钼镉联合染毒对鸭肾小管上皮细胞的氧化损伤作用

发布时间：2018-05-08 22:38

本文选题：钼 + 镉　；参考：《江西农业大学》2017年硕士论文

【摘要】：为研究钼镉联合诱导对鸭肾小管上皮细胞氧化损伤的影响。本实验通过筛网过滤法和酶消化法建立鸭肾小管上皮细胞体外培养模型,(NH4)6Mo7O24·4H2O和3CdSO4·8H2O分别作为钼源和镉源,用碱性磷酸酶化学染色(改良钙钴法)、MTT法、冠式法分别对细胞进行鉴定、测定肾小管上皮细胞活性和钼镉单独染毒12 h的IC50。对传一代肾小管上皮细胞进行钼(480μmol/LMo、960μmol/LMo)、镉(2.5μmol/LCd、5μmol/LCd)及其联合(480μmol/L Mo+2.5μmol/L Cd、960μmol/L Mo+5μmol/L Cd)染毒,测定染毒3 h、6 h、12 h、24 h细胞存活率、染毒12 h LDH释放率和SOD活性、T-AOC能力和GSH、MDA含量;并进一步测定NAC对染毒后细胞存活率和LDH释放率的影响。结果表明:(1)碱性磷酸酶化学染色显示,阳性细胞被染为黑色而阴性对照细胞不被染色,对阳性细胞进行计数,显示近端肾小管上皮细胞纯度为95%以上。(2)原代细胞生长曲线:24 h前处于生长期,24 h~88 h处于对数期且78h细胞活性最强,90 h进入平稳期。传一代细胞生长曲线:12 h前处于生长期,12 h~70 h处于对数期且56 h细胞活性最强,70 h进入平稳期。(3)(NH4)6Mo7O24·4H2O和3CdSO4·8H2O染毒12 h的IC50分别为1773.64μmol/LMo、24.17μmol/LCd。(4)细胞存活率:高剂量钼组、镉组及其联合组从3 h开始,极显著低于对照组(P0.01);低剂量钼组从6 h开始,显著低于对照组(P0.05),且呈剂量-时间效应;同一染毒时间,各联合染毒组的细胞存活率均低于各相关单独染毒组;析因分析表明,从染毒12 h开始钼镉之间存在交互作用(P0.05)。NAC可以提高染毒组细胞的存活率(P0.01)。(5)细胞内LDH释放率:各染毒组与对照组相比极显著升高(P0.01),且钼镉联合染毒组高于单独染毒组,NAC对降低LDH释放率无明显作用(P0.05)。(6)SOD活性、T-AOC能力及GSH含量:染毒组均显著或极显著低于对照组(P0.05或P0.01);MDA含量:染毒组均极显著高于对照组(P0.01)。结论:钼镉及其联合染毒对鸭肾小管上皮细胞的氧化损伤呈剂量-时间效应,钼镉呈协同效应;NAC对钼镉及其联合所致的肾小管上皮细胞毒性损伤有一定的保护作用。
[Abstract]:To study the effect of molybdenum and cadmium combined with cadmium on oxidative damage of duck renal tubular epithelial cells. In this experiment, the model of duck renal tubular epithelial cell culture in vitro was established by screening filter method and enzyme digestion method. 4H2O and 3CdSO4 8H2O were used as molybdenum source and cadmium source respectively. The cells were identified by alkaline phosphatase staining (modified Calcium-Cobalt method, Coronal method, respectively). The activity of renal tubular epithelial cells and IC50 induced by molybdenum and cadmium alone for 12 h were measured. A new generation of renal tubular epithelial cells was treated with Mo-480 渭 mol / L Mo-960 渭 mol 路L ~ (-1), CD ~ (2. 5) 渭 mol / L ~ (5 渭 mol 路L ~ (L) and combined with 480 渭 mol/L Mo ~ (2.5 渭 mol/L) CD ~ (6) mol/L Mo ~ (9) 渭 mol/L Mo ~ (5 渭 mol/L). The cell survival rate, the LDH release rate and the activity of SOD, T-AOC and GSH-MDA content were measured at 3 h, 6 h, 12 h and 24 h after exposure. The effects of NAC on cell survival rate and LDH release rate were further determined. The results showed that the positive cells were stained black but the negative control cells were not stained, and the positive cells were counted. The results showed that the purity of proximal tubular epithelial cells was more than 95%.) the growth curve of primary cells was in the growth phase at 24 h and in logarithmic phase at 88 h before 0. 24 h, and the highest cell activity at 78 h was in stable phase at 90 h. The growth curve of the first generation of cells was in the growth phase for 12 h and in the logarithmic phase for 70 h, and the cell activity of 56 h was the highest in the stationary phase. The IC50 of NH4Mo6Mo7O24 4H2O and 3CdSO4 8H2O exposed to 3CdSO4 8H2O for 12 h were 1773.64 渭 mol / L Mo-24.17 渭 mol / L LCd.m4) cell survival rate: high dose molybdenum group, the survival rate of the cells was 1773.64 渭 mol / L Moo 24.17 渭 mol / L LCd.O4, respectively. The cadmium group and its combined group were significantly lower than the control group (P 0.01) from 3 h, and the low dose molybdenum group from 6 h, significantly lower than the control group (P 0.05), and showed a dose-time effect. The cell survival rate of each combined exposure group was lower than that of each other. The interaction between molybdenum and cadmium (P0.05N. NAC) could increase the survival rate of cells and the release rate of LDH in the cells of the exposed group was significantly higher than that of the control group, and the combination of molybdenum and cadmium group was higher than that of the control group. There was no significant effect of NAC on reducing the release rate of LDH. The activity of P0.05N. 6C and the activity of T-AOC and the content of GSH were significantly or extremely lower in the exposed group than those in the control group (P0.05 or P0.01). The content of LDH in the exposed group was significantly higher than that in the control group (P 0.01). Conclusion: the oxidative damage of duckling renal tubular epithelial cells induced by molybdenum and cadmium combined with NAC has a dose-time effect and a synergistic effect of molybdenum and cadmium. NAC has a certain protective effect on the toxicity of renal tubular epithelial cells induced by molybdenum and cadmium.
【学位授予单位】：江西农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S859.8

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