋系统具有一定的破坏性,所以,寻找一种高效、低影响的赤潮治理方法迫在眉睫。我们研究的重点是,依据前人研究的相生相克原理,利用大型海藻细胞内提取出的不饱和脂肪酸来抑制海洋赤潮微藻的生长。目前,有关化感物质的抑制机理研究主要有抗氧化系统、光合系统和胞外酶等。尤其是氧化协迫系统,因为氧化胁迫系统多于其他的影响因子关联在一起,但是化感物质是否直接作用于某种蛋白质或者说是否通过直接影响电子在叶绿体、线粒体中的传递而引起氧化胁迫的问题还有待于进一步的研究。另外,关于化感物质对于赤潮微藻的抑制是否是通过细胞凋亡的途径引起的研究鲜见报道。所以说,化感物质—不饱和脂肪酸对于赤潮微藻的氧化胁迫和细胞凋亡研究对于化感物质抑藻剂的开发和利用具有重要的意义。化感物质亚麻酸是在前期研究中从小珊瑚藻和鼠尾藻组织中分离鉴定出来的,具有很强的杀藻活性,本文进一步的探究了亚麻酸对赤潮异弯藻和米氏凯伦藻的抑制机理。通过研究亚麻酸对赤潮异弯藻和米氏凯伦藻氧化胁迫和细胞凋亡的研究来探讨亚麻酸对赤潮微藻的抑制机理。亚麻酸作为不饱和脂肪酸的一种,普遍存在在植物体中,是动植物生长发育必不可少的物质之一。我们通过研究亚麻酸对有害赤潮微藻氧化胁迫及细胞凋亡机制的影响来探究亚麻酸对于有害赤潮微藻的化感效应。赤潮异弯藻、米氏凯伦藻这两种藻是近年来有害赤潮形成的主要藻种,本文主要利用多种实验方法来探究不饱和脂肪酸—亚麻酸对赤潮异弯藻和米氏凯伦藻氧化胁迫和细胞凋亡机制的研究,从而取得了以下实验结果:1.亚麻酸对赤潮异弯藻和米氏凯伦藻生长量的影响亚麻酸对赤潮异弯藻和米氏凯伦藻生长量的影响呈现低浓度促进,高浓度抑制的趋势。2.亚麻酸对赤潮异弯藻和米氏凯伦藻抗氧化系统的影响亚麻酸作用赤潮异弯藻和米氏凯伦藻48h后,总的来说,随着亚麻酸浓度的增加,抗氧化酶(SOD、CAT、POD)的活性和抗氧化物质(抗坏血酸、GSH)都出现了先升高后下降的趋势。3.亚麻酸对赤潮异弯藻和米氏凯伦藻活性氧的影响亚麻酸作用赤潮异弯藻和米氏凯伦藻48h后,随着亚麻酸浓度的增加,羟基自由基、过氧化氢和ROS的含量逐渐的升高,超氧阴离子自由基含量先是升高,随后又出现了下降的趋势。4.亚麻酸对赤潮异弯藻和米氏凯伦藻氧化伤害的影响亚麻酸作用赤潮异弯藻和米氏凯伦藻48h后,随着亚麻酸浓度的增加,丙二醛的含量出现先升高后下降的趋势。通过丙二醛含量的变化来反应膜脂过氧化的程度。5.台盼蓝染色实验台盼蓝染色实验表明,亚麻酸作用赤潮异弯藻和米氏凯伦藻48h后,随着亚麻酸浓度的增加,藻细胞的成活率逐渐下降。6.亚麻酸对赤潮异弯藻和米氏凯伦藻细胞凋亡机制的影响(1)赤潮异弯藻和米氏凯伦藻的超微结构实验表明,亚麻酸处理赤潮异弯藻后,赤潮异弯藻的细胞出现细胞膜内陷,细胞器排列紊乱,部分质体界线不明显并开始溶解,液泡变大,有脂肪滴出现;亚麻酸处理米氏凯伦藻后,米氏凯伦藻细胞出现质壁分离,细胞内部细胞器排列紊乱,界线模糊,液泡增多,出现脂肪滴和黑色的蛋白颗粒。(2)线粒体膜电位实验表明,亚麻酸处理的赤潮异弯藻和米氏凯伦藻与未处理的对照组相比线粒体膜电位降低。(3)Caspase-3,9酶检测实验表明,亚麻酸处理的赤潮异弯藻和米氏凯伦藻与未处理的对照组相比,Caspase-3,9酶活性增高。(4)细胞色素c释放实验表明,亚麻酸处理的赤潮异弯藻和米氏凯伦藻与未处理的对照组相比,细胞色素c的含量增高。(5)Bcl-2实验表明,亚麻酸处理的赤潮异弯藻和米氏凯伦藻与未处理的对照组相比,Bcl-2的含量降低。
[Abstract]:In recent years, the harmful red tide has occurred frequently in China's coastal and all over the world, seriously affecting the marine ecosystem, the stability of the marine aquatic system, and even a great threat to the marine fisheries and human health. Harmful red tides have become one of the problems of marine environmental pollution and have aroused global concern. At present, the harmful red tide is concerned. There are many kinds of control methods, but many methods are destructive to the marine system. Therefore, it is imminent to find a high efficient and low impact method of red tide treatment. The growth of oceanic red tide microalgae. At present, the mechanisms of inhibition of allelochemicals are mainly antioxidant systems, photosynthetic systems and extracellular enzymes, especially oxidative stress systems, because the oxidative stress system is associated with other factors, but whether allelochemicals are directly connected to a certain protein or through straight. The problem of oxidative stress caused by the transmission of electrons in the chloroplasts and mitochondria remains to be further studied. In addition, there are few reports on whether the inhibition of allelochemicals to the red tide microalgae is caused by cell apoptosis. Therefore, the allelochemicals - unsaturated fatty acids are the threat to the oxidation of red tide microalgae The study of pressure and cell apoptosis is of great significance for the development and utilization of Allelochemical alga suppressor. Allelic linolenic acid is isolated and identified from the early studies of small coral algae and rat tail algae, and has strong algal activity. The mechanism. The inhibition mechanism of linolenic acid to red tide microalgae was investigated by studying the oxidative stress and apoptosis of linolenic acid to alga and Karen algae. Linolenic acid, one of the unsaturated fatty acids, is commonly found in plants and is one of the indispensable substances in the growth and development of animals and plants. The effects of acid on oxidative stress and cell apoptosis mechanism of harmful red tide microalgae are studied to explore the allelopathic effect of linolenic acid on harmful red tide microalgae. Two kinds of algae, Karen algae, are the main species of harmful red tides in recent years. This paper mainly uses a variety of experimental methods to explore the red tide of unsaturated fatty acid linolenic acid. Studies on the oxidative stress and cell apoptosis mechanism of alga and Karen algae, the following experimental results were obtained: 1. the effects of linolenic acid on the growth of alga and Karen alga in the red tide, the effect of linolenic acid on the growth of alga and Karen algae was low concentration, and the tendency of high concentration inhibition was.2. linolenic acid to red tide The effects of linolenic acid on the antioxidant system of alga and Karen Karen after the action of linolenic acid and Karen alga, in general, the activity of antioxidant enzymes (SOD, CAT, POD) and antioxidants (ascorbic acid, GSH) increased first and then decreased with the increase of linolenic acid concentration, and.3. linolenic acid was applied to the red tide ISO flexural algae and Michaelis. After the effect of linolenic acid on the effect of linolenic acid on 48h, the content of hydroxyl radicals, hydrogen peroxide and ROS increased gradually with the increase of linolenic acid concentration, the content of superoxide anion radicals increased first, and then the decreased potential of.4. linolenic acid was found in the oxygen of alga of red tide and Karen algae. The effect of linolenic acid on the effect of linolenic acid on the alga and Karen 48h, with the increase of the concentration of linolenic acid, the content of malondialdehyde first increased and then declined. The degree of malondialdehyde content changes to reflect the degree of membrane lipid peroxidation in.5. trypan blue staining experiment on the trypan blue dyeing experiment show that the flaxenic acid acts on the red tide After 48h, the survival rate of the algal cells decreased with the increase of linolenic acid concentration, and the effect of.6. linolenic acid on the apoptosis mechanism of the red tide isobent algae and Karen algae (1) the ultrastructural experiment of the red tide and Karen algae showed that the cells of the red tide ISO flexural algae were thin after the linolenic acid was at the red tide. In the cell membrane, the organelles were in disorder, the part of the plastid boundary was not obvious and began to dissolve, the vacuole became larger, and the fat droplets appeared. After linolenic acid treatment of Karen algae, the cells of Karen algae were separated, the internal organelles were arranged in disorder, the boundaries were blurred, the vacuoles increased, and the lipid droplets and black protein particles appeared. (2) mitochondrial membrane (2) membrane The potential test showed that the mitochondrial membrane potential decreased compared with the untreated control group. (3) the Caspase-3,9 enzyme test showed that the activity of Caspase-3,9 was higher than that of the untreated control group, compared with the untreated control group. (4) the experiment of cytochrome c release. Compared with the untreated control group, the content of cytochrome C increased in linolenic acid treated alga and Karen algae. (5) Bcl-2 experiment showed that the content of Bcl-2 was lower than that of untreated control group.

【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

【参考文献】