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亚砷酸钠对人胚肺成纤维细胞核因子κB信号通路的影响及其作用机制

Effects of sodium arsenite on nuclear factor-κB signaling pathway in human embryonic lung fibroblasts

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【作者】 张曦小雪王素华赵宇航高艳荣

【Author】 ZHANGXI Xiao-xue;WANG Su-hua;ZHAO Yu-hang;GAO Yan-rong;School of Public Health,Baotou Medical College of Inner Mongolia University of Science and Technology;

【通讯作者】 王素华;

【机构】 内蒙古科技大学包头医学院公共卫生学院

【摘要】 目的探讨核因子κB(NF-κB)信号通路在亚砷酸钠染毒致人胚肺成纤维细胞(HELF)损伤中的作用机制。方法将HELF细胞分别暴露于0(对照)、0.625、1.25、2.5、5、10、20、40μmol/L亚砷酸钠溶液染毒18、24、48 h,采用MTT法检测细胞存活率。将HELF细胞分别暴露于0(对照)、0.625、1.25、2.5、5、10、20、40μmol/L亚砷酸钠溶液染毒24 h,采用ELISA法检测上清液中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和白介素-1β(IL-1β)炎性因子的浓度。将HELF细胞分为2组,一组分别暴露于0(对照)、2.5、5、10、20μmol/L的亚砷酸钠溶液,另一组同时加入10μmol/L的NF-κB抑制剂PDTC溶液,染毒24 h后,采用Western blot法检测细胞NF-κB相关蛋白p-IKKβ和p-P65的表达水平。结果与对照组比较,2.5、5、10、20、40μmol/L亚砷酸钠染毒HELF细胞18、24、48 h后的细胞存活率均下降,除2.5 mol/L亚砷酸钠染毒48 h外,差异均有统计学意义(P<0.05);且随着亚砷酸钠染毒浓度的升高和染毒时间的延长,各染毒时间HELF细胞的存活率均呈下降趋势(均P<0.01)。与对照组比较,1.25、2.5、5、10、20、40μmol/L亚砷酸钠染毒组HELF细胞上清液中TNF-α的浓度均升高,5、10、20、40μmol/L亚砷酸钠染毒组HELF细胞上清液中IL-6的浓度均升高,各浓度亚砷酸钠染毒组HELF细胞上清液中IL-1β的浓度均升高,差异均有统计学意义(P<0.05);且随着亚砷酸钠染毒浓度的升高,HELF细胞上清液中TNF-α、IL-6、IL-1β的浓度均呈上升趋势。与对照组和相同浓度PDTC干预组比较,各浓度亚砷酸钠染毒组HELF细胞p-IKKβ和p-P65蛋白的表达水平均升高,差异有统计学意义(P<0.05)。结论亚砷酸钠暴露可激活NF-κB通路,促使炎性因子增加,从而造成HELF细胞损伤。

【Abstract】 Objective To investigate the mechanism of nuclear factor κB(NF-κB) signaling pathway in the damage of human embryonic lung fibroblasts(HELF) induced by sodium arsenite. Methods HELF cells were exposed to sodium arsenite at the doses of 0(control group),0.625,1.25,2.5,5,10,20 and 40 μmol/L respectively for 24 h,and ELISA was used to detect concentration of inflammatory factors TNF-α,IL-6 and IL-1β in the supernatant. The HELF cells were divided into two groups,one group was exposed to sodium arsenite at the doses of 0(control group),2.5,5,10 and 20 μmol/L,and the other group was added 10 μmol/L pyrrolidine dithiocarbamate(PDTC),the inhibitor of NF-κB,and exposed to the same doses of sodium arsenite. After 24 h of exposure,the expression levels of NF-κB-related proteins p-IKKβ and p-P65 were detected by Western blot. Results Compared with the control group,the concentration of TNF-α in the supernatant of HELF cells exposed to 1.25-40 μmol/L sodium arsenite increased,the concentration of IL-6 in 5-40 μmol/L sodium arsenite groups increased,the concentration of IL-1β in all sodium arsenite exposure groups increased(P<0.05) with a dose dependent trend. Compared with the control group and the PDTC intervention group exposed to the same concentration of sodium arsenite,the expression level of p-IKKβ and p-P65 protein in HELF cells in all sodium arsenite exposure groups increased significantly(P<0.05). Conclusion Sodium arsenite may activate the NF-κB signaling pathway and promote inflammatory factors production,resulting in HELF cell damage.

【基金】 国家自然科学基金(81660532)
  • 【文献出处】 环境与健康杂志 ,Journal of Environment and Health , 编辑部邮箱 ,2018年10期
  • 【分类号】R114
  • 【下载频次】42
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