Zhang Li, Zhang Yong-Qiang, Li Xi, Wu Shuai, Yang Dan-Feng
Institute of Environmental and Operational Medicine, Institute of Military Medilal Sciences, Academy of Military Sciences, Tianjin 300050, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Sep;36(5):394-399. doi: 10.12047/j.cjap.5983.2020.084.
To investigate the effects of nano-SiO and cold on the cytotoxicity and secretion of inflammatory factors in human lung adenocarcinoma epithelial cell line A549. A549 was used as experimental subject, a single factor multilevel experiment was designed, A549 cells were exposed to 10, 50, 100, 200 μg/ml nano-SiO particles and/or cultured at 31℃, 33℃, 35℃ for 48 h. After that, cell morphology was observed and relative cell survival rate was detected. According to the results of single factor analysis and based on the selection of nano-SiO dose and temperature that significantly reduced the relative survival rate of A549 cells, the experiment was designed according to 2×2 factor analysis , they were divided into 4 groups: control group(37℃), Nano-SiO exposure group, low temperature exposure group, Nano-SiO and low temperature composite group. After exposure for 48 h, the supernatant of cell culture medium was collected for detecting the LDH activity by colorimetric method and the levels of cytokines interleukin-6 (IL-6) and interleukin-8 (IL-8) were determined by ELISA. The mRNA levels of cellular IL-6 and IL-8 were detected by qRT-PCR. The activities of A549 cells in 100 μg/ml Nano-SiO group and 31℃ low temperature group were decreased significantly. Under the combined conditions, the activity of A549 cells was most inhibited (<0.01), and the levels of inflammatory factors IL-6 and IL-8 and mRNA were significantly increased (<0.01). 100 μg/ml Nano-SiO combined with 31℃ cold exposure can synergistically reduce the activity of A549 cells and increase the expression level of inflammatory factors IL-6 and IL-8 .
为研究纳米二氧化硅(nano-SiO)与寒冷对人肺腺癌上皮细胞系A549细胞毒性及炎性因子分泌的影响。以A549细胞为实验对象,设计单因素多水平实验,将A549细胞分别暴露于10、50、100、200μg/ml纳米二氧化硅颗粒及/或在31℃、33℃、35℃培养48 h。之后,观察细胞形态并检测相对细胞存活率。根据单因素分析结果,选取能显著降低A549细胞相对存活率的纳米二氧化硅剂量和温度,按2×2析因分析设计实验,分为4组:对照组(37℃)、纳米二氧化硅暴露组、低温暴露组、纳米二氧化硅与低温复合组。暴露48 h后,收集细胞培养基上清液,采用比色法检测乳酸脱氢酶(LDH)活性,采用酶联免疫吸附测定法(ELISA)测定细胞因子白细胞介素-6(IL-6)和白细胞介素-8(IL-8)水平。采用实时荧光定量聚合酶链反应(qRT-PCR)检测细胞IL-6和IL-8的mRNA水平。100μg/ml纳米二氧化硅组和31℃低温组A549细胞活性显著降低。在联合条件下,A549细胞活性受到的抑制最为明显(P<0.01),炎性因子IL-6和IL-8及其mRNA水平显著升高(P<0.01)。100μg/ml纳米二氧化硅与31℃寒冷暴露联合可协同降低A549细胞活性,增加炎性因子IL-6和IL-8的表达水平。