本文關鍵詞： EPA 赤潮異彎藻 米氏凱倫藻 氧化脅迫 細胞凋亡機制　出處：《曲阜師范大學》2015年碩士論文　論文類型：學位論文

【摘要】：近年來,隨著海洋工業(yè)經(jīng)濟活動的日益加劇,海洋環(huán)境污染成為當今全球關注的重要問題之一,而赤潮現(xiàn)象的頻繁爆發(fā),嚴重損害了海洋經(jīng)濟的可持續(xù)發(fā)展,給人類健康帶來嚴重的危害。如何高效合理的治理赤潮問題成為科學家們關注的焦點。通過大型海藻分泌的化感物質(zhì)處理赤潮問題是一種蓬勃發(fā)展的新型環(huán)境親和型抑制方法,具有快速、高效、無二次污染等特點。從大型海藻中提取出來的不飽和脂肪酸來抑制赤潮微藻的生長是本文研究的重點。二十碳五烯酸(EPA)是不飽和脂肪酸的典型代表,普遍存在于水生動植物體內(nèi)。本文通過觀察EPA對赤潮微藻的生長以及氧化脅迫方面的影響,進一步對細胞的凋亡機制進行了探究,從新的視角闡明化感機制,為建立一套完善而實用的赤潮防治方法提供理論依據(jù)。赤潮異彎藻和米氏凱倫藻是海洋中比較常見的單細胞藻類,作為海洋中的初級生產(chǎn)者,是海洋食物鏈的關鍵所在。本文通過實驗,分別從“個體水平—細胞水平—亞細胞水平—分子水平”上研究不飽和脂肪酸對赤潮異彎藻和米氏凱倫藻影響,取得了以下研究結果:1.EPA對赤潮異彎藻和米氏凱倫藻生長的影響EPA對赤潮異彎藻和米氏凱倫藻生長的影響均呈現(xiàn)出低濃度促進高濃度抑制的趨勢。表現(xiàn)出明顯的“低促高抑”現(xiàn)象。2.EPA對赤潮異彎藻和米氏凱倫藻氧化脅迫的影響(1)在EPA的作用下,赤潮異彎藻和米氏凱倫藻的SOD、CAT、POD、GSH、ASA均出現(xiàn)一定的應激活性,都表現(xiàn)出“低促高抑”的現(xiàn)象。(2)藥物處理后,ROS水平、·OH、H2O2的水平也急劇上升,O2·-出現(xiàn)先上升后下降的趨勢,這一現(xiàn)象可能與細胞內(nèi)其它物質(zhì)對O2·-的作用有關。(3)MDA含量先上升后下降,但下降趨勢不是很顯著,總體來說MDA的含量保持在一個較高的水平,其含量的變化體現(xiàn)了微藻膜系統(tǒng)受損傷的程度。(4)通過對照組可以看出,赤潮異彎藻和米氏凱倫藻本身的酶活性就不同。3.EPA對赤潮異彎藻和米氏凱倫藻細胞凋亡機制的影響(1)臺盼藍染色實驗顯示:EPA作用下,赤潮異彎藻和米氏凱倫藻的生長出現(xiàn)先上升后下降的趨勢。(2)EPA作用下,赤潮異彎藻細胞出現(xiàn)嚴重變形,細胞膜破壞嚴重,外形改變,有凹陷,部分地方出現(xiàn)斷裂,外膜出現(xiàn)溶解現(xiàn)象。而細胞內(nèi)部的淀粉粒增多,液泡增多變大,質(zhì)體出現(xiàn)空洞,基粒片層間隙變大,出現(xiàn)水腫現(xiàn)象,葉綠體外膜出現(xiàn)溶解現(xiàn)象。而米氏凱倫藻細胞出現(xiàn)明顯的質(zhì)壁分離現(xiàn)象,細胞壁逐漸溶解斷裂,細胞外形發(fā)生嚴重改變,細胞內(nèi)部葉綠體界限不清,出現(xiàn)溶解。(3)EPA處理赤潮藻后,赤潮異彎藻和米氏凱倫藻線粒體膜電位出現(xiàn)下降的現(xiàn)象。(4)EPA處理赤潮藻后,赤潮異彎藻和米氏凱倫藻細胞內(nèi)的caspases-3,9含量均上升。(5)在藥物作用下,赤潮異彎藻和米氏凱倫藻細胞內(nèi)Cyt C從線粒體膜間歇釋放出來。(6)在藥物作用下,赤潮異彎藻和米氏凱倫藻細胞內(nèi)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.
【學位授予單位】：曲阜師范大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：X55;Q945.78

【參考文獻】

中國期刊全文數(shù)據(jù)庫 前9條

1 王曉雯;;赤潮的預報及治理[J];環(huán)境科學與管理;2012年S1期

2 張冬鵬,武寶王干;幾種赤潮藻對溫度、氮、磷的響應及藻間相互作用的研究[J];暨南大學學報(自然科學與醫(yī)學版);2000年05期

3 李春強;劉志昕;黎娟華;廖文彬;孫海彥;趙平娟;于曉玲;彭明;;紅樹植物化感作用對中肋骨條藻生長的影響[J];熱帶作物學報;2009年06期

4 胡廷尖;王雨辰;陳豐剛;嚴力蛟;練青平;劉士力;;鳳眼蓮對銅綠微囊藻的化感抑制作用研究[J];水生態(tài)學雜志;2010年06期

5 黃長江;赤潮研究及其展望[J];生命科學;1999年S1期

6 李靜巖;Caspase-3與腫瘤關系的研究[J];醫(yī)學綜述;2005年05期

7 肖艷;呂回;;高氧對新生大鼠肺細胞色素C和caspase-3表達及細胞凋亡的影響[J];醫(yī)學綜述;2011年13期

8 張庭廷;吳安平;何梅;陳傳平;聶劉旺;;酚酸類物質(zhì)對水華藻類的化感作用及其機理[J];中國環(huán)境科學;2007年04期

9 李晶,閻秀峰,祖元剛;低溫脅迫下紅松幼苗活性氧的產(chǎn)生及保護酶的變化[J];植物學報;2000年02期

中國碩士學位論文全文數(shù)據(jù)庫 前1條

1 張智慧;Livin在非小細胞肺癌中的表達及與Caspase-3和PTEN的關系[D];鄭州大學;2006年

，

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