Dexamethasone-induced production of reactive oxygen species promotes apoptosis via endoplasmic reticulum stress and autophagy in MC3T3-E1 cells.
Dexamethasone-induced production of reactive oxygen species promotes apoptosis via endoplasmic reticulum stress and autophagy in MC3T3-E1 cells.
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地塞米松诱导的活性氧产生通过内质网应激和自噬促进 MC3T3-E1 细胞凋亡
DOI：
10.3892/ijmm.2018.3412
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发表时间：
2018-04
影响因子：
5.4
通讯作者：
Bai R
中科院分区：
文献类型：
--
作者：
Liu W;Zhao Z;Na Y;Meng C;Wang J;Bai R
长期或过量使用糖皮质激素（GC）引发的成骨细胞凋亡已被确定为骨质疏松症和骨坏死发展的主要贡献者。然而，GC诱导细胞凋亡的分子机制是多方面的，仍有待充分阐明。本研究旨在探讨地塞米松（DEX）诱导的MC 3 T3-E1成骨样细胞活性氧（ROS）、自噬和凋亡之间的关系。使用细胞计数试剂盒-8测定法评估细胞活力，并进行流式细胞术以评估细胞凋亡、细胞周期和ROS产生。免疫荧光和Western blot分别检测自噬泡和细胞周期蛋白D激酶（CDK）2、多聚腺苷二磷酸核糖聚合酶（poly[ADP ribose] polymerase）、半胱氨酸天冬氨酸蛋白酶（caspase-3）、转录激活因子（ATF）4、CCAAT/增强子结合蛋白同源蛋白（CHOP）、Beclin 1、微管相关蛋白1A/1B轻链（LC）3B和P62蛋白的表达。结果表明，DEX不仅降低细胞活力，而且通过激活内质网（ER）应激促进细胞凋亡。DEX通过抑制CDK 2的表达，使细胞周期阻滞于G 0/G1期，并激活ROS的产生。值得注意的是，在用3-甲基腺嘌呤（自噬抑制剂）处理后，DEX介导的存活力和凋亡的变化在MC 3 T3-E1细胞中减弱。用抗氧化剂N-乙酰半胱氨酸处理可消除DEX对MC 3 T3-E1细胞增殖、凋亡、ER应激和自噬的影响。总之，本研究结果表明，DEX促进了ROS的产生，从而通过激活自噬和ER应激促进了MC 3 T3-E1细胞的凋亡。
Apoptosis of osteoblasts, triggered by prolonged or excessive use of glucocorticoids (GCs), has been identified as a dominant contributor to the development of osteoporosis and osteonecrosis. However, the molecular mechanisms underlying GC-induced apoptosis are multifaceted and remain to be fully elucidated. The present study aimed to explore the correlation between dexamethasone (DEX)-induced reactive oxygen species (ROS), autophagy and apoptosis in MC3T3-E1 osteoblast-like cells. Cell viability was assessed using a Cell Counting Kit-8 assay, and flow cytometry was performed to assess cellular apoptosis, cell cycle and ROS production. Immunofluorescence and western blot analysis were respectively used to detect autophagic vacuoles and the expression of proteins, including cyclin D kinase (CDK)2, poly[ADP ribose] polymerase, caspase-3, activating transcription factor (ATF)4, CCAAT/enhancer-binding protein homologous protein (CHOP), Beclin1, microtubule-associated proteins 1A/1B light chain (LC)3B and P62. It was revealed that DEX not only reduced cell viability, but also promoted apoptosis via the activation of endoplasmic reticulum (ER) stress. In addition, DEX induced cell cycle arrest at G0/G1 phase via inhibition of the expression of CDK2, and the production of ROS was activated. Of note, the DEX-mediated changes in viability and apoptosis were attenuated in MC3T3-E1 cells after treatment with 3-methyladenine, which is an autophagy inhibitor. Treatment with the antioxidant N-acetylcysteine abolished the effect of DEX on the proliferation, apoptosis, ER stress and autophagy of MC3T3-E1 cells. In conclusion, the present results indicated that DEX promoted the production of ROS, which enhanced apoptosis through activation of autophagy and ER stress in MC3T3-E1 cells.
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DOI：
10.1038/nrm3735
发表时间：
2014-02
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1007/978-1-60761-411-1_4
发表时间：
2010-01-01
期刊：
ADVANCED PROTOCOLS IN OXIDATIVE STRESS II
影响因子：
--
作者：
Eruslanov, Evgeniy;Kusmartsev, Sergei
通讯作者：
Kusmartsev, Sergei
DOI：
10.2147/dddt.s65410
发表时间：
2014
期刊：
Drug design, development and therapy
影响因子：
--
作者：
Lin H;Wei B;Li G;Zheng J;Sun J;Chu J;Zeng R;Niu Y
通讯作者：
Niu Y
影响因子：
7.2
作者：
Gu, G;Hentunen, TA;Väänänen, HK
通讯作者：
Väänänen, HK
DOI：
10.1196/annals.1391.007
发表时间：
2007-01-01
期刊：
STRESS RESPONSES IN BIOLOGY AND MEDICINE
影响因子：
--
作者：
Banhegyi, Gabor;Baumeister, Peter;Yang, Wei
通讯作者：
Yang, Wei