Polysaccharide from Fuzi Likely Protects Against Starvation-Induced Cytotoxicity in H9c2 Cells by Increasing Autophagy Through Activation of the AMPK/mTOR Pathway
Polysaccharide from Fuzi Likely Protects Against Starvation-Induced Cytotoxicity in H9c2 Cells by Increasing Autophagy Through Activation of the AMPK/mTOR Pathway
复制标题
附子多糖可能通过激活 AMPK/mTOR 通路来增加自噬，从而防止 H9c2 细胞中饥饿诱导的细胞毒性
DOI：
10.1142/s0192415x13500262
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发表时间：
2013-01-01
影响因子：
5.7
通讯作者：
Wu, Wei-Kang
中科院分区：
文献类型：
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作者：
Liao, Li-Zhen;Chen, Yan-Ling;Wu, Wei-Kang
有越来越多的证据表明，饥饿诱导自噬，这可能是保护饥饿期间，在AMPK依赖的方式。附子多糖（FPS）据报道对营养有限的肝脏具有保护作用。本研究旨在确定FPS是否使用AMPK/mTOR依赖性机制保护H9c2细胞免受饥饿诱导的细胞毒性。将H9c2细胞在血清和葡萄糖饥饿培养基中孵育12小时以建立细胞损伤模型。3-甲基腺嘌呤（3MA，一种自噬抑制剂）用于确定自噬在饥饿中的确切作用。将细胞与不同浓度的FPS孵育，并测量细胞损伤水平、自噬活性和AMPK/mTOR磷酸化。使用腺嘌呤9-β-D-阿拉伯呋喃糖苷（Ara-A，AMPK抑制剂）和5-氨基-4-咪唑甲酰胺核苷（AICAR，AMPK激活剂）来鉴定AMPK/mTOR通路是否参与FPS介导的心脏保护。我们证明了饥饿以时间依赖性的方式降低细胞活力，并且3MA诱导的自噬抑制加剧了细胞活力的降低。FPS处理减弱了细胞活力下降和饥饿诱导的线粒体膜电位（MMP）下降和自噬;此外，AMPK/mTOR通路在处理期间被激活。Ara-A处理可使FPS的保护作用消失，AICAR处理则与FPS作用相似。我们的结论是，自噬减弱饥饿诱导的心肌细胞死亡，FPS增加自噬活性，以防止饥饿诱导的细胞毒性H9c2细胞，可能通过AMPK/mTOR途径激活。
There is increasing evidence that starvation induces autophagy, which may be protective during starvation, in an AMPK-dependent manner. Polysaccharides from Fuzi (FPS) reportedly have protective effects on nutrition-limited livers. The present study was designed to determine whether FPS protected H9c2 cells against starvation-induced cytotoxicity using an AMPK/mTOR-dependent mechanism. H9c2 cells were incubated in serum and glucose starvation media for 12 hours to establish a cell injury model. 3-Methyladenine (3MA, an autophagy inhibitor) was used to identify the exact role of autophagy in starvation. Cells were incubated with different FPS concentrations, and the cell injury levels, autophagy activity and AMPK/mTOR phosphorylation were measured. Adenine 9-beta-D-arabinofuranoside (Ara-A, an AMPK inhibitor) and 5-amino-4-imidazole-carboxamide riboside (AICAR, an AMPK activator) were used to identify whether the AMPK/mTOR pathway was involved in FPS-mediated cardioprotection. We demonstrated that starvation decreased cell viability in a time-dependent manner, and 3MA-induced autophagy inhibition aggravated the reduced cell viability. FPS treatment attenuated the cell viability decrement and the starvation-induced decline in the mitochondrial membrane potential (MMP), and autophagy; also, the AMPK/mTOR pathways were activated during treatment. Ara-A treatment abolished the protective effect of FPS, while AICAR treatment had a similar effect to FPS. We conclude that autophagy attenuates starvation-induced cardiomyocyte death, and FPS increases autophagy activity to protect against starvation-induced cytotoxicity in H9c2 cells, likely through AMPK/mTOR pathway activation.