磷酸腺苷激活的蛋白激酶(AMPK)参与了曲格列酮诱导的HeLa细胞自噬和凋亡

AMP-activated protein kinase-α is involved in the autophagy and apoptosis caused by troglitazone

摘要:

摘要:【目的】明确磷酸腺苷激活的蛋白激酶(AMPK)在细胞自噬和凋亡中的作用。【方法】利用电镜、荧光显微镜、蛋白免疫杂交、siRNA 干扰、流式细胞计数、MTS 细胞活性检测等对曲格列酮(troglitazone，TZ)处理的HeLa 细胞自噬和凋亡情况进行了检测。【结果】不同检测方法均表明TZ 增加了HeLa 细胞的自噬，这种自噬的发生伴随着AMPK 的磷酸化的降低;抑制AMPK 增加基础细胞自噬，而阻断了TZ 引起的自噬标记物LC3-II 的增加，同时也减少了TZ 引起的凋亡分子PARP 的切割;用自噬抑制剂3-MA 和干扰细胞自噬基因，不仅PARP 的切割明显地受到抑制，而且也部分阻断了TZ 引起的细胞活性丧失。【结论】AMPK 直接参与了TZ 引起的HeLa 细胞自噬过程，这种自噬发生促进了其诱导的细胞凋亡。

关键词:关键词:AMPK，自噬，凋亡，曲格列酮

Abstract:

Abstract:［Objective］To reveal the mechanism of AMPK signaling in the autophagy and apoptosis caused by troglitazone (TZ)．［Methods］To investigate the effect of TZ on alteration of autophagy and apoptosis in HeLa cells，fluorescence microscopy，electron microscopy，western-blotting，siRNA interference，flow cytometry and MTS assay were used．［Results］TZ attenuated AMP-activated protein kinase-α (AMPKα) phosphorylation，and stimulates autophagic process in HeLa cells．TZ induced the accumulation of microtubule-associated protein 1 light chain 3-II (LC3-II)，and degradation of sequestosome 1 (SQSTM1/p62)，two markers of autophagy，occurring prior to the caspase activation．Compound C，an AMPK inhibitor，increased basal and inhibits TZ-stimulated LC3-II content and TZ-depended PARP cleavage． Knockdown of the gene encoding autophagic proteins and AMPK conferred the cells resistance to apoptosis by TZ．［Conclusion］ Taken together，these data demonstrate that AMPK is involved in TZ promote autophagy，which precedes and contributes to caspase-dependent apoptosis．

Key words:Keywords: AMPK，Autophagy，Apoptosis，Troglitazone