Fine particulate matter reduces the pluripotency and proliferation of human embryonic stem cells through ROS induced AKT and ERK signaling pathway
Fine particulate matter reduces the pluripotency and proliferation of human embryonic stem cells through ROS induced AKT and ERK signaling pathway
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细颗粒物通过ROS诱导的AKT和ERK信号通路降低人胚胎干细胞的多能性和增殖
DOI：
10.1016/j.reprotox.2020.07.010
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发表时间：
2020-09-01
影响因子：
3.3
通讯作者：
Du, Lili
中科院分区：
文献类型：
--
作者：
Bi, Shilei;Tang, Jingman;Du, Lili
流行病学调查发现，暴露在空气中的细颗粒物(PM)不仅会导致成人和儿童的呼吸系统和心血管疾病，还会影响怀孕期间的胚胎发育，导致不良的妊娠结局。然而，其确切的分子机制仍不清楚。本研究用不同浓度的PM处理人胚胎干细胞(HESCs)，观察其形态和增殖能力。用定量聚合酶链式反应、免疫荧光、免疫印迹和流式细胞仪检测NANOG和OCT4的mRNA和蛋白表达。同时检测细胞内活性氧(ROS)生成和AKT/ERK活性。同时，在PM暴露前加入或不加入ROS清除剂N-乙酰半胱氨酸(NAC)，检测HESCs中ROS、NANOG、OCT4的表达以及AKT/ERK信号通路的变化。PM暴露后，细胞增殖能力及OCT4和NANOG在mRNA和蛋白水平的表达均下调。在PM暴露后，HESCs中的ROS水平增加，但这种增加可被NAC预处理所减弱。进一步分析表明，PM暴露后，AKT和ERK的磷酸化水平升高。经NAC处理后，对hESC的增殖、多能性和分化起重要调控作用的AKT和ERK的磷酸化水平明显低于未经NAC处理的暴露组。这些结果表明，PM暴露可能通过ROS介导的AKT/ERK途径降低hESC的增殖和多能性，从而影响胚胎的长期发育。
Epidemiological investigations have found that air fine particulate matter (PM) exposure not only causes respiratory and cardiovascular diseases in adults and children, but also affects embryonic development during pregnancy, leading to poor pregnancy outcomes. However, its exact molecular mechanism is still unclear. In this study, human embryonic stem cells (hESCs) were treated with PM at different concentrations then the morphology and proliferation capacity were measured. The mRNA and protein expression of NANOG and OCT4 were detected using quantitative PCR, immunofluorescence, western blotting, and flow cytometry. Reactive oxygen species (ROS) generation and AKT/ERK activation were also measured. Meanwhile, changes in ROS, the expression of NANOG, OCT4, and the AKT/ERK pathways were measured in the hESCs with or without pretreatment of ROS scavenger N-acetylcysteine (NAC) prior to PM exposure. After PM exposure, the proliferation capacity and expression of OCT4 and NANOG at the mRNA and protein levels were downregulated. The ROS level in the hESCs increased after PM exposure, but this increase in ROS was attenuated by pretreatment with NAC. Further analysis showed that the levels of phosphorylated AKT and ERK increased after PM exposure. After pretreatment with NAC, the phosphorylation levels of AKT and ERK, which are crucial for regulating the proliferation, pluripotency, and differentiation of hESC, were significantly attenuated compared with the non-NAC pretreated exposure group. These results suggest that PM exposure may reduce the proliferation and pluripotency of hESC through ROS-mediated AKT/ERK pathways, thereby affecting the long-term development of embryos.