腺苷酸活化蛋白激酶对缺氧肾小管上皮细胞线粒体片段化的影响
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摘要
目的:探讨腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)缺氧环境中对肾小管上皮细胞线粒体片段化的影响。方法:采用AnaeroPack System建立缺氧模型,在缺氧前加入Compound C抑制AMPK活性。缺氧培养肾小管上皮细胞48小时后,蛋白免疫印迹法检测AMPK磷酸化水平,并用线粒体示踪染料标记线粒体后在共聚焦显微镜下观察线粒体形态,并统计线粒体片段化细胞的数目及比例。结果:缺氧后肾小管上皮细胞线粒体片段化明显,伴有AMPK磷酸化水平显著升高。予以Compound C抑制AMPK活性后,肾小管上皮细胞线粒体片段化显著减少。结论:缺氧肾小管上皮细胞的线粒体片段化与AMPK密切相关,减少AMPK磷酸化可减轻缺氧肾小管上皮细胞线粒体片段化,保护细胞。
Objective: To investigate the role of AMP-activated protein kinase on mitochondrial fragmentation of renal tubular epithelial cells(RTECs)under hypoxia. Methods:RTPECs were cultured in the AnaeroPack system to induce hypoxia. Compound C was added to inhibit AMPK activation before hypoxia. Cells were stained with the mitochondrial dye and themorphology of mitochondria was observed by confocal microscope after 48 hours. The proportion of cells which underwent mitochondrial fragmentation was calculated. The level of AMPK phosphorylation was detected by Western blot. Results:When subjected to hypoxia cultivation for 48 hours,mitochondria of RTECs underwent excessive fragmentation and AMPK was greatly activated. After pretreated with Compound C,mitochondrial fragmentation was significantly alleviated. Conclusion:Mitochondrial fragmentation of RTECs under hypoxia was closely related to AMPK activation. Inhibition of AMPK could alleviate mitochondrial fragmentation and protect RTECs.
Objective: To investigate the role of AMP-activated protein kinase on mitochondrial fragmentation of renal tubular epithelial cells(RTECs)under hypoxia. Methods:RTPECs were cultured in the AnaeroPack system to induce hypoxia. Compound C was added to inhibit AMPK activation before hypoxia. Cells were stained with the mitochondrial dye and themorphology of mitochondria was observed by confocal microscope after 48 hours. The proportion of cells which underwent mitochondrial fragmentation was calculated. The level of AMPK phosphorylation was detected by Western blot. Results:When subjected to hypoxia cultivation for 48 hours,mitochondria of RTECs underwent excessive fragmentation and AMPK was greatly activated. After pretreated with Compound C,mitochondrial fragmentation was significantly alleviated. Conclusion:Mitochondrial fragmentation of RTECs under hypoxia was closely related to AMPK activation. Inhibition of AMPK could alleviate mitochondrial fragmentation and protect RTECs.
引文
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[2] Zhan M,Brooks C,Liu F,et al. Mitochondrial dynamics:regulatory mechanisms and emerging role in renal pathophysiology[J]. Kidney Int,2013,83(4):568.
[3] Parikh SM,Yang Y,He L,et al. Mitochondrial function and disturbances in the septic kidney[J]. Semin Nephrol,2015,35(1):108.
[4] Bhargava P,Schnellmann RG. Mitochondrial energetics in the kidney[J]. Nat Rev Nephrol,2017,13(10):629.
[5] Pei S,MinhajuddinM,Adane B,et al. AMPK/FIS1-Mediated Mitophagy Is Required for Self-Renewal of Human AML Stem Cells[J]. Cell Stem Cell,2018,23(1):86.
[6] Ducommun S,Deak M,Sumpton D,et al. Motif affinity and mass spectrometry proteomic approach for the discovery of cellular AMPK targets:identification of mitochondrial fission factor as a new AMPK substrate[J]. Cell Signal,2015,27(5):978.
[7] Toyama EQ,HerzigS,Courchet J,et al. AMP-activated protein kinase mediates mitochondrial fission in response to energy stress[J]. Science,2016,351(6270):275.
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