纳米氧化铈对过氧化氢所致大鼠肺泡巨噬细胞氧化损伤的影响

目的研究纳米氧化铈对过氧化氢致大鼠肺泡巨噬细胞NR8383氧化损伤的影响。方法将对数生长期的NR8383细胞分别暴露于终浓度为5、10、20、50、100μg/ml纳米氧化铈(20 nm)悬液孵育24 h,再加入新鲜配制的终浓度为100μmol/L的过氧化氢刺激细胞2 h,另设对照(PBS)组及过氧化氢(100μmol/L)组。采用CCK-8法检测NR8383细胞的细胞活性,采用酶标法检测培养液上清中乳酸脱氢酶(LDH)的释放量,并测定细胞内谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、活性氧(ROS)的水平。结果与对照组相比,过氧化氢对NR8383细胞造成氧化损伤,细胞的存活率下降25.25%。与过氧化氢组相比,5μg/ml纳米氧化铈+过氧化氢组NR8383细胞的存活率较高,而100μg/ml纳米氧化铈+过氧化氢组NR8383细胞的存活率较低,差异均有统计学意义(P0.05,P0.01);且随着纳米氧化铈染毒浓度的升高,NR8383细胞的存活率呈下降的趋势。与过氧化氢组相比,对照组NR8383细胞的LDH释放量较低,50μg/ml纳米氧化铈+过氧化氢组NR8383细胞的LDH释放量较高,差异有统计学意义(P0.05);而5、10μg/ml纳米氧化铈+过氧化氢组NR8383细胞的LDH释放量有所降低,但差异均无统计学意义(P0.05)。且随着纳米氧化铈染毒浓度的升高,NR8383细胞的LDH释放量呈上下波动。与过氧化氢组相比,对照组NR8383细胞内GSH的水平升高,而ROS的水平上升;5、10μg/ml纳米氧化铈+过氧化氢组NR8383细胞内的ROS水平均下降,而10μg/ml纳米氧化铈+过氧化氢组NR8383细胞内的GSH水平上升,差异均有统计学意义(P0.05)。随着纳米氧化铈染毒浓度的升高,NR8383细胞内的SOD、GSH呈上下波动,ROS的水平呈逐渐升高的趋势。结论低浓度(5、10μg/ml)的纳米氧化铈颗粒可以缓解过氧化氢对NR8383细胞造成的氧化损伤,对细胞起到保护作用。

Influence of CeO2 nanoparticles on hydrogen peroxide induced damage in alveolar macrophages cells in rats

Objective To understand the influence of nano cerium oxide on damage induced by hydrogen peroxide in alveolar macrophages cells in rats,to know whether it can alleviate the damage caused by hydrogen peroxide.Methods 20 nm nano cerium oxide of different final concentrations (5,10,20,50 and 100 μg/ml) were applied to NR8383 cells for 24 hours,then 100 μmol/L H2O2 applied to cells for 2 hours.CCK-8 method was used to detect the survival rate of NR8383 cells,enzymelabeled method was used to assay the release of lactate dehydrogenase (LDH) in the culture supernatant,the contents of glutathione (GSH) and superoxide dismutase (SOD).Fluorescence microscopy was used to observe the changes of reactive oxygen species (ROS).Results Compared with the control group,hydrogen peroxide induced oxidative damage to NR8383 cells,the cell survival rate decreased by 25.25％.Compared with hydrogen peroxide group,the viability of NR8383 cells was higher in 5 μg/ml nano cerium oxide+hydrogen peroxide group,while in 100 μg/ml nano cerium oxide+hydrogen peroxide group,the survival rate of NR8383 cells was significantly lower (P＜0.05,P＜0.01),with a dose-dependent manner.Compared with the hydrogen peroxide group,the LDH release of NR8383 cells in the control group was lower,in the 50 μg/ml nano cerium oxide+hydrogen peroxide group,the LDH release of NR8383 cells was significantly higher than that in control group (P＜0.05),while in the 5 μg/ml,10 μg/ml nano-cerium oxide and hydrogen peroxide groups the LDH release of NR8383 cells were decreased,but the difference were not statistically significant (P＞0.05).And with the increase of concentration of nano-cerium oxide,the LDH release fluctuated.Compared with hydrogen peroxide group,the level of GSH in the control group was higher,while the level of ROS increased.In the 5 μg/ml,10 μg/ml nano-cerium oxide and hydrogen peroxide groups,the levels of ROS were decreased.While in the 10 μg/ml nano-cerium oxide and hydrogen peroxide groups,the level of GSH in NR8383 cells was significantly increased (P＜0.05).With the increase of the concentration of nano-cerium oxide,the SOD and GSH in NR8383 cells fluctuated,and the level of ROS increased.Conclusion Low concentration of CeO2 nanoparticles can alleviate the oxidative damage caused by H2O2 on NR8383 cells and protect cells from oxidative damage.