王会敏, 陈涛, 童建, 姜岩. PM2.5染毒干扰心肌细胞分化发育的体外实验[J]. 环境与职业医学, 2018, 35(6): 536-540. DOI: 10.13213/j.cnki.jeom.2018.17744
引用本文: 王会敏, 陈涛, 童建, 姜岩. PM2.5染毒干扰心肌细胞分化发育的体外实验[J]. 环境与职业医学, 2018, 35(6): 536-540. DOI: 10.13213/j.cnki.jeom.2018.17744
WANG Hui-min, CHEN Tao, TONG Jian, JIANG Yan. Disruption of cardiac differentiation by PM2.5 in vitro[J]. Journal of Environmental and Occupational Medicine, 2018, 35(6): 536-540. DOI: 10.13213/j.cnki.jeom.2018.17744
Citation: WANG Hui-min, CHEN Tao, TONG Jian, JIANG Yan. Disruption of cardiac differentiation by PM2.5 in vitro[J]. Journal of Environmental and Occupational Medicine, 2018, 35(6): 536-540. DOI: 10.13213/j.cnki.jeom.2018.17744

PM2.5染毒干扰心肌细胞分化发育的体外实验

Disruption of cardiac differentiation by PM2.5 in vitro

  • 摘要: 目的 研究PM2.5染毒对心肌细胞分化发育的影响。

    方法 利用P19畸胎瘤干细胞,建立体外心肌定向分化模型,MTT法检测PM2.5染毒对P19细胞毒性,PM2.5染毒浓度为0、1、10、100 mg/L,选择未产生明显细胞毒性的10 mg/L PM2.5于P19细胞未分化时期染毒48 h后启动分化。显微镜下观察形态学改变,实时荧光定量PCR检测不同分化期OCT4ISL-1MLC2V的表达变化(OCT4ISL-1MLC2V分别为干细胞全能性阶段、心肌前体期、成熟心肌期标志基因),免疫荧光及流式细胞术检测成熟心肌肌钙蛋白cTNT的表达效率。

    结果 与对照组相比,PM2.5组细胞形态学发生改变;分化各阶段的标志物表达异常:OCT4在未分化时期表达降低约80%(P < 0.01),分化启动后反而增高(P < 0.05);ISL-1于心肌前体期表达降低为对照组的59%(P < 0.01);MLC2V于成熟细胞期表达降低为对照组的39%(P < 0.01)。成熟心肌cTNT表达比例由对照组的61%下降到47%,但差异无统计学意义。

    结论 心肌分化早期染毒PM2.5,可阻碍干细胞向心肌细胞的分化过程。

     

    Abstract: Objective To study the effect of PM2.5 exposure on cardiac differentiation.

    Methods P19 mouse carcinoma stem cells were used as an in vitro cardiac differentiation model, and MTT assay was performed to examine the cytoplasmic toxicity of PM2.5 exposure at different concentrations (0, 1, 10, and 100 mg/L). The cells were exposed to PM2.5 at the non-cytotoxic concentration of 10 mg/L for 48 h before the differention was initiated. After the differentiation was initiated, the morphological changes were observed, the expression changes of cardiac specific genes (OCT4, pluripotent gene; ISL-1, precursor formation related gene; MLC2V, highly expressed at maturation period) were evaluated by real-time fluorescence quantitative PCR, and the expression efficiency of cardiac functional protein cardiac troponin T (cTNT) by immunofluorescence and flow cytometry.

    Results Compared with the control group, the PM2.5 groups showed cell morphological changes and the cardiac specific genes were abnormally expressed across all differentiation stages. OCT4 expression was reduced by 80% before differentiation (P < 0.01), and increased after differentiation (P < 0.05); ISL-1 expression was reduced to 59% of the control group in precursor formation period (P < 0.01); MLC2V expression was reduced to 39% of the control group in maturation period (P < 0.01). Besides, the cardiac functional protein cTNT decreased its expression from 61% to 47% after PM2.5 exposure, but there was no difference.

    Conclusion Early exposure to PM2.5 during the cardiac development may disturb cardiac differentiation of stem cells.

     

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