氧氟沙星和铜及其络合体系对非洲绿猴肾细胞（Vero）的毒性效应

发布时间：2018-06-16 19:50

  本文选题：氧氟沙星 + 重金属铜　； 参考：《昆明理工大学》2017年硕士论文

【摘要】：抗生素的滥用使其通过多种途径进入环境,从而产生健康风险。重金属在土壤及水体环境中普遍存在,其隐蔽性强的特性导致它容易通过食物链富集,造成动植物出现重金属中毒。抗生素及重金属在环境中的共存现象非常普遍,并且抗生素易与重金属离子发生络合反应,使它们的环境风险评估变得更加复杂。研究抗生素与重金属对环境中生物的复合风险势在必行。本研究选择抗生素中的代表性物质氧氟沙星(OFL)和重金属中的铜离子(Cu~(2+))作为实验对象,对离体细胞培养中的模式细胞非洲绿猴肾细胞(Vero)进行复合污染物毒性实验,以此研究抗生素OFL与Cu~(2+)及其复合污染物对Vero细胞的生物效应。通过对比OFL、Cu~(2+)及其复合污染物对Vero细胞的毒性效应的差异研究复合污染与单独污染的区别,并进一步探讨影响氧氟沙星和Cu~(2+)络合物形成的影响因素(pH值和温度)及复合络合体系对Vero细胞的毒性效应。所以本研究对抗生素与重金属复合污染的环境风险研究具有一定理论和实践价值。结果表明:(1)OFL与Cu~(2+)及其复合污染物会导致细胞形态变化,细胞破碎,贴壁率降低。当Cu~(2+)浓度从6.88 mg·L-1增加到6875 mg·L-1时,其对Vero细胞生长的抑制率也上升了 55%。实验表明,OFL对Vero细胞的生长也有显著的影响,当OFL浓度低于3mg·L-1时,OFL促进了 Vero细胞的生长;而当OFL浓度高于3 mg·L-1时,Vero细胞的生长则被抑制。(2)氧氟沙星和铜复合污染对Vero细胞的毒性与其污染物单独污染时的毒性之和相比有所下降。在2.75 mg·L-1Cu~(2+)浓度时,OFL浓度从0.63 mg·L-1增加到10 mg· L-1时其复合污染物对细胞的抑制率从24%上升到了最高47%。当OFL浓度为 2.5 mg·L-1、5 mg·L-1 和 10 mg·L-1 时,OFL 与浓度为 2.75 mg·L-1 Cu~(2+)的复合物对细胞的抑制率与OFL、Cu~(2+)单一抑制率之和有显著差异,呈现出抑制率下降的趋势。同时,当Cu~(2+)浓度6.88mg·L-1处理下时出现了类似的现象,OFL与浓度为6.88 mg· L-1 Cu~(2+)的复合物对细胞抑制率显著低于OFL、Cu~(2+)单一抑制率之和。(3)DMEM培养基中OFL与Cu~(2+)的游离的量要显著低于水环境中两者游离的量。说明OFL和Cu~(2+)会与DMEM培养基中其他物质发生反应,形成一个由OFL与Cu~(2+)为主的复合络合体系。(4)温度和pH值的变化能够对DMEM细胞培养液中的OFL-Cu~(2+)复合络合体系总浓度产生显著的影响。随着温度的升高,OFL-Cu~(2+)复合络合体系总浓度逐渐增大,这说明温度的升高不但增大了氧氟沙星和铜离子之间的相互作用,还提高了它们与DMEM培养液中其他物质的反应活性。同时,pH值的增大也使得OFL-Cu~(2+)复合络合体系的总浓度上升,这说明pH值的增大能够提高氧氟沙星与铜的反应活性及其络合能力。(5)OFL-Cu~(2+)复合络合体系不会对Vero细胞的生长产生毒性效应。OFL和Cu~(2+)复合污染毒性与单独污染毒性之和相比有所下降的主要原因之一是在氧氟沙星与铜复合污染体系中DMEM细胞培养液、OFL和Cu~(2+)相互之间发生了反应,形成了 OFL-Cu~(2+)复合络合体系,而该体系对Vero细胞的生长产生的影响不显著,从而降低了氧氟沙星和铜复合污染的总体毒性。由此看来,抗生素OFL与重金属Cu~(2+)对Vero细胞的复合毒性会因为OFL-Cu~(2+)复合络合体系的产生而下降。说明OFL与Cu~(2+)对Vero细胞生长的毒性效应具有拮抗作用。表明环境中污染物间可能存在拮抗作用使复合污染对环境的风险下降。
[Abstract]:The abuse of antibiotics causes it to enter the environment through a variety of ways, resulting in health risks. Heavy metals are prevalent in the soil and water environment. Their concealed and strong characteristics cause it to be easily enriched through the food chain, causing heavy metal poisoning in animals and plants. The coexistence of antibiotics and heavy metals in the environment is very common and resistant to the environment. In this study, the representative substances ofofloxacin (OFL) and copper ions (Cu~ (2+)) in heavy metals (Cu~ (2+)) are selected as the experimental object, and in vitro finer. The cytotoxic effects of antibiotic OFL and Cu~ (2+) and their compound pollutants on Vero cells were studied in the cytotoxicity test of cell culture model cells, African green monkey kidney cells (Vero), and the difference between compound pollution and single pollution was studied by comparing the differences of OFL, Cu~ (2+) and its compound pollutants to Vero cells. The influence factors of the formation of ofloxacin and Cu~ (2+) complex formation (pH value and temperature) and the toxic effect of complex complex system on Vero cells are further explored. Therefore, this study has a certain theoretical and practical value on the environmental risk study of the compound pollution of antibiotics and heavy metals. The results show that: (1) OFL and Cu~ (2+) and its compound pollutants will be found. When Cu~ (2+) concentration increased from 6.88 mg. L-1 to 6875 mg. L-1, the inhibition rate to the growth of Vero cells was also increased by 55%. experiment, which showed that OFL had a significant influence on the growth of Vero cells, and when OFL concentration was lower than 3mg. The growth of Vero cells was inhibited when higher than 3 mg. L-1. (2) the toxicity of the compound pollution of ofloxacin and copper to Vero cells was decreased compared with the toxicity of the contaminants alone. At the concentration of 2.75 mg L-1Cu~ (2+), the concentration of OFL from 0.63 mg L-1 increased to 10 mg L-1. The inhibitory rate of the compound pollutants to the cells was 24%. When the concentration of OFL was 2.5 mg. L-1,5 mg, L-1 and 10 mg. L-1, the inhibition rate of OFL and concentration of 2.75 mg. L-1 Cu~ was significantly different from that of the single inhibitory rate. The cell inhibition rate of OFL and the concentration of 6.88 mg. L-1 Cu~ (2+) was significantly lower than that of OFL, Cu~ (2+) single inhibition rate. (3) the free amount of OFL and Cu~ (2+) in the DMEM medium was significantly lower than that in the water environment. 2+) complex complex system. (4) the change of temperature and pH value can significantly influence the total concentration of OFL-Cu~ (2+) complex complex system in DMEM cell culture. With the increase of temperature, the total concentration of OFL-Cu~ (2+) complex complex system increases gradually. This shows that the increase of temperature not only increases the phase between ofloxacin and copper ions. The interaction between them and other substances in the DMEM culture also increased their reactivity to other substances in the DMEM culture. At the same time, the increase of the pH value also increased the total concentration of the OFL-Cu~ (2+) complex complex system, which indicated that the increase of the pH value could improve the reactivity and complexation ability of ofloxacin and copper. (5) the OFL-Cu~ (2+) complex complex system would not produce Vero cells. One of the main reasons for the decrease of the toxic effect of.OFL and Cu~ (2+) on the toxicity of the compound pollution toxicity of the DMEM cell in the compound pollution system of ofloxacin and copper is the reaction between OFL and Cu~ (2+) in the compound pollution system of ofloxacin and copper, and the complex complex system of OFL-Cu~ (2+) is formed, and the growth of Vero cells in this system The effect of the production is not significant, thus reducing the overall toxicity of the compound pollution of ofloxacin and copper. Therefore, the compound toxicity of antibiotic OFL and heavy metal Cu~ (2+) to Vero cells will be reduced by the production of OFL-Cu~ (2+) complex complex system. It shows that the toxic effects of OFL and Cu~ (2+) on the growth of Vero cells are antagonistic. There may be antagonism among pollutants, which makes the risk of compound pollution to environment decrease.
【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X171.5

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