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目的研究汉阳地区PM2.5对人胚胎干细胞来源的成纤维细胞(EBf-H9细胞)的遗传毒性,验证EBf-H9细胞作为遗传毒性评价模型的可行性,拟为遗传毒性评价提供一个新的可选细胞模型。方法不同浓度PM2.5作用于EBfH9细胞24 h,应用CCK-8试剂盒及LDH试剂盒检测EBf-H9细胞的细胞毒性和LDH漏出率;采用单细胞凝胶电泳试验及微核试验评价PM2.5对EBf-H9细胞DNA和染色体损伤程度。结果细胞毒性试验结果显示,随着PM2.5剂量的增加,细胞存活率逐渐降低,而LDH漏出率呈先增加后降低的趋势,且存在剂量-效应关系;单细胞凝胶电泳试验与双核细胞微核试验结果显示,随着PM2.5剂量的增加,彗星拖尾率与双核细胞微核率逐渐上升,且存在明显的剂量-效应关系。结论在本试验条件下,汉阳PM2.5引起EBf-H9细胞出现DNA和染色体损伤,初步证实EBf-H9细胞可作为一种新的遗传毒性评价候选细胞模型。
Objective To investigate the genetic toxicity of PM2.5 to human embryonic stem cells (EBf-H9) cells in Hanyang area and to verify the feasibility of using EBf-H9 cells as a model for evaluating genotoxicity. Cell model selection. Methods PMF2 cells were treated with different concentrations of PM2.5 for 24 h. Cytotoxicity and LDH leakage rate of EBf-H9 cells were detected by CCK-8 kit and LDH kit. PM2 was evaluated by single cell gel electrophoresis and micronucleus test. 5 on EBf-H9 cell DNA and chromosome damage. Results The results of cytotoxicity showed that with the increase of PM2.5 dosage, the cell viability decreased and the leakage rate of LDH first increased and then decreased, and the dose-response relationship existed. The results of single cell gel electrophoresis The results of micronucleus test showed that with the increase of PM2.5 dose, the tailing rate of comets and micronuclei of binuclear cells gradually increased, and there was a significant dose-response relationship. Conclusion Under the experimental conditions, DNA and chromosomal damage of EBf-H9 cells induced by Hanyang PM2.5 may be preliminary evidence that EBf-H9 cells can be used as a new candidate model of genotoxicity evaluation.