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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(P<0.05,P<0.01);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(P<0.05);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(P>0.05)。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(P<0.05)。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。
Objective To investigate the effect of nano-cerium oxide on oxidative damage of rat alveolar macrophage NR8383 induced by hydrogen peroxide. Methods The logarithmic growth phase of NR8383 cells were exposed to a final concentration of 5,10,20,50,100μg / ml suspension of nano-cerium (20 nm) for 24 h, then added freshly prepared final concentration of 100μmol / L The cells were stimulated by hydrogen peroxide for 2 h. The control group (PBS) and hydrogen peroxide (100 μmol / L) groups were also established. The cell viability of NR8383 cells was detected by CCK-8 assay. The release of lactate dehydrogenase (LDH) in the culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA) and the content of glutathione (GSH), superoxide dismutase (SOD), reactive oxygen species (ROS) levels. Results Compared with the control group, hydrogen peroxide caused oxidative damage to NR8383 cells, and cell viability decreased by 25.25%. Compared with hydrogen peroxide group, the survival rate of NR8383 cells in 5μg / ml nano-ceria + hydrogen peroxide group was higher, while the survival rate of NR8383 cells in 100μg / ml nano-ceria + hydrogen peroxide group was lower (P <0.05, P <0.01). With the increase of concentration of nano-cerium oxide, the survival rate of NR8383 cells decreased. Compared with the hydrogen peroxide group, the LDH release of NR8383 cells in the control group was lower, and the release of LDH was higher in the NR8383 cells treated with 50μg / ml nano-sized ceria + hydrogen peroxide (P <0.05). However, LDH release of NR8383 cells in 5,10μg / ml nano-sized ceria + hydrogen peroxide group decreased, but the differences were not statistically significant (P> 0.05). And with the concentration of nano-cerium increased, the amount of LDH released by NR8383 cells fluctuated. Compared with hydrogen peroxide group, the level of GSH in NR8383 cells increased and the ROS level increased in the control group. The ROS levels in NR8383 cells treated with 5 and 10μg / ml nano-ceria + hydrogen peroxide decreased, while the levels of 10μg / ml nano-ceria + hydrogen peroxide group NR8383 cells increased GSH levels, the differences were statistically significant (P <0.05). With the increase of the concentration of nano-cerium oxide, the intracellular SOD and GSH in NR8383 fluctuated up and down, while the level of ROS increased gradually. Conclusions Nano-sized cerium oxide particles (5, 10μg / ml) can alleviate the oxidative damage caused by hydrogen peroxide to NR8383 cells and play a protective role on the cells.