郭旺旺, 刘进仁, 范小琳, 王鸿, 牛浩, 高俊宏. 奥克托今及其主要合成中间体对中国仓鼠肺细胞凋亡和氧化应激的影响[J]. 环境与职业医学, 2020, 37(8): 808-811. DOI: 10.13213/j.cnki.jeom.2020.20083
引用本文: 郭旺旺, 刘进仁, 范小琳, 王鸿, 牛浩, 高俊宏. 奥克托今及其主要合成中间体对中国仓鼠肺细胞凋亡和氧化应激的影响[J]. 环境与职业医学, 2020, 37(8): 808-811. DOI: 10.13213/j.cnki.jeom.2020.20083
GUO Wang-wang, LIU Jin-ren, FAN Xiao-lin, WANG Hong, NIU Hao, GAO Junhong. Effects of octogen and its intermediate product on apoptosis and oxidative stress of Chinese hamster lung cells[J]. Journal of Environmental and Occupational Medicine, 2020, 37(8): 808-811. DOI: 10.13213/j.cnki.jeom.2020.20083
Citation: GUO Wang-wang, LIU Jin-ren, FAN Xiao-lin, WANG Hong, NIU Hao, GAO Junhong. Effects of octogen and its intermediate product on apoptosis and oxidative stress of Chinese hamster lung cells[J]. Journal of Environmental and Occupational Medicine, 2020, 37(8): 808-811. DOI: 10.13213/j.cnki.jeom.2020.20083

奥克托今及其主要合成中间体对中国仓鼠肺细胞凋亡和氧化应激的影响

Effects of octogen and its intermediate product on apoptosis and oxidative stress of Chinese hamster lung cells

  • 摘要: 背景

    奥克托今(HMX)是当今综合性能最好的单质炸药。有研究发现HMX可影响职业暴露者神经行为功能和周围神经传导速度。3,7-二硝基-1,3,5,7-四氮杂双环3,3,1壬烷(DPT)是合成HMX的主要中间体,但目前国内外关于DPT的毒性研究尚未见报道。

    目的

    探讨HMX及DPT对中国仓鼠肺细胞(CHL)细胞凋亡及氧化应激水平的影响。

    方法

    应用含有0、31.25、62.50、125、250、500 mg·L-1质量浓度(后称浓度)的HMX、DPT分别对体外培养的CHL染毒24 h,利用CCK-8法检测细胞活力;Annexin V-FITC/PI双染法检测细胞凋亡情况;DCFH-DA荧光探针检测细胞内活性氧(ROS)荧光强度。采用抗氧化剂干预,即2.5 mmol·L-1 N-乙酰半胱氨酸(NAC)预处理CHL 4 h,经120 mg·L-1 DPT染毒24 h后检测细胞活力和凋亡情况。

    结果

    细胞活力检测结果显示DPT对CHL的半数抑制浓度为119.55 mg·L-1。当DPT浓度为250 mg·L-1时,细胞活力下降至(17.95±2.18)%;HMX浓度为500 mg·L-1时,细胞活力为(83.34±3.03)%。细胞凋亡结果显示,与各自对照组相比,各染毒组的凋亡率均随染毒剂量增加而逐渐升高(P < 0.05)。62.50~500 mg·L-1范围内,HMX导致的细胞凋亡率由(15.63±0.58)%缓慢升至(37.00±1.25)%,而DPT引起的细胞凋亡率从(33.75±0.57)%迅速升至(96.57±0.53)%。细胞内ROS检测结果显示,HMX和DPT均能引起细胞内ROS水平升高。但HMX引起细胞内ROS变化较小,250、500 mg·L-1 HMX组细胞内ROS荧光强度分别为103.96±5.59和119.61±4.42,而相同剂量DPT组的ROS荧光强度分别为238.52±7.15和451.02±13.02(P < 0.05)。另外,NAC干预可有效缓解DPT引起的细胞活力抑制和凋亡,NAC预处理+DPT组的细胞活力、凋亡率为(76.41±4.91)%、(16.85±0.12)%,而DPT组为(48.02±2.63)%、(62.67±8.49)%(P < 0.05)。

    结论

    相同剂量下DPT对CHL的毒性大于HMX,高剂量DPT引起细胞内ROS水平升高,导致细胞凋亡。

     

    Abstract: Background

    Octogen (HMX) is the best comprehensive explosive compound at present. Studies have found that HMX can affect the neurobehavioral function and peripheral nerve conduction velocity of occupational exposure groups. 3, 7-dinitro-1, 3, 5, 7-tetraazabicyclo3,3,1nonane (DPT) is the main intermediate of HMX synthesis, but the toxicities of DPT have been rarely reported across the world.

    Objective

    This experiment investigates the effects of HMX and DPT on apoptosis and oxidative stress of Chinese hamster lung cells (CHL).

    Methods

    Degsined doses (0, 31.25, 62.50, 125, 250, and 500 mg·L-1) of HMX and DPT were applied to treat CHL cultured in vitro for 24 h. CCK-8 method was used to detect cell viability, Annexin V-FITC/PI double staining assay for cell apoptosis, and DCFH-DA fluorescent probe for intracellular reactive oxygen species (ROS). Furthermore, having been pretreated with 2.5 mmol·L-1 N-acetyl cysteine (NAC) for 4h and administered with 120 mg·L-1 DPT for 24h, the cell viability and apoptosis of CHL were detected.

    Results

    The results of cell viability assay showed that the half inhibitory concentration of DPT on CHL was 119.55 mg·L-1. DPT at 250 mg·L-1 decreased the cell viability to (17.95±2.18)%, and 500 mg·L-1 HMX decreased it to (83.34±3.03)%. Compared with corresponding control groups, the apoptosis rates of the exposure groups increased gradually with the exposure dose rising (P < 0.05). The apoptosis rate of the 62.50-500 mg·L-1 HMX exposure groups increased moderately from (15.63±0.58)% to (37.00±1.25)%, while that of the 62.50-500 mg·L-1 DPT exposure groups increased drastically from (33.75±0.57)% to (96.57±0.53)%. Both HMX and DPT increased intracellular ROS levels. However, HMX caused a smaller change of ROS levels than DPT did. The ROS intensity were 103.96±5.59 and 119.61±4.42 at 250 and 500 mg·L-1 HMX, and 238.52±7.15 and 451.02±13.02 at 250 and 500 mg·L-1 DPT, respectively (P < 0.05). In addition, the NAC intervention effectively alleviated the cell viability inhibition and apoptosis caused by the DPT exposure. The cell viability and apoptosis rate of the NAC pretreatment+DPT group were (76.41±4.91)% and (16.85±0.12)%, while those of the DPT group were (48.02±2.63)% and (62.67±8.49)%, respectively (P < 0.05).

    Conclusion

    The toxicity of DPT to CHL is higher than that of HMX at the same dose. DPT at a high concentration could increase cell apoptosis of CHL via elevating intracellular ROS.

     

/

返回文章
返回