梁晓瑜, 张小强, 张妍, 刘静, 王旭. 纳米氧化锌对小胶质瘤BV2细胞炎症因子及p38MAPK蛋白磷酸化的影响[J]. 环境与职业医学, 2018, 35(5): 447-451. DOI: 10.13213/j.cnki.jeom.2018.17684
引用本文: 梁晓瑜, 张小强, 张妍, 刘静, 王旭. 纳米氧化锌对小胶质瘤BV2细胞炎症因子及p38MAPK蛋白磷酸化的影响[J]. 环境与职业医学, 2018, 35(5): 447-451. DOI: 10.13213/j.cnki.jeom.2018.17684
LIANG Xiao-yu, ZHANG Xiao-qiang, ZHANG Yan, LIU Jing, WANG Xu. Effects of zinc oxide nanoparticle on expression of inflammatory factors and phosphorylation of p38MAPK in BV2 microglia[J]. Journal of Environmental and Occupational Medicine, 2018, 35(5): 447-451. DOI: 10.13213/j.cnki.jeom.2018.17684
Citation: LIANG Xiao-yu, ZHANG Xiao-qiang, ZHANG Yan, LIU Jing, WANG Xu. Effects of zinc oxide nanoparticle on expression of inflammatory factors and phosphorylation of p38MAPK in BV2 microglia[J]. Journal of Environmental and Occupational Medicine, 2018, 35(5): 447-451. DOI: 10.13213/j.cnki.jeom.2018.17684

纳米氧化锌对小胶质瘤BV2细胞炎症因子及p38MAPK蛋白磷酸化的影响

Effects of zinc oxide nanoparticle on expression of inflammatory factors and phosphorylation of p38MAPK in BV2 microglia

  • 摘要: 目的 探讨纳米氧化锌(zinc oxide nanoparticle,nano-ZnO)对小胶质瘤BV2细胞炎症因子及p38丝裂原活化蛋白激酶(p38MAPK)磷酸化的影响。

    方法 不同质量浓度nano-ZnO(0、5.0、10.0、15.0、20.0、30.0、50.0、75.0 mg/L)染毒小胶质瘤BV2细胞24 h,用MTT法测定细胞存活率后确定染毒剂量。以nano-ZnO(0、5.0、10.0、15.0、20.0 mg/L)染毒BV2细胞24 h,采用乳酸脱氢酶(LDH)检测试剂盒检测细胞培养液上清液中LDH活性,ELISA检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的分泌水平,Western blot法检测磷酸化p38MAPK(p-p38MAPK)蛋白水平并用Image J软件对蛋白条带灰度值进行分析。应用Spearman相关分析炎症因子分泌与p38MAPK蛋白磷酸化水平的相关性。

    结果 随nano-ZnO染毒剂量的增加,BV2细胞存活率降低(r=-0.994,P < 0.001),细胞培养液上清液中LDH活性增加(r=0.749,P < 0.001)。与对照组相比,10.0、15.0、20.0mg/L nano-ZnO染毒组TNF-α、IL-1β、IL-6分泌水平增加(均Ps < 0.05)。Western blot检测结果发现各染毒组p-p38MAPK蛋白表达水平增高,且与炎症因子分泌水平的变化趋势一致(rTNF-α=0.836,P < 0.001;rIL-6=0.539,P < 0.001;rIL-1β=0.659,P=0.008);20.0 mg/L nano-ZnO染毒组p-p38MAPK与p38MAPK灰度值的比值较对照组高,差异有统计学意义(P < 0.05)。

    结论 nano-ZnO可诱导BV2细胞炎症因子分泌水平和p38MAPK蛋白磷酸化水平增高。

     

    Abstract: Objective To investigate the effects of zinc oxide nanoparticle (nano-ZnO) on expression of inflammatory factors and phosphorylation of p38 mitogen-activated protein kinases (p38MAPK) in BV2 microglia.

    Methods BV2 cells were treated with different concentrations of nano-ZnO (0, 5.0, 10.0, 15.0, 20.0, 30.0, 50.0, and 75.0 mg/L) for 24 h. MTT assay was used to assess the viability of BV2 cells and determine the exposure concentration. Then, BV2 microglia were treated with nano-ZnO (0, 5.0, 10.0, 15.0, and 20.0 mg/L) for 24 h, and the levels of lactate dehydrogenase (LDH) was detected by LDH detection kit, the secretion of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, and IL-6 by ELISA, the protein levels of phosphorylated p-38MAPK (p-p38MAPK) by Western Blot, and the gray value of protein band by Image J software. The relationship between the expression of inflammatory factors and the protein level of p-p38MAPK was analyzed by Spearman correlation analysis.

    Results Increasing nano-ZnO treatment significantly decreased the viability of BV2 cells (r=-0.994, P < 0.001) and increased the activity of LDH (r=0.749, P < 0.001). Compared to the control group, the levels of TNF-α, IL-1β, and IL-6 increased in the 10.0, 15.0, and 20.0 mg/L nano-ZnO exposure groups (Ps < 0.05). Western bolt results showed that the expression levels of p-p38MAPK increased in the nano-ZnO exposure groups and were consistent with the change trend of inflammatory factor levels (rTNF-α=0.836, P < 0.001; rIL-6=0.539, P < 0.001; rIL-1β=0.659, P=0.008). The grey value ratio of p-p38MAPK and p38MAPK was higher in the 20.0mg/L nano-ZnO exposure group than in the control group (P < 0.05).

    Conclusion Exposure to nano-ZnO could induce elevated secretion levels of inflammatory cytokines and expression levels of p38MAPK phosphorylation in BV2 microglia.

     

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