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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (5): 394-399.doi: 10.12047/j.cjap.5983.2020.084

• 研究论文 • 上一篇    下一篇

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*

张莉, 张永强, 李曦, 武帅, 杨丹凤   

  1. 军事科学院军事医学研究院环境医学与作业医学研究所, 天津 300050
  • 收稿日期:2019-12-02 修回日期:2020-07-17 发布日期:2021-02-25
  • 通讯作者: Tel: 022-84655058; E-mail: fengdyd@126.com
  • 基金资助:
    *国家自然科学基金青年项目资助(21707165);BWS16J010

Effects of Nano-SiO2 combining with cold on cytotoxicity and secretion of inflammatory factors in A549 cells

ZHANG Li, ZHANG Yong-qiang, LI Xi, WU Shuai, YANG Dan-feng   

  1. Institute of Environmental and Operational Medicine, Institute of Military Medilal Sciences, Academy of Military Sciences, Tianjin 300050, China
  • Received:2019-12-02 Revised:2020-07-17 Published:2021-02-25

摘要: 目的: 本研究旨在探讨纳米二氧化硅(Nano-SiO2)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, 200 μg/ml Nano-SiO2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-SiO2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-SiO2染毒组;③低温暴露组;④Nano-SiO2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: 100 μg/ml Nano-SiO2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: 100 μg/ml Nano-SiO2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。

关键词: 寒冷, 纳米二氧化硅, A549细胞, 细胞毒性, 炎性因子

Abstract: Objective: To investigate the effects of nano-SiO2 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, 200 μg/ml nano-SiO2 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-SiO2 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-SiO2 exposure group, low temperature exposure group, Nano-SiO2 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 100 μg/ml Nano-SiO2 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: 100 μg/ml Nano-SiO2 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 .

Key words: cold, Nano-SiO2, A549 cells, cellular activity, inflammatory factors

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