摘要
目的探讨不同粒径纳米氧化锌(Zn O)对不同分化状态人结肠腺癌细胞(Caco-2)的毒性作用。方法将Caco-2细胞接种于96孔板培养24 h(未分化细胞)或21 d(分化细胞),选用3种尺寸纳米氧化锌,分别为Zn O-20(25.3±5.8 nm)、Zn O-L(25.6±5.7 nm)和Zn O-100(96.7±42.7 nm),设置不同的剂量(6.25、12.5、25、50和100μg/ml)与未分化或分化的Caco-2细胞共培养24、48或72 h后,采用CCK-8法检测细胞活性,比色法检测乳酸脱氢酶(LDH)释放率以及碱性磷酸酶(ALP)活性。结果浓度为100μg/ml处理72 h,3种纳米氧化锌可使未分化细胞活性分别下降81.3%、68.1%和55.1%,分化细胞活性分别下降69.3%、55.1%和49.0%,染毒24 h,在50μg/ml的浓度条件下LDH释放率分别升高至70.6%、68.4%和54.7%,影响细胞膜结构的完整性。并且对ALP表达均有抑制作用,上述作用均存在剂量反应关系。纳米氧化锌粒径越小毒性越大,毒性强弱顺序为Zn O-20>Zn O-L>Zn O-100,粒径相近时,毒性仍有差别,可能与颗粒的形状等其他性质有关。此外,对于同一种纳米材料毒性作用,未分化细胞的敏感性较分化细胞强。结论纳米氧化锌对Caco-2细胞的毒性作用与其粒径、形状等理化性质以及细胞分化状态均相关。
Objective To investigate the toxic effects of zinc oxide(Zn O） nanoparticles with different particle sizes in human colon adenocarcinoma cells(Caco-2） in different differentiation status. Methods Caco-2 cells were seeded in 96-well plates for 24 h(undifferentiated cells） or 21 days(differentiated cells）,three types of Zn O nanoparticles were selected and used as Zn O-20(25. 3 ± 5. 8 nm）,Zn O-L(25. 6 ± 5. 7 nm） and Zn O-100(96. 7 ± 42. 7 nm）,Zn O nanoparticles were co-cultured with undifferentiated or differentiated Caco-2 cells for 24,48 or 72 h at doses of 6. 25,12. 5,25,50 and 100 μg/ml. The cell viability was measured by CCK-8 assay. The release rate of lactate dehydrogenase(LDH） and the activity of alkaline phosphatase(ALP） were determined by colorimetric assay. Results The three kinds of nano-Zn O could decrease the activity of undifferentiated cells by 81. 3%,68. 1% and 55. 1%,respectively,the activities of differentiated cells by 69. 3%,55. 1% and 49. 0%,respectively at the concentration of 100 μg/ml co-cultured for 72 h. The LDH release rate increased to 70. 6%,68. 4%,54. 7% respectively at the concentration of 50 μg/ml for 24 h. The membrame intergrity was damaged,and the ALP expression was inhibited. The above changes were dose-response related. Under the same concentration conditions,the smaller the particle size,the greater the toxicity,and the order of the toxicity is Zn O-20 Zn O-L Zn O-100. Toxicity differences between Zn O-20 and Zn O-L indicate that the toxicity may also depend on the particle shape and other properties. In addition,Zn O nanoparticles are more toxic to undifferentiated cells than to differentiated cells. Conclusion The toxic effects of Zn O nanoparticles in Caco-2 cells are related to physicochemical properties of nanoparticles and cell differentiation status.
作者
熊雰
梁春来
于洲
徐海滨
XIONG Fen;LIANG Chun-lai;YU Zhou;XU Hai-bin(Ningxia Medical University, Yinchuan, Ningxia 750004, China;Key Laboratory of Food Safety Risk Assessment of Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China)
出处
《毒理学杂志》
CAS
CSCD
2018年第2期87-92,共6页
Journal of Toxicology
基金
国家食品安全风险评估中心高层次人才队伍建设523项目(CFSA“523”)
