实时动态监测纳米氧化铜作用于CHL细胞的毒性效应

Real-time dynamic monitoring of cytotoxicity in nano-copper oxide-treated CHL cells

目的：探讨纳米氧化铜连续作用于CHL细胞的毒性效应。方法：应用实时细胞分析仪（RTCA）对不同接种密度（1×105、5×10~4、2.5×10~4、1.25×10~4/mL）的CHL细胞进行140 h连续测定,绘制不同密度CHL细胞的生长曲线。继而对不同剂量（0、2、1、0.5、0.25、0.125、0.062 5、0.031 3 mg/mL）的纳米氧化铜作用于CHL细胞所产生的毒性效应进行实时监测,得到不同作用时间的IC_（50）值,绘制连续时间点IC_（50）变化的曲线,以判定纳米氧化铜作用于CHL细胞所产生的毒性效应。结果：4种不同密度CHL细胞接种后,细胞达到对数生长期的时间有所不同,其中5×10~4/mL密度最适合进行毒性效应试验。不同剂量纳米氧化铜染毒后,毒性作用起始时间为4~5 h,加入受试物后,前6h的IC_（50）值较高,且波动较大。从6h以后,IC_（50）值呈现明显下降,作用12、24、36、48 h的IC_（50）值分别为0.157 0、0.0511、0.0429和0.016 8 mg/mL。结论：使用RTCA实时动态监测系统测定纳米材料的细胞毒性具有实用性。在纳米氧化铜对CHL细胞所产生毒性作用的进一步研究工作中,对染毒后4~18 h所产生的毒性作用进行研究更有意义。

OBJECTIVE：To study the cytotoxicity of CHL cells after treatment with nanometer copper-oxide.METHODS：CHL cells in different densities（1×10~5,5×10~4,2.5×10~4 and 1.25×10~4/mL） were monitored continuously by RTCA and their whole growth curves were evaluated.Then,the cytotoxicity of CHL cells treated by nanometer copper-oxide in different doses（0,2,1,0.5,0.25,0.125,0.062 5 and 0.0313 mg/mL） was real-time monitored,and the accurate IC_（50） value of the continuous time and the curve of IC_（50） in continuous time were studied.RESULTS：Growth curves in cultures with different cell densities were measured,and cultures with 5×10~4/mL inoculated cell number was found the best for toxic effect test.The starting time of the toxic effect after treatment with nano-copper oxide was about 4-5 h.At 6 h after treatment with nanometer copper-oxide,the IC_（50） value was bigger with a large volatility,while it presented a dramatic decline after 6 h.The value of IC_（50） after 12,24,36 and 48 h was 0.157 0,0.0511,0.0429,0.016 8 mg/mL,respectively.CONCLUSION：RTCA was useful for detecting cytotoxicity of nanometer copper-oxide,considering the unique characteristics of nanomaterials.In addition,cytotoxicity at 4-18 h after treatment should be investigated further in the future.