纳米二氧化硅对体外培养A549细胞氧化应激与凋亡的影响

发布时间：2019-05-10 21:49

【摘要】：[目的]研究纳米二氧化硅染毒对人肺腺癌A549细胞氧化应激与凋亡的影响。[方法]用质量浓度(后称"浓度")为25、50、100、200 mg/L的纳米二氧化硅溶液染毒A549细胞,染毒时长为24、48 h,对照组不予处理。用荧光显微镜观察细胞形态变化;采用细胞计数试剂盒(cell counting kit,CCK-8)检测各组在各时间段的细胞存活率并计算出细胞抑制率与半数抑制浓度(IC50),绘制细胞抑制百分率曲线;用试剂盒检测各组细胞丙二醛(MDA)含量、超氧化物歧化酶(SOD)及乳酸脱氢酶(LDH)活性;采用流式细胞术检测各组细胞活性氧(ROS)含量和细胞凋亡率。[结果]纳米二氧化硅染毒A549细胞24、48 h后,随着染毒浓度的升高,细胞形态发生改变,细胞出现皱缩、空泡。染毒24、48 h后,与对照组比较,各剂量组A549细胞抑制率均明显增加(P0.05),IC50分别为125.8、50.83 mg/L。染毒24、48 h后,与对照组相比,各浓度染毒组MDA含量、LDH活性随着染毒浓度的升高而升高(P0.05),而SOD活性随着染毒浓度升高呈下降趋势(P0.05)。ROS含量与凋亡率检测结果显示,与对照组相比,染毒24、48 h的染毒组中ROS与细胞凋亡率随着染毒浓度升高而升高,差异具有统计学意义(P0.05)。[结论]纳米二氧化硅能够损伤A549细胞的细胞膜,产生氧化应激反应并刺激ROS的释放,引起细胞凋亡甚至死亡。
[Abstract]:[objective] to study the effects of nano-silica on oxidative stress and apoptosis of human lung adenocarcinoma A549 cells. [methods] A549 cells were exposed to nano-silica solution of 25,50100200 mg/L for 24 and 48 hours, but the control group was not treated. The morphological changes of cells were observed by fluorescence microscope. Cell count kit (cell counting kit,CCK-8) was used to detect the survival rate of each group and the cell inhibition rate and 50% inhibitory concentration (IC50) were calculated, and the percentage curve of cell inhibition was drawn. Malondialdehyde (MDA) content, superoxide dismutase (SOD) (SOD) and lactate dehydrogenase (LDH) activity were measured with kit, and reactive oxygen species (Ros) (ROS) content and apoptosis rate were measured by flow cytometry (FCM). [results] after exposure to nano-silica for 24 h and 48 h, the morphology of A549 cells was changed with the increase of the concentration of nano-Sio _ 2, and the cells were shriveled and vacuolated. After 24 h and 48 h treatment, the inhibition rates of A549 cells were significantly increased compared with the control group (P0.05), the IC50 were 125.8 mg/L. and 50.83 mg/L., respectively. After 24 h, 48 h, compared with the control group, the content of MDA and the activity of LDH in each concentration group increased with the increase of the concentration (P0.05). The activity of SOD decreased with the increase of exposure concentration (P 0.05). The results of Ros content and apoptosis rate showed that compared with the control group, ROS and apoptosis rate increased with the increase of exposure concentration at 48 h. The difference was statistically significant (P 0.05). [conclusion] Nano-silica can damage the cell membrane of A549 cells, produce oxidative stress reaction, stimulate the release of ROS, and induce apoptosis and even death of A549 cells.
【作者单位】：东南大学公共卫生学院苏州纳米科技协同创新中心环境医学工程教育部重点实验室江苏省生物材料与器件重点实验室;
【基金】：国家重点基础研究发展计划项目(编号:2011CB933404) 国家自然科学基金项目(编号:81302461,81473003,81502783,81573186,31671034,81673218) 江苏省研究生创新人才培养基金(编号:SJLX15_0072)
【分类号】：R114

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