硫酸铜和氯化锌对不同倍性泥鳅鳍细胞系的毒性效应

发布时间：2017-12-27 01:31

本文关键词：硫酸铜和氯化锌对不同倍性泥鳅鳍细胞系的毒性效应　出处：《大连海洋大学》2015年硕士论文　论文类型：学位论文

【摘要】：本文研究了不同浓度的硫酸铜和氯化锌对70-80代的二倍体泥鳅鳍细胞系(DIMF)和三倍体泥鳅鳍细胞系(TRMF)的毒性效应。采用四噻唑蓝(MTT)比色法测得DIMF和TRMF的硫酸铜的半致死浓度分别为154.3士23μmol/L、184.6士14μmol/L。对氯化锌的半致死浓度为245.2士12μmol/L、293.6士23μmol/L。三倍体泥鳅鳍细胞系半致死浓度明显高于二倍体泥鳅鳍细胞系(P0.05)。DIMF和TRMF中酶活性的测定结果显示:经硫酸铜处理后,二倍体泥鳅鳍细胞系中超氧化物歧化酶(SOD)的活性在硫酸铜浓度为0~200μmol/L范围Qg,随浓度的升高逐渐升高,在200μmol/L浓度时达到最大值,随后逐渐降低。三倍体泥鳅鳍细胞系中SOD的活性,随硫酸铜浓度的增加逐渐降低。谷胱甘肽-S-转移酶(GST)和谷胱甘肽过氧化物酶(GSH-Px)的活性在两种细胞系中随着硫酸铜浓度的增加呈现逐渐下降的趋势。TRMF细胞中三种酶的活性均高于DIFM。经氯化锌处理后,以上三种酶活性均出现先升高再降低的趋势,当氯化锌浓度为100μmol/L时酶活性达到最大值。微核试验检测硫酸铜对细胞核的损伤,发现DIMF和TRMF细胞中均有微核产生,且微核率随硫酸铜浓度增加逐渐提高。在浓度为400μmol/L时达到最大值,二倍体为3.33‰,三倍体为4.33‰。经氯化锌处理后,氯化锌浓度为0~500μmol/L时,微核率随浓度增加而增加。浓度到达500μmol/L时,二倍体和三倍体微核率均达到最大值,二倍体为3.33‰,三倍体是4.33‰,以后随浓度增加没有显著变化(P0.1)。但在相同浓度下三倍体的微核率要高于二倍体的微核率(P0.1)。超微结构观察结果:对照组DIMF和TRMF中细胞器没有明显差异,但TRMF囊泡较多。经过硫酸铜处理后,DIMF和TRMF细胞的病理变化情况相似,表现为细胞界限模糊,细胞核膜结构不完整,线粒体嵴解体,粗面内质网膜溶解,核糖体脱落游离于胞质中,表明细胞出现了坏死。经过氯化锌处理的细胞,二倍体与三倍体的病理变化较一致:核解体、线粒体水肿、溶酶体形成吞噬小泡、胞浆浓缩颜色较深。表现出细胞凋亡的特征。
[Abstract]:The toxic effects of copper sulfate and zinc chloride on 70-80 generation diploid loach fin cell line (DIMF) and triploid loach fin cell line (TRMF) were studied. The semi lethal concentration of copper sulfate with four thiazolium (MTT) colorimetric method for measuring DIMF and TRMF was 154.3, 23, and 184.6, 14, mol/L, respectively. The semi lethal concentration of zinc chloride was 245.2, 12 mol/L, 293.6 and 23 mu mol/L. The semi lethal concentration of the fin cell line of triploid loach was significantly higher than that of the diploid loach fin cell line (P0.05). The determination results of enzyme activity of DIMF and TRMF in the display: after copper sulfate treatment, diploid loach fin cells of superoxide dismutase (SOD) activity in the copper sulfate concentration was 0~200 mol/L Qg, gradually increased with the concentration increased, reached the maximum at 200 mol/L concentration, then decreased gradually. The activity of SOD in the fin cell line of triploid loach gradually decreased with the increase of copper sulfate concentration. The activity of glutathione -S- transferase (GST) and glutathione peroxidase (GSH-Px) in two cell lines showed a decreasing trend with the increase of cupric sulfate concentration. The activity of three enzymes in TRMF cells was higher than that of DIFM. After the treatment of zinc chloride, the activity of the above three enzymes increased first and then decreased. When the concentration of zinc chloride was 100 mol/L, the enzyme activity reached the maximum. Micronucleus test was used to detect the damage of copper sulfate to the nucleus. Micronuclei were found in DIMF and TRMF cells, and the micronucleus rate increased gradually with the increase of copper sulfate concentration. When the concentration is 400 mol/L, the maximum is reached, the diploid is 3.33 per 1000, and the triploid is 4.33 per thousand. After zinc chloride treatment, the micronucleus rate increases with the concentration of 0~500 micronucleus when the concentration of zinc chloride is mol/L. When the concentration reaches 500 micron mol/L, the rate of micronucleus of diploid and triploid reaches the maximum value, diploid is 3.33 per thousand, triploid is 4.33 per thousand, and it has no significant change with increasing concentration (P0.1). However, the micronucleus rate of triploid in the same concentration is higher than the micronucleus rate of diploid (P0.1). The results of ultrastructure observation: there was no significant difference in the organelles between DIMF and TRMF in the control group, but there were more TRMF vesicles. After the treatment of copper sulfate, the pathological changes of DIMF and TRMF cells were similar. The cells showed fuzzy boundaries, incomplete nuclear membrane structure, disintegration of mitochondria cristae, dissolution of rough endoplasmic reticulum, ribosome exfoliation and dissociation in cytoplasm, indicating necrosis of cells. After zinc chloride treatment, the pathological changes of diploid and triploid were consistent: nuclear disintegration, mitochondrial edema, lysosome formation, phagocytosis, and cytoplasm thickening. It shows the characteristics of cell apoptosis.
【学位授予单位】：大连海洋大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X171.5

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