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首页> 外文期刊>Cellular Physiology and Biochemistry >Inhibition of the Na+-H+ exchanger isoform-1 and the extracellular signal-regulated kinase induces apoptosis: A time course of events
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Inhibition of the Na+-H+ exchanger isoform-1 and the extracellular signal-regulated kinase induces apoptosis: A time course of events

机译：Na +-H +交换异构体1和细胞外信号调节激酶的抑制诱导细胞凋亡：事件的时间进程。

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Aims: The present study attempts to shed light on the role and the relative position of the Na+/H+ exchanger isoform 1 (NHE1) and the extracellular signal-regulated kinase (ERK) in HEp-2 cell signaling pathways concerning a diverse range of cellular functions such as regulation of intracellular pH (pHi), DNA synthesis, production of reactive oxygen species (ROS) and apoptosis. Methods: Pharmacological inhibition with cariporide (highly specific inhibitor of NHE1) and PD98059 (specific inhibitor of the upstream activator of ERK) was implemented. Fluorescence spectrometry, atomic absorption spectrometry and ELISA methods were used in order to obtain the results. Results: NHE1 and ERK take part in all of the aforementioned cellular functions, as their inhibition had an effect on all of them. Additionally, inhibition of NHE1 resulted in ERK inhibition as well. Moreover, continuous inhibition of NHE1 or ERK for up to 24h led HEp-2 cells to apoptosis, as assessed through caspase-3 activation, DNA fragmentation and annexin-V binding levels. Conclusion: Our data shows a time course of events in relation to NHE1 and ERK and suggests the existence of a positive feedback loop between NHE1 and ERK which could pose a barrier against apoptosis.
机译：目的：本研究试图阐明Na + / H +交换异构体1（NHE1）和细胞外信号调节激酶（ERK）在HEp-2细胞信号通路中的作用和相对位置，涉及多种细胞这些功能包括调节细胞内pH（pHi），DNA合成，产生活性氧（ROS）和凋亡。方法：使用卡立哌肽（NHE1的高度特异性抑制剂）和PD98059（ERK上游激活剂的特异性抑制剂）进行药理抑制。为了得到结果，使用了荧光光谱法，原子吸收光谱法和ELISA法。结果：NHE1和ERK参与了所有上述细胞功能，因为它们的抑制作用都对它们都有影响。此外，NHE1的抑制也导致ERK抑制。此外，通过caspase-3活化，DNA片段化和膜联蛋白-V结合水平评估，持续抑制NHE1或ERK长达24小时导致HEp-2细胞凋亡。结论：我们的数据显示了与NHE1和ERK有关的事件的时间过程，并表明NHE1和ERK之间存在正反馈回路，这可能对细胞凋亡构成障碍。

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