HT22细胞系铁死亡敏感性研究

doi:10.3969/j.issn.1006-7795.2020.01.017

首都医科大学学报 ›› 2020, Vol. 41 ›› Issue (1): 87-91.doi: 10.3969/j.issn.1006-7795.2020.01.017

HT22细胞系铁死亡敏感性研究

杨田丽^1,2, 杨勇飞³, 袁增强⁴

1. 首都医科大学脑重大疾病研究中心, 北京 100069;
2. 北京脑重大疾病研究院, 北京 100069;
3. 北京理工大学生命学院, 北京 100081;
4. 军事医学研究院军事认知与脑科学研究所, 北京 100850

收稿日期:2019-06-11 出版日期:2020-02-21 发布日期:2020-02-13
通讯作者: 袁增强 E-mail:zyuan620@yahoo.com
基金资助:
国家自然科学基金（31600946，81971091）。

Sensitivity study of HT22 in ferroptosis

Yang Tianli^1,2, Yang Yongfei³, Yuan Zengqiang⁴

1. Center for Brain Disorders Research, Capital Medical University, Beijing 100069, China;
2. Beijing Institute for Brain Disorders, Beijing 100069, China;
3. College of Lifescience, Beijing Institute of Technology, Beijing 100081, China;
4. The Brain Science Center, Beijing Institute of Basic Medical Sciences, Beijing 100850, China

Received:2019-06-11 Online:2020-02-21 Published:2020-02-13
Supported by:
This study was supported by National Natural Science Foundation of China(31600946,81971091).

摘要/Abstract

摘要： 目的检测HT22细胞系可否作为研究神经元铁死亡的良好细胞模型，在细胞水平上为研究神经元铁死亡的机制提供基础。方法用经典铁死亡诱导剂Erastin和RSL3处理HT22细胞系，显微镜下观察细胞存活情况；使用CCK-8法检测细胞存活率；使用丙二醛法或菲罗嗪法分别检测细胞内脂质活性氧（reactive oxygen species，ROS）、Fe²⁺浓度；实时定量聚合酶链反应（quantitative real time polymerase chain reaction，qRT-PCR）法检测细胞铁死亡关键基因SLC7A11（solute carrier family 7 member 11）、PTGS2（prostaglandin-endoperoxide synthase 2）的mRNA水平。结果 Erastin或RSL3处理HT22后，显微镜下发现细胞发生明显死亡，加入铁死亡抑制剂Ferrostatin-1后，细胞死亡显著减少。Erastin或RSL3诱导后HT22细胞内的Fe²⁺浓度无明显变化（P>0.05），脂质过氧化水平明显增加（P<0.05）；Erastin或RSL3导致细胞SLC7A11、PTGS2的mRNA表达水平增加（P<0.05），进一步验证了处理后细胞发生铁死亡。结论 HT22细胞系是一种铁死亡敏感细胞系，Erastin或RSL3诱导HT22铁死亡，HT22细胞系可用于研究神经元中铁死亡的发生及胞内脂质过氧化物蓄积机制。

关键词: 铁死亡, 神经元, 脂质过氧化, Fe²⁺

Abstract: Objectives To investigate whether HT22 cell line is a good cell model to study the ferroptosis and its molecular mechanisms. Methods HT22 cells were treated with Erastin or RSL3, the putative ferroptosis inducers, followed by the observation of morphology change under microscope. Moreover, cell viability was measured by cell counting kit 8 (CCK-8), and cellular lipid peroxidation was measured by MDA assay and the cellular concentration of Fe²⁺ was measured by Fe assay. In addition, the mRNA expression levels of SLC7A11 and PTGS2 were measured by quantitative real time polymerase chain reaction(qRT-PCR). Results Erastin or RSL3 treatment significantly increased lipid peroxidation in HT22, which lead to ferroptosis. However, Erastin or RSL3 did not alter the Fe²⁺ concentration in HT22 cells. In addition, we found that Erastin or RSL3 transcriptionally upregulated the expressions of SLC7A11 and PTGS2. Conclusion HT22 is a suitable cell model to study ferroptosis and the underlying mechanism of the accumulation of lipid peroxidation during ferroptosis.

Key words: ferroptosis, neurons, lipid peroxidation, Fe²⁺

中图分类号:

Q189

杨田丽, 杨勇飞, 袁增强. HT22细胞系铁死亡敏感性研究[J]. 首都医科大学学报, 2020, 41(1): 87-91.

Yang Tianli, Yang Yongfei, Yuan Zengqiang. Sensitivity study of HT22 in ferroptosis[J]. Journal of Capital Medical University, 2020, 41(1): 87-91.

参考文献

[1] Dixon S J,Lemberg K M,Lamprecht M R,et al. Ferroptosis:an iron-dependent form of nonapoptotic cell death[J]. Cell,2012,149(5):1060-1072.
[2] Yagoda N,von Rechenberg M,Zaganjor E,et al. RAS-RAF-MEK-dependent oxidative cell death involving voltage-dependent anion channels[J]. Nature,2007,447(7146):864-868.
[3] Ye F, Chai W, Xie M, et al. HMGB1 regulates erastin-induced ferroptosis via RAS-JNK/p38 signaling in HL-60/NRAS (Q61L) cells[J]. Am J Cancer Res, 2019, 9(4):730-739.
[4] Li L B, Chai R, Zhang S, et al. Iron exposure and the cellular mechanisms linked to neuron degeneration in adult mice[J]. Cells, 2019, 8(2):198-215.
[5] Matsushita M, Freigang S, Schneider C, et al. T cell lipid peroxidation induces ferroptosis and prevents immunity to infection[J]. J Exp Med, 2015, 212(4):555-568.
[6] Friedmann Angeli J P,Schneider M,Proneth B,et al. Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice[J]. Nat Cell Biol,2014,16(12):1180-1191.
[7] Adedoyin O, Boddu R, Traylor A, et al. Heme oxygenase-1 mitigates ferroptosis in renal proximal tubule cells[J]. Am J Physiol Renal Physiol, 2018, 314(5):702-714.
[8] 王镜岩,朱圣庚,徐长法,等.生物化学(上册)[M]. 3版.北京:高等教育出版社,2002:96-103.
[9] Sui X, Zhang R, Liu S, et al. RSL3 drives ferroptosis through GPX4 inactivation and ROS production in colorectal cancer[J]. Front Pharmacol, 2018, 9(1):1371-1388.
[10] Wang C, Cai X, Hu W, et al. Investigation of the neuroprotective effects of crocin via antioxidant activities in HT22 cells and in mice with Alzheimer's disease[J]. Int J Mol Med, 2019, 43(2):956-966.
[11] Li Z, Chen X, Lu W, et al. Anti-oxidative stress activity is essential for amanita caesarea mediated neuroprotection on glutamate-induced apoptotic HT22 cells and an alzheimer's disease mouse model[J]. Int J Mol Sci, 2017, 18(8):1623-1636.
[12] Lee D S, Cheong S H. Taurine have neuroprotective activity against oxidative damage-induced HT22 death through heme oxygenase-1 pathway[J]. Adv Exp Med Biol, 2017, 975(Pt 1):159-171.
[13] Yang W S, Stockwell B R. Ferroptosis:death by lipid peroxidation[J]. Trends Cell Biol,2016,26(3):165-176.
[14] Zhang Z, Wu Y, Yuan S, et al. Glutathione peroxidase 4 participates in secondary brain injury through mediating ferroptosis in a rat model of intracerebral hemorrhage[J]. Brain Res, 2018, 1701(1):112-125.
[15] Li Q, Han X, Lan X, et al. Inhibition of neuronal ferroptosis protects hemorrhagic brain[J]. JCI Insight, 2017, 2(7):90777-90796.
[16] Fine J M, Forsberg A C, Stroebel B M, et al. Intranasal deferoxamine affects memory loss, oxidation, and the insulin pathway in the streptozotocin rat model of Alzheimer's disease[J]. J Neurol Sci, 2017, 380(1):164-171.

HT22细胞系铁死亡敏感性研究

Sensitivity study of HT22 in ferroptosis

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