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*

doi:10.12047/j.cjap.5983.2020.084

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

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^△

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^△

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/Abstract

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-SiO₂dose 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-SiO₂group 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

中图分类号:

R122

张莉, 张永强, 李曦, 武帅, 杨丹凤. 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^*[J]. 中国应用生理学杂志, 2020, 36(5): 394-399.

ZHANG Li, ZHANG Yong-qiang, LI Xi, WU Shuai, YANG Dan-feng. Effects of Nano-SiO₂ combining with cold on cytotoxicity and secretion of inflammatory factors in A549 cells[J]. CJAP, 2020, 36(5): 394-399.

参考文献

[1] Liu L, Urch B, Poon R, et al. Effects of ambient coarse, fine, and ultrafine particles and their biological constituents on systemic biomarkers: a controlled human exposure study[J]. Environ Health Persp, 2015, 123(6): 534-540.
[2] Günter O, Eva O, Jan Or. Nanotoxicology: An emerging discipline evolving from studies of ultrafine particles[J]. Environ Health Persp, 2005, 118(9): A380.
[3] Dai L, Zanobetti A, Koutrakis P, et al. Associations of fine particulate matter species with mortality in the United States: A multicity time-series analysis[J]. Environ Health Persp, 2014, 122(8): 837.
[4] Wen D. Nanofuel as a potential secondary energy carrier[J]. Energ Environ Sci, 2010, 3(5): 591-600.
[5] Kaewamatawong T, Shimada A, Okajima M, et al. Acute and subacute pulmonary toxicity of low dose of ultrafine colloidal silica particles in mice after intratracheal instillation[J]. Toxicol Pathol, 2006, 34(7): 958-965.
[6] Nemmar A, Vanbilloen H, Hoylaerts MF, et al. Passage of intratracheally instilled ultrafine particles from the lung into the systemic circulation in hamster[J]. Am J Respir Crit Care Med, 2001, 164(9): 1665-1668.
[7] Claudia FP. The toxicological mode of action and the safety of synthetic amorphous silica—A nanostructured material[J]. Toxicology, 2012, 294(2-3): 61-79.
[8] Jaakkola K, Saukkoriipi A, Jokelainen J, et al. Decline in temperature and humidity increases the occurrence of influenza in cold climate[J]. Environ Health, 2014, 13(1): 22.
[9] Carder M, McNamee R, Beverland I, et al. Interacting effects of particulate pollution and cold temperature on cardiorespiratory mortality in Scotland[J]. Occup Environ Med, 2008, 65(3): 197-204.
[10]Li Y, Ma Z, Zheng C, et al. Ambient temperature enhanced acute cardiovascular-respiratory mortality effects of PM2.5 in Beijing, China[J]. Int J Biometeorol, 2015, 59(12): 1761-1770.
[11]Qian Z, He Q, Lin H-M, et al. High temperatures enhanced acute mortality effects of ambient particle pollution in the "Oven" city of Wuhan, China[J]. Environ Health Persp, 2008, 116(9): 1172-1178.
[12]徐雯, 江俊康, 杨北平, 等. 不同粒径汽车尾气颗粒物对A549细胞毒性作用的比较[J]. 环境科学学报, 2013, 33(12): 3407-3412.
[13]Chang JS, Chang KL, Hwang DF, et al. In vitro cytotoxicitiy of silica nanoparticles at high concentrations strongly depends on the metabolic activity type of the cellline[J]. Environ Sci Technol, 2007, 41(6): 2064-2068.
[14]Sabnis AS, Shadid M, Yost GS, et al. Human lung epithelial cells express a functional cold-sensing TRPM8 variant[J]. Am J Resp Cell Mol, 2008, 39(4): 466.
[15]Halatek T, Stepnik M, Stetkiewicz J, et al. The inflammatory response in lungs of rats exposed on the airborne particles collected during different seasons in four European cities[J]. Environ Let, 2011, 46(13): 1469-1481.
[16]VanEeden SF, Tan WC, Suwa T, et al. Cytokines involved in the systemic inflammatory response induced by exposure to particulate matter air pollutants(PM10)[J]. Am J Resp Crit Care, 2001, 164(5): 826.
[17]卢志刚, 王亚利, 张明泉, 等. 秋冬季节对健康大鼠肺组织表面活性蛋白A和白细胞介素6表达的影响[J]. 中国组织工程研究, 2010, 14(11): 2000-2003.
[18]Zhang Y, Lin Y, Xi L, et al. Silica dioxide nanoparticles combined with cold exposure induce stronger systemic inflammatory response[J]. Environ Sci Pollut R, 2016, 24(1): 291-298.
[19]张士涵, 梁雪, 汪岩, 等. 纳米二氧化硅对体外培养A549细胞氧化应激与凋亡的影响[J]. 环境与职业医学, 2017, 34(5): 449-453.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 6

编辑推荐

Metrics

本文评价

[1]	方振, 胡西厚, 李康, 韩洁, 田蕾, 闫峻, 张伟, 来文庆, 林本成, 刘晓华, 袭著革. PM_2.5鼻腔滴注致小鼠海马组织损伤的炎性机制[J]. 中国应用生理学杂志, 2020, 36(3): 240-244.
[2]	赵海, 李曦, 张莉, 武帅, 张永强, 李军, 张格祥, 杨丹凤. 寒冷对雌性C57BL/6小鼠动情周期的影响^*[J]. 中国应用生理学杂志, 2019, 35(5): 454-456.
[3]	张瑞奎, 汪超. 苦参碱对大鼠乳腺癌细胞荷瘤生长及其炎性因子与免疫功能的影响[J]. 中国应用生理学杂志, 2018, 34(4): 375-378.
[4]	赵丹丹, 郭展, 李曦, 张永强, 张莉, 朱五福, 郑鹏武, 杨丹凤. 纳米炭黑颗粒复合寒冷暴露对小鼠肺部氧化应激反应的影响[J]. 中国应用生理学杂志, 2018, 34(3): 258-262.
[5]	罗梓垠, 高慧, 项冰倩, 邱晓晓, 戴雍月, 王万铁. 右美托咪定对A549细胞缺氧/复氧损伤时细胞凋亡及CHOP的影响[J]. 中国应用生理学杂志, 2018, 34(2): 137-142.
[6]	张慧苹, 马丽千, 吕艺, 罗红敏, 李雨梦, 刘锐, 胡森. 二甲亚砜对酵母多糖致肠道屏障损伤的保护作用[J]. 中国应用生理学杂志, 2017, 33(3): 193-196.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 6

编辑推荐

Metrics

本文评价

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