【摘要】：隨著經(jīng)濟的迅速發(fā)展,人類的醫(yī)療衛(wèi)生條件不斷改善,多達3000種藥物被用于醫(yī)療,并在環(huán)境中積累。由于污水處理廠的設備并不能將這些有機藥物完全去除,因此這些有機藥物被排放到水環(huán)境中,不斷滲透到地表水、地下水和沉積物中,使其在環(huán)境中的分布具有全球普遍性。其中,雙氯芬酸是一種最常用的、療效最好的非甾類鎮(zhèn)痛消炎藥(NSAIDs),被廣泛用于臨床治療中。由于雙氯芬酸藥品的療效好、用量大,被無節(jié)制地排放到環(huán)境中,并危及到了生態(tài)系統(tǒng)的穩(wěn)定性�，F(xiàn)在我們已知,雙氯芬酸能嚴重影響植物的生長發(fā)育,但是其抑制植物生長的作用機制至今尚不清楚。為此,本研究利用煙草BY-2細胞為材料,探討雙氯芬酸對煙草BY-2細胞的呼吸代謝及其線粒體功能的影響,利用Oxytherm氧電極(Hansatech,英國)測定不同濃度的雙氯芬酸鈉即時處理的煙草BY-2細胞的總呼吸,測定不同處理時間的煙草BY-2細胞的總呼吸、細胞色素途徑(COX)和交替氧化酶(AOX)途徑呼吸以及線粒體的狀態(tài)呼吸,測定不同呼吸途徑抑制劑處理的細胞呼吸,并利用稱量細胞干重的方法更直觀的驗證雙氯芬酸鈉對煙草BY-2細胞生長的抑制。進而,闡明雙氯芬酸抑制煙草BY-2細胞生長的作用機理,為闡明雙氯芬酸抑制植物生長的機理提供理論依據(jù)。本研究結果表明,0.2 mmol·L-1雙氯芬酸處理24h后,就顯著抑制了煙草BY-2細胞的生長,引起細胞內(nèi)活性氧(ROS)的爆發(fā)和積累,并導致煙草BY-2細胞死亡。并且,研究發(fā)現(xiàn)一定濃度的雙氯芬酸對煙草BY-2細胞的呼吸有即時的抑制作用,且隨著雙氯芬酸的濃度增加,對細胞呼吸作用的抑制程度也增加,當雙氯芬酸的濃度達到0.2mmol·L-1時,它對煙草BY-2細胞呼吸作用的抑制程度達到最大。此外,在雙氯芬酸對煙草BY-2細胞離體線粒體的直接作用的研究中,我們發(fā)現(xiàn)雙氯芬酸通過抑制細胞線粒體呼吸電子傳遞鏈上復合體II、III和IV的活性,導致線粒體中細胞色素氧化酶(COX)和交替氧化酶(AOX)參與的兩條呼吸電子傳遞途徑均受到不同程度的抑制作用,進而反饋抑制了細胞的糖酵解(EMP)、三羧酸循環(huán)(TCA)和戊糖磷酸途徑(PPP)三個碳代謝途徑。并且,雙氯芬酸對線粒體I、III、IV三個狀態(tài)的呼吸均有抑制作用,導致呼吸控制率(RCR)降低。而且,雙氯芬酸也能導致線粒體發(fā)生腫脹,并能降低線粒體膜磷脂的聚集程度,破壞了線粒體的完整性,導致跨膜質(zhì)子梯度形成受阻,進而造成ATP合成受抑。因此,我們認為煙草BY-2細胞中經(jīng)過線粒體呼吸電子傳遞鏈上復合體III和IV的電子傳遞被抑制是雙氯芬酸抑制細胞呼吸的一個主要原因。本研究證明了雙氯芬酸對線粒體呼吸鏈上復合體II,III和IV活性的抑制是它抑制細胞呼吸的主要原因,這就導致了線粒體代謝發(fā)生紊亂。線粒體能量代謝和物質(zhì)代謝的紊亂進一步造成細胞內(nèi)活性氧(ROS)爆發(fā)和積累,這些都是雙氯芬酸抑制煙草BY-2細胞生長、導致細胞死亡的重要原因。
[Abstract]:With the rapid development of the economy, human health conditions have been improving, as many as 3,000 drugs have been used for medical treatment and are accumulated in the environment. Since the equipment of the sewage treatment plant cannot completely remove these organic drugs, these organic drugs are discharged into the water environment and continuously penetrate into the surface water, the ground water and the sediment, so that the distribution of the organic medicine in the environment is universal. Among them, diclofenac is one of the most commonly used non-opioid analgesic and anti-inflammatory drugs (NSAIDs), which are widely used in clinical treatment. Due to the good curative effect of the diclofenac medicine, the dosage is large, and the diclofenac medicine is discharged to the environment without control, and the stability of the ecological system is endangered. It is now known that diclofenac can severely affect the growth and development of plants, but the mechanism to inhibit plant growth is not yet clear. To this end, the effects of diclofenac on the respiratory metabolism and the mitochondrial function of the tobacco by-2 cells were investigated by using the tobacco BY-2 cells, and the total respiration of the tobacco by-2 cells treated with diclofenac sodium at different concentrations was determined by using the Oxytherm oxygen electrode (Hansatch, UK). the total respiration, the cytochrome pathway (COX) and the alternating oxidase (AOX) pathway of the tobacco by-2 cells and the state respiration of the mitochondria were measured for different treatment times, and the cellular respiration of the different respiratory pathway inhibitor treatment was determined, And the inhibition of the growth of the tobacco by-2 cells by the diclofenac sodium is more visually verified by the method for weighing the dry weight of the cells. In addition, the mechanism of diclofenac to inhibit the growth of the tobacco by-2 cells is explained, and the theoretical basis for elucidating the mechanism of diclofenac to inhibit the growth of the plants is provided. The results of this study show that after 24 h of 0.2 mmol 路 L-1 diclofenac acid treatment, the growth of the tobacco by-2 cells is significantly inhibited, and the occurrence and accumulation of reactive oxygen (ROS) in the cells is induced, and the tobacco by-2 cells are caused to die. In addition, it was found that the concentration of diclofenac has an immediate effect on the respiration of the tobacco by-2 cells, and as the concentration of diclofenac increases, the degree of inhibition of cellular respiration is also increased, and when the concentration of the diclofenac is 0.2 mmol 路 L-1, It has the greatest inhibitory effect on the respiration of the tobacco by-2 cells. In addition, in the study of the direct effect of diclofenac on the in vitro mitochondria of the tobacco by-2 cells, we found that diclofenac has been used to inhibit the activity of the complexes II, III and IV on the mitochondrial respiratory electron transfer chain of the cell, The two respiratory electron transport pathways involved in the participation of the cytochrome oxidase (COX) and the alternating oxidase (AOX) in the mitochondria are inhibited by different degrees, and the glycolysis (EMP) of the cells is further suppressed, The three pathways of carbon metabolism in the three-acid cycle (TCA) and the pentose phosphate pathway (PPP). In addition, diclofenac has an inhibitory effect on the respiration of the three states of the mitochondria I, III and IV, leading to a reduction in the respiratory control rate (RCR). In addition, diclofenac can also lead to the swelling of the mitochondria, and the aggregation of the mitochondrial membrane phospholipids can be reduced, the integrity of the mitochondria is destroyed, the formation of the transmembrane proton gradient is blocked, and the ATP synthesis is inhibited. Therefore, we believe that the inhibition of the electron transfer of complexes III and IV through the mitochondrial respiratory electron transfer chain in the tobacco BY-2 cells is a major cause of diclofenac to inhibit cellular respiration. This study has shown that the inhibition of the activity of diclofenac on the complex II, III and IV of the mitochondrial respiratory chain is the main cause of the inhibition of cellular respiration, which leads to a disorder in the metabolism of the mitochondria. The disorder of mitochondrial energy metabolism and substance metabolism further causes the explosion and accumulation of reactive oxygen (ROS) in the cells, which are important reasons for diclofenac to inhibit the growth of the tobacco by-2 cells and lead to cell death.
【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：X503.231

【共引文獻】

相關期刊論文 前10條

1 皮運清;郭偉;馮精蘭;孫劍輝;;Fenton氧化氧氟沙星過程中間產(chǎn)物的鑒定[J];分析試驗室;2014年04期

2 張娟;馬玉龍;孫瑞珠;王艷;馬琳;;泰樂菌素降解菌的分離及其酶促特性[J];環(huán)境科學與技術;2014年04期

3 徐秋桐;鮑陳燕;章明奎;;土壤中土霉素對12種農(nóng)作物幼苗毒性閾值研究[J];安徽農(nóng)學通報;2014年09期

4 曹貴彬;易鳳珍;游金明;王自蕊;葉亞玲;;陳皮苷制劑對愛拔益加肉雞生長性能、免疫器官指數(shù)及小腸黏膜形態(tài)結構的影響[J];動物營養(yǎng)學報;2014年03期

5 陳海燕;樊霆;葉文玲;魯洪娟;花日茂;;安徽省菜地土壤中3種磺胺類抗生素的殘留調(diào)查[J];安徽農(nóng)業(yè)大學學報;2014年03期

6 郭寧;隋銘?zhàn)?周友飛;盛力;;Al-SBA-15有序介孔材料對溶液中諾氟沙星的吸附研究[J];安全與環(huán)境學報;2014年06期

7 段曉軍;;水中抗生素的處理方法的研究進展[J];廣東化工;2015年11期

8 周林軍;馮潔;劉濟寧;石利利;王濟奎;;污水處理廠中化學品暴露預測模型研究進展[J];環(huán)境科學與技術;2015年08期

9 Shiliang WANG;Hui WANG;;Adsorption behavior of antibiotic in soil environment:a critical review[J];Frontiers of Environmental Science & Engineering;2015年04期

10 李清雪;王釗;祁跡;;萃取/液相色譜同時檢測污水中20種藥物活性物質(zhì)[J];中國給水排水;2015年14期

相關博士學位論文 前10條

1 石義靜;硝化顆粒污泥的培養(yǎng)及其與四環(huán)素相互作用研究[D];山東大學;2013年

2 李志華;卡馬西平對虹鱒的生態(tài)毒理學影響[D];西南大學;2013年

3 雷超;表面活性劑改性分子篩吸附三氯生的性能及機理研究[D];華南理工大學;2013年

4 張德才;SDHB在結直腸癌中的表達及功能研究[D];中南大學;2013年

5 陳楠;非均相類芬頓催化劑用于上流式多相氧化塔處理紅霉素廢水的研究[D];廣西大學;2012年

6 吳維;天津市供水系統(tǒng)中抗生素檢測與控制方法的研究[D];天津大學;2012年

7 徐冰潔;不同碳源條件下功能菌共代謝降解典型PPCPs的效能與機理[D];東華大學;2014年

8 樊慧菊;PAC-MBR工藝去除污水中藥物污染物效能與機制研究[D];北京林業(yè)大學;2014年

9 皮運清;高級氧化技術對典型PPCPs降解效果及降解機理的研究[D];河南師范大學;2013年

10 嚴清;典型PhACs在城市水系統(tǒng)中的遷移分布規(guī)律及其在人工濕地中的去除研究[D];重慶大學;2014年

相關碩士學位論文 前10條

1 閆璐;剩余污泥及其微生物燃料電池處理后抗生素抗性基因水平分析[D];東北農(nóng)業(yè)大學;2013年

2 史文艷;阻斷二氮嗪后處理對大鼠心肌線粒體蛋白質(zhì)表達譜變化的影響[D];遵義醫(yī)學院;2013年

3 王志剛;電化學法對養(yǎng)殖廢水中污染物去除研究[D];西南大學;2013年

4 李曼;再生水灌溉土壤中典型抗生素遷移特征研究[D];河北科技大學;2013年

5 胡秀敏;泰樂菌素與菲/Cd（Ⅱ）雙組分體系在粘土礦物上的吸附特性[D];華南理工大學;2013年

6 衛(wèi)毅梅;抗生素在城市河流中的污染特征及生態(tài)毒性研究[D];遼寧大學;2013年

7 李通;環(huán)丙沙星的生態(tài)毒性及其在高鐵酸鉀作用下的降解研究[D];河南師范大學;2013年

8 楊芳;人工濕地中抗生素耐藥菌和耐藥基因環(huán)境行為研究[D];暨南大學;2013年

9 俞悅;光化學及生物法降解磺胺甲VA唑（SMX）的研究[D];上海師范大學;2013年

10 馬s，

本文編號：2449219