纳米二氧化硅复合寒冷对A549细胞毒性及其炎性因子分泌的影响

Effects of Nano-SiO_(2) combining with cold on cytotoxicity and secretion of inflammatory factors in A549 cells

目的:本研究旨在探讨纳米二氧化硅(Nano-SiO_(2))颗粒和寒冷复合对人肺腺癌上皮细胞A549细胞毒性及炎性因子分泌的影响。方法:本研究以A549细胞为实验对象,分别用10,50,100,200μg/ml Nano-SiO_(2)颗粒对A549细胞染毒,以及分别在35℃,33℃,31℃条件下对A549细胞进行低温暴露,培养48 h后,观察细胞形态及测定细胞相对存活率。根据单因素分析结果,选出对A549细胞相对存活率有显著降低作用的Nano-SiO_(2)剂量和温度的基础上,按照2×2析因设计实验,分为4组:①37℃对照组;②Nano-SiO_(2)染毒组;③低温暴露组;④Nano-SiO_(2)和低温复合组,不同条件下暴露48 h后,收集细胞上清液采用比色法检测LDH活性,以及ELISA法测定细胞因子白介素-6(IL-6)和白介素-8(IL-8)的水平,采用qRT-PCR法检测细胞IL-6和IL-8的基因表达水平。结果:100μg/ml Nano-SiO_(2)组和31℃低温组能够显著降低A549细胞活性(P<0.01),在复合条件作用下对A549细胞活性抑制最为显著,且炎性因子IL-6和IL-8及mRNA的表达水平均显著升高(P<0.01)。结论:100μg/ml Nano-SiO_(2)与31℃低温复合暴露可协同降低A549细胞的相对存活率,增加炎性因子IL-6和IL-8表达水平。

Objective:To investigate the effects of nano-SiO_(2) and cold on the cytotoxicity and secretion of inflammatory factors in human lung adenocarcinoma epithelial cell line A549.Methods:A549 was used as experimental subject,a single factor multilevel experiment was designed,A549 cells were exposed to 10,50,100,200μg/ml nano-SiO_(2) 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_(2) 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_(2) exposure group,low temperature exposure group,Nano-SiO_(2) 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.Results:The activities of A549 cells in 100μg/ml Nano-SiO_(2) group and 31℃low temperature group were decreased significantly.Under the combined conditions,the activity of A549 cells was most inhibited(P<0.01),and the levels of inflammatory factors IL-6 and IL-8 and mRNA were significantly increased(P<0.01).Conclusion:100μg/ml Nano-SiO_(2) 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.