Nowrouzi Azin, Meghrazi Khadijeh, Golmohammadi Taghi, Golestani Abolfazl, Ahmadian Shahin, Shafiezadeh Mahshid, Shajary Zahra, Khaghani Shahnaz, Amiri Azita N
Dept. of Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Iran Biomed J. 2010 Jan-Apr;14(1-2):23-32.
We aimed at evaluating the toxicity effects of low (subtoxic) concentrations of silver nanoparticles nanoparticles (AgNP, 5-10 nm) in human hepatoblastoma (HepG2) cell line after and during a period of about one month.
XTT and MTT assays were used to draw a dose-response curve; IC50 (half maximal inhibitory concentration) value of the AgNP on HepG2 cells was calculated to be 2.75-3.0 mg/l. The cells were exposed to concentrations of 0% (control), 1%, 4% and 8% IC50 of AgNP (corresponding to 0.00, 0.03, 0.12 and 0.24 mg/l of AgNP, respectively) for four consecutive passages. The treated cells were compared to the control group with respect to morphology and proliferation at the end of the period.
The biochemical studies revealed significant increases of lactate dehydrogenase and alanine aminotransferase enzyme activity in the culture media of cells receiving 4% and 8% IC50; the increases in the aspartate aminotransferase enzyme activity and nitric oxide concentration became significant at 8% IC50. In the cell extracts, the average total protein and activity of glutathione peroxidase enzyme remained unchanged; the decrease in the average content of glutathione (GSH) and superoxide dismutase (SOD) activity became significant at 4% and 8% IC50. There were increases in lipid peroxidation (significant at 4% and 8% IC50) and cytochrome c content (significant at 8% IC50). The accumulations of the effects, during the experiment from one generation to the next, were not statistically remarkable except in cases of GSH and SOD. The results indicate clearly the involvement of oxidative changes in the cells after exposure to low doses of AgNP.
The results might help specify a safer amount of AgNP for use in different applications.
我们旨在评估低(亚毒性)浓度的银纳米颗粒(AgNP,5 - 10纳米)在约一个月期间及之后对人肝癌细胞系(HepG2)的毒性作用。
采用XTT和MTT试验绘制剂量反应曲线;计算出AgNP对HepG2细胞的IC50(半最大抑制浓度)值为2.75 - 3.0毫克/升。细胞连续传代四次,分别暴露于0%(对照)、1%、4%和8% IC50浓度的AgNP(分别对应0.00、0.03、0.12和0.24毫克/升的AgNP)。在该时间段结束时,将处理后的细胞与对照组在形态和增殖方面进行比较。
生化研究显示,接受4%和8% IC50浓度AgNP处理的细胞培养基中乳酸脱氢酶和丙氨酸转氨酶活性显著增加;天冬氨酸转氨酶活性和一氧化氮浓度在8% IC50时显著增加。在细胞提取物中,谷胱甘肽过氧化物酶的平均总蛋白和活性保持不变;谷胱甘肽(GSH)平均含量和超氧化物歧化酶(SOD)活性在4%和8% IC50时显著降低。脂质过氧化增加(在4%和8% IC50时显著),细胞色素c含量增加(在8% IC50时显著)。除GSH和SOD外,实验过程中从一代到下一代的效应累积在统计学上不显著。结果清楚地表明,低剂量AgNP暴露后细胞中发生了氧化变化。
这些结果可能有助于确定在不同应用中使用AgNP的更安全剂量。