香烟导致A549细胞遗传物质损伤的实验研究

发布时间：2018-03-28 19:13

本文选题：香烟烟雾提取物CSE　切入点：彗星实验　出处：《武汉科技大学》2012年硕士论文

【摘要】：目的香烟燃烧产物中含有多种致突变物和致癌物，可以对人体产生严重的遗传损伤，本实验通过提取香烟燃烧产物制作香烟烟雾提取物，并用它刺激肺泡上皮细胞，观察香烟对肺泡上皮细胞的DNA损伤，初步探讨香烟导致肺泡上皮细胞遗传损伤。方法培养人肺泡上皮细胞A549，以二甲基亚砜(DMSO)作为溶剂制取香烟烟气提取物(cigarette smoke extract,CSE)，将不同浓度的CSE作用于A549细胞，用MTT法测定细胞活性，检测DNA-蛋白质交联率（DNA-protein crosslinks,DPC)及彗星实验观察DNA损伤，同时检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶（GSH-PX）、丙二醛（MDA）观察CSE对肺泡细胞的氧化损伤作用。结果 1.10%、20%、50%CSE作用组与对照组相比细胞活性无明显差异，，（P㧐0.05）活性无明显影响，而100%CSE组的细胞活性则明显下降且差异有统计学意义（P㩳0.05）； 2.CSE各浓度组与空白对照组及溶剂组相比，尾部DNA含量，尾长，尾距，Olive尾距均明显增高，差异有统计学意义（P㩳0.05），20%CSE及以上浓度组均表现DPC增高，两指标均有剂量效应关系。 3.20%CSE及以上剂量组的SOD、GSH-PX与对照组相比明显下降， MDA则上升，（P㩳0.05），有一定的剂量效应关系。 4.DPC与抗氧化指标之间存在相关性（P㩳0.05）。结论 1.CSE可导致A549细胞内氧化损伤增加。 2. CSE可导致A549细胞DNA损伤。 3. CSE可导致A549细胞内氧化损伤指标和其导致的DPC率存在相关性。
[Abstract]:Purpose. Cigarette burning products contain many mutagens and carcinogens, which can cause serious genetic damage to human body. In this experiment, cigarette smoke extracts were extracted from cigarette burning products and stimulated alveolar epithelial cells. To observe the DNA damage of alveolar epithelial cells induced by cigarette, and to explore the genetic damage of alveolar epithelial cells induced by cigarette. Method. Human alveolar epithelial cells (A549) were cultured with dimethyl sulfoxide (DMSO) as solvent to prepare cigarette smoke extract smoke extract CSE. Different concentrations of CSE were treated on A549 cells. The cell activity was determined by MTT assay. DNA-protein crosslinking rate (DNA-protein crosslinking DPCs) and comet assay were used to observe the damage of DNA, superoxide dismutase (SOD), glutathione peroxidase (GSH-PXX) and malondialdehyde (MDA) to observe the oxidative damage of CSE to alveolar cells. Results. There was no significant difference in cell activity between CSE group and control group. The cell activity of 100%CSE group was significantly decreased and the difference was statistically significant. 0.05; Compared with the control group and solvent group, the DNA content, tail length, tail distance and Olive tail distance of 2.CSE group were significantly higher than those of control group and solvent group, and the difference was statistically significant. The concentration of DPC was increased in the group of 0. 05 and 20% CSE, both of which had dose-effect relationship. Compared with the control group, the GSH-PX of 3.20%CSE and above dose group decreased significantly, while MDA increased. There is a dose-effect relationship between the two groups. There is a correlation between 4.DPC and antioxidant index. 0.05. Conclusion. 1.CSE induced increased oxidative damage in A549 cells. 2. CSE induced DNA damage in A549 cells. 3. There was a correlation between the index of oxidative damage and the rate of DPC induced by CSE in A549 cells.
【学位授予单位】：武汉科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R114

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