Sestrin2通过Nrf2/xCT途径减轻七氟醚诱导的HT22细胞铁死亡研究

李福军; 姜涛; 郭力甲; 王顺

doi:10.16766/j.cnki.issn.1674-4152.001949

Sestrin2通过Nrf2/xCT途径减轻七氟醚诱导的HT22细胞铁死亡研究

doi: 10.16766/j.cnki.issn.1674-4152.001949

李福军^{1, 2},
姜涛³,
郭力甲²,
王顺^1, ,

1.
黑龙江省中医药科学院针灸科，黑龙江哈尔滨 150036
2.
哈尔滨医科大学附属第一医院麻醉科，黑龙江哈尔滨 150036
3.
哈尔滨医科大学附属第二医院麻醉科，黑龙江哈尔滨 150036

基金项目:

黑龙江省卫生计生委科学研究基金项目 2018318

详细信息

通讯作者:
王顺，E-mail: hljwang@yahoo.com.cn

中图分类号: R971.2 R329.28
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- 文章访问数: 161
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- PDF下载量: 3
- 被引次数: 0
出版历程
- 收稿日期: 2021-01-29
- 网络出版日期: 2022-02-16

Sestrin 2 extenuates neuronal ferroptosis induced by sevoflurane through the Nrf2/xCT pathway

LI Fu-jun^{1, 2},
JIANG Tao³,
GUO Li-jia²,
WANG Shun^{1
, ,}

1.
Acupuncture and Moxibustion Department, Heilongjiang College of Traditional Chinese Medicine, Harbin, Heilongjiang 150036, China

摘要

摘要: 目的探讨Sestrin 2(SESN2)在七氟醚(SEV)诱导神经元损伤的作用及其分子调控机制。方法将HT22小鼠海马神经元细胞用0%、1%、2%、4%七氟醚混合气体和/或铁死亡抑制剂Ferrostatin-1(FER-1)处理6 h。将SESN2过表达载体(SESN2-OE)和相应的对照(NC)转染至HT22细胞中。检测细胞活力及铁离子、活性氧(ROS)、丙二醛(MDA)和总谷胱甘肽(GSH)水平。实时定量PCR(qPCR)和Western blotting测定mRNA和蛋白水平。结果 SEV明显抑制SESN2 mRNA和蛋白的表达(t值分别为22.904、37.432，均P < 0.05)，转染SESN2-OE后SESN2 mRNA和蛋白表达明显增加(t值分别为12.703、11.687，均P < 0.01)。SEV处理使HT22细胞中铁离子、ROS和MDA的水平明显增加(t值分别为29.031、18.819、28.054, 均P < 0.05)，而GSH、Nrf2和胱氨酸/谷氨酸逆向转运体(xCT)的水平明显降低(t值分别为14.617、34.513、13.836，均P < 0.05)。转染SESN2-OE使SEV处理的HT22细胞中铁离子、ROS和MDA的水平明显降低(t值分别为8.342、9.373、7.381，均P < 0.05)，而GSH、核因子E2相关因子(Nrf2)和xCT的水平明显增加(t值分别为12.718、10.102、9.814，均P < 0.05)。结论 Sestrin 2通过激活Nrf2/xCT途径抑制七氟醚诱导的HT22神经元铁死亡。
- Sestrin 2 /
- 七氟醚 /
- 铁死亡 /
- 神经元损伤
Abstract: Objective To explore the role of Sestrin 2 (SESN2) in neuronal ferroptosis induced by sevoflurane (SEV) and its possible mechanism. Methods Mouse hippocampal neuron cells HT22 were treated with 0%, 1%, 2% or 4% sevoflurane and iron death inhibitor Ferrostatin-1 (FER-1) for 6 h. SESN2 overexpression vector (SESN2-OE) and corresponding control (NC) were transfected into HT22 cells. The CCK-8 assay was used in detecting cell viability, and the corresponding kit was used in detecting the levels of iron, reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH). The expression levels of genes and proteins were detected through real-time quantitative PCR (qPCR) and western blot assay. Results qPCR and western blot results showed that SEV treatment significantly inhibited the expression of SESN2 mRNA and protein (t=22.904, 37.432; P < 0.05), whereas transfection with SESN2-OE significantly increased the expression levels of SESN2 mRNA and protein in SEV-treated HT22 cells (t=12.703, 11.687; P < 0.05). SEV treatment significantly increased the levels of iron, ROS and MDA in the HT22 cells (t=29.031, 18.819, 28.054) but significantly reduced the levels of GSH, Nrf2 and xCT (t=14.617, 34.513, 13.836; all P < 0.05). Transfection with SESN2-OE significantly reduced the levels of iron, ROS and MDA in SEV-treated HT22 cells (t=8.342, 9.373, 7.381) but significantly increased the levels of GSH, Nrf2 and xCT (t=12.718, 10.102, 9.814; P < 0.05). Conclusion Sestrin 2 inhibited sevoflurane-induced HT22 neuronal ferroptosis by activating the Nrf2/xCT pathway.
- Sestrin 2 /
- Sevoflurane /
- Ferroptosis /
- Neuronal damage

HTML全文

图 1 HT22细胞中SESN2 mRNA和蛋白的表达

注：A为qPCR检测SESN2 mRNA的表达；B为Western blotting检测SENS2蛋白的表达；C为SENS2蛋白相对表达水平；与对照组比较，^aP＜0.001；与SEV+NC组比较，^bP＜0.01。

下载: 全尺寸图片幻灯片

图 2 HT22细胞中Nrf2和xCT蛋白的表达

注：与对照组比较，^aP＜0.01；与SEV+NC组比较，^bP＜0.01。

下载: 全尺寸图片幻灯片

参考文献(25)

[1]	王加仑, 廉海光, 林凤颜, 等. 新生小鼠七氟醚暴露对其青春期记忆及突触可塑性的影响[J]. 安徽医科大学学报, 2020, 55(4): 513-517. https://www.cnki.com.cn/Article/CJFDTOTAL-YIKE202004006.htm
[2]	WU Y, SONG J, WANG Y, et al. The Potential Role of ferroptosis in neonatal brain injury[J]. Front Neurosci, 2019, 13(14): 115.
[3]	JIN H R, DU C H, WANG C Z, et al. Ginseng metabolite protopanaxadiol induces Sestrin 2 expression and AMPK activation through GCN2 and PERK[J]. Cell Death Dis, 2019, 10(4): 311. doi: 10.1038/s41419-019-1548-7
[4]	HAN X, DING C H, ZHANG G D, et al. Liraglutide ameliorates obesity-related nonalcoholic fatty liver disease by regulating Sestrin 2-mediated Nrf2/HO-1 pathway[J]. Biochem Biophys Res Commun, 2020, 525(4): 895-901. doi: 10.1016/j.bbrc.2020.03.032
[5]	LI Y X, WU J X, YU S S, et al. Sestrin 2 promotes angiogenesis to alleviate brain injury by activating Nrf2 through regulating the interaction between p62 and Keap1 following photothrombotic stroke in rats[J]. Brain Res, 2020, 1745(15): 146948. http://www.sciencedirect.com/science/article/pii/S0006899320303048
[6]	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. doi: 10.3969/j.issn.1006-7795.2020.01.017
[7]	鲁瑞涛. 丁苯酞对七氟醚诱导神经细胞损伤的保护作用[J]. 中华细胞与干细胞杂志(电子版), 2018, 8(1): 12-16. doi: 10.3877/cma.j.issn.2095-1221.2018.01.003
[8]	沈亚建, 方军, 周惠丹, 等. 七氟醚和丙泊酚对行腹腔镜宫颈癌根治术患者炎症因子及应激反应的影响[J]. 中华全科医学, 2017, 15(1): 66-68. https://www.cnki.com.cn/Article/CJFDTOTAL-SYQY201701022.htm
[9]	梁艳宁, 周伟. 七氟醚吸入麻醉对急性颅内出血患者呼吸循环系统及脑功能的影响[J]. 医药论坛杂志, 2021, 42(1): 13-16, 21. https://www.cnki.com.cn/Article/CJFDTOTAL-HYYX202101004.htm
[10]	温娟, 欧刚, 宋振鹏, 等. 不同浓度七氟醚对肝脏清除功能和血流动力学的影响[J]. 临床麻醉学杂志, 2020, 36(3): 266-270. https://www.cnki.com.cn/Article/CJFDTOTAL-LCMZ202003015.htm
[11]	尹彩星, 龚清安, 司小萌. 不同浓度七氟醚对急性心肌缺血再灌注老龄大鼠血清、心脏5-HT、白细胞介素和海马组织SOD、MDA的影响[J]. 药物评价研究, 2020, 43(4): 700-705. https://www.cnki.com.cn/Article/CJFDTOTAL-YWPJ202004020.htm
[12]	RAPER J, DE BIASIO J C, MURPHY K L, et al. Persistent alteration in behavioural reactivity to a mild social stressor in rhesus monkeys repeatedly exposed to sevoflurane in infancy[J]. Br J Anaesth, 2018, 120(4): 761-767. doi: 10.1016/j.bja.2018.01.014
[13]	KANG W, LU D, YANG X, et al. Sevoflurane induces hippocampal neuronal apoptosis by altering the level of neuropeptide Y in neonatal rats[J]. Neurochem Res, 2020, 45(9): 1986-1996. doi: 10.1007/s11064-020-03028-9
[14]	ZHANG N N, WANG D, YANG X L, et al. Long noncoding RNA small nucleolar RNA host gene 1 contributes to sevoflurane-induced neurotoxicity through negatively modulating microRNA-181b[J]. Neuroreport, 2020, 31(5): 416-424. doi: 10.1097/WNR.0000000000001430
[15]	魏元辉, 桂燕妮, 姜淑娟. 铁死亡在肺癌中的作用研究进展[J]. 中华医学杂志, 2019, 99(10): 796-798.
[16]	ANGELI J P F, SHAH R, PRATT D A, et al. Ferroptosis inhibition: mechanisms and opportunities[J]. Trends Pharmacol Sci, 2017, 38(5): 489-498. doi: 10.1016/j.tips.2017.02.005
[17]	LEI P X, BAI T, SUN Y L. Mechanisms of ferroptosis and relations with regulated cell death: a review[J]. Front Physiol, 2019, 10(26): 139. http://www.ncbi.nlm.nih.gov/pubmed/30863316
[18]	FANG F, XUE Z, CANG J. Sevoflurane exposure in 7-day-old rats affects neurogenesis, neurodegeneration and neurocognitive function[J]. Neurosci Bull, 2012, 28(5): 499-508. doi: 10.1007/s12264-012-1260-4
[19]	LV X, YAN J, JIANG J, et al. MicroRNA-27a-3p suppression of peroxisome proliferator-activated receptor-γ contributes to cognitive impairments resulting from sevoflurane treatment[J]. J Neurochem, 2017, 143(3): 306-319. doi: 10.1111/jnc.14208
[20]	王雪冬, 王迎迎, 刘索宁, 等. 七氟醚对HT22小鼠海马神经元细胞DNA的损伤及其机制[J]. 吉林大学学报(医学版), 2020, 46(2): 240-247, 432. https://www.cnki.com.cn/Article/CJFDTOTAL-BQEB202002006.htm
[21]	PARK S J, CHO S S, KIM K M, et al. Protective effect of Sestrin 2 against iron overload and ferroptosis-induced liver injury[J]. Toxicol Appl Pharmacol, 2019, 379(15): 114665. http://www.ncbi.nlm.nih.gov/pubmed/31323261
[22]	VIVARINI A C, LOPES U G. The Potential Role of Nrf2 signaling in leishmania infection outcomes[J]. Front Cell Infect Microbiol, 2019, 9(10): 453. http://www.ncbi.nlm.nih.gov/pubmed/31998662
[23]	ABDALKADER M, LAMPINEN R, KANNINEN K M, et al. Targeting Nrf2 to suppress ferroptosis and mitochondrial dysfunction in neurodegeneration[J]. Front Neurosci, 2018, 12(10): 466. http://www.onacademic.com/detail/journal_1000040522356510_ad0f.html
[24]	LI Y, YAN H, XU X M, et al. Erastin/sorafenib induces cisplatin-resistant non-small cell lung cancer cell ferroptosis through inhibition of the Nrf2/xCT pathway[J]. Oncol lett, 2020, 19(1): 323-333. http://www.ingentaconnect.com/content/sp/ol/2020/00000019/00000001/art00036
[25]	YANG X X, YANG H B, WU F D, et al. Mn inhibits GSH synthesis via downregulation of neuronal EAAC1 and astrocytic xCT to cause oxidative damage in the striatum of mice[J]. Oxid Med Cel Longev, 2018, 2018(30): 4235695. http://www.onacademic.com/detail/journal_1000040912762210_f557.html