单壁碳纳米管对血管外膜成纤维细胞的毒性作用

CYTOTOXICITY OF SINGLE WALL CARBON NANOTUBES ON RAT VASCULAR ADVENTITIAL FIBROBLASTS

目的探讨不同剂量单壁碳纳米管(SWCNTs)对原代培养大鼠血管外膜成纤维细胞(VAF)的毒性作用,为SWCNTs的毒理学及生物安全性研究提供参考。方法将SWCNTs制成颗粒悬液,设9个剂量组(0.80、1.60、3.20、6.25、12.50、25.00、50.00、100.00、200.00μg/ml)对原代培养Wistar大鼠VAF细胞进行24 h染毒,另设对照组。利用噻唑蓝(MTT)比色法和四唑单钠盐法(WST-1)检测SWCNTs对VAF细胞活性的影响。在此基础上计算出半致死浓度(IC50),再设5个剂量组(0.80、3.20、12.50、50.00、200.00μg/ml),另设对照组,检测SOD活性和MDA含量,以评价细胞的氧化损伤程度。结果 MTT实验结果显示,当剂量为100.00μg/ml时,细胞存活率为41.0%,明显低于对照组(72.3%),(P<0.05);WST-1实验显示,当剂量为50.00μg/ml时,细胞存活率为51.2%,明显低于对照组(77.2%),(P<0.05)。当剂量为12.50μg/ml时,SOD活性为9.61 U/mg prot,明显低于对照组(14.24 U/mg prot),(P<0.05)。当剂量为3.20μg/ml时,MDA含量为0.30 nmol/mg prot,明显高于对照组(0.23 nmol/mg prot),(P<0.05)。结论一定剂量的SWCNTs悬液可抑制VAF细胞活性,导致细胞氧化应激,产生毒性损伤。

Objective To explore the toxic effect of single wall carbon nanotubes（SWCNTs） at different doses on vascular adventitial fibroblasts（VAF）,and provide the basis for toxicological and biological safety study of SWCNTs.Methods Primary VAF cells were exposed to SWCNTs suspension at nine different concentrations（0.80,1.60,3.20,6.25,12.50,25.00,50.00,100.00,200.00 μg/ml） for 24 h.MTT and WST-1 assay were used to evaluate the cell viability.Then VAF cells were exposed to SWCNTs suspension for 48 h at five different concentrations（0.80,3.20,12.50,50.00,200.00 μg/ml） based on the IC50 found.MDA and SOD were used to test the degree of oxidative damage.Results MTT showed that the cell viability was 41.0% at a dose of 100 μg/ml,and different from that of control（72.3%）,（P0.05）;WST showed that the cell viability was 51.2% at a dose of 50 μg/ml,different from that of control（77.2%）,（P0.05）.The activity of SOD（9.61 U/mg prot） at a dose of 12.50 μg/ml was lower than that of control（14.24 U/mg prot）,（P0.05）.The content of MDA（0.30 nmol/mg prot） at 3.20 μg/ml was also lower than that of control（0.23 nmol/mg prot）,（P0.05）.Conclusion SWCNTs suspension of certain concentration could decrease VAF cell viability,and induce oxidative stress,thus result in toxicity to the cell.