Ferrostatin-1通过抑制铁死亡延缓D-gal诱导的心肌细胞衰老的研究

doi:10.11958/20220818

天津医药 ›› 2023, Vol. 51 ›› Issue (1): 19-23.doi: 10.11958/20220818

Ferrostatin-1通过抑制铁死亡延缓D-gal诱导的心肌细胞衰老的研究

熊喜成¹(), 王一平¹, 王刚¹, 张甜¹, 包雅丽¹, 迪娜·艾尼瓦尔¹, 孙湛¹^,²^,^∆()

1 新疆医科大学基础医学院病理生理学教研室（邮编830000）
2 新疆地方病分子生物学重点实验室

收稿日期:2022-05-24 修回日期:2022-07-12 出版日期:2023-01-15 发布日期:2023-01-17
通讯作者: 孙湛 E-mail:2111697911@qq.com;sunzhan724@126.com
作者简介:熊喜成（1994），男，硕士在读，主要从事心血管疾病基础研究。E-mail：2111697911@qq.com
基金资助:
新疆维吾尔自治区自然科学基金资助项目(2019D01C207);新疆地方与民族高发病教育部重点实验室开放基金(KF2021-5)

Ferrostatin-1 delay D-gal induced cardiomyocyte senescence by inhibiting ferroptosis

XIONG Xicheng¹(), WANG Yiping¹, WANG Gang¹, ZHANG Tian¹, BAO Yali¹, Di na·AINIWAER¹, SUN Zhan¹^,²^,^∆()

1 Department of Pathophysiology, College of Basic Medicine, Xinjiang Medical University, Urumqi 830000, China
2 Xinjiang Key Laboratory of Molecular Biology for Endemic Diseases

Received:2022-05-24 Revised:2022-07-12 Published:2023-01-15 Online:2023-01-17
Contact: SUN Zhan E-mail:2111697911@qq.com;sunzhan724@126.com

熊喜成, 王一平, 王刚, 张甜, 包雅丽, 迪娜·艾尼瓦尔, 孙湛. Ferrostatin-1通过抑制铁死亡延缓D-gal诱导的心肌细胞衰老的研究[J]. 天津医药, 2023, 51(1): 19-23.

XIONG Xicheng, WANG Yiping, WANG Gang, ZHANG Tian, BAO Yali, Di na·AINIWAER, SUN Zhan. Ferrostatin-1 delay D-gal induced cardiomyocyte senescence by inhibiting ferroptosis[J]. Tianjin Medical Journal, 2023, 51(1): 19-23.

摘要/Abstract

摘要：

目的探讨D-半乳糖（D-gal）诱导的心肌细胞衰老是否存在铁死亡及铁死亡抑制剂Ferrostatin-1（Fer-1）能否延缓心肌细胞衰老。方法通过不同水平（0、5、10、20、40、80、100 g/L）D-gal诱导H9C2心肌细胞损伤从而制备心脏衰老模型，采用MTT法检测细胞活力，确定后续实验采用的D-gal质量浓度。将细胞分为Control组、D-gal组和Fer-1组。MTT法检测细胞活力;DCFH-DA法检测细胞内活性氧（ROS）水平;微板法检测细胞内还原型谷胱甘肽（GSH）含量;硫代巴比妥酸法检测细胞内丙二醛（MDA）含量;Western blot检测溶质载体家族7成员11（SLC7A11）、谷胱甘肽过氧化物酶4（GPx4）、P53蛋白表达水平;微量法检测细胞内β-半乳糖苷酶（β-GAL）活性。结果 MTT法结果显示，细胞活力随D-gal质量浓度升高而降低（P<0.05），后续实验采用D-gal质量浓度为20 g/L。与D-gal组相比，Fer-1组细胞活力增加（P<0.05）;与Control组相比，D-gal组中ROS、MDA含量、P53蛋白表达水平、β-GAL活性增强，GSH含量、SLC7A11、GPx4蛋白表达水平降低（P<0.05）;与D-gal组相比，Fer-1组中ROS、MDA含量、P53蛋白表达水平、β-GAL活性降低，GSH、SLC7A11、GPx4蛋白表达升高（P<0.05）。结论 D-gal诱导的心肌细胞衰老过程中发生了铁死亡，Ferrostatin-1可抑制铁死亡，从而延缓心肌细胞衰老。

关键词: 肌细胞，心脏, 细胞衰老, 半乳糖, 铁死亡, D-gal, Ferrostatin-1

Abstract:

Objective To investigate the existence of ferroptosis in cardiomyocyte senescence induced by D-galactose (D-gal) and whether ferroptosis inhibitor ferrostatin-1 (Fer-1) can delay cardiomyocyte senescence. Methods Cardiomyocyte injury was induced by D-gal (0, 5, 10, 20, 40, 80 and 100 g/L) to simulate cardiac aging model. After cells were treated with different concentrations of D-gal for 24 hours, the cell viability was detected by MTT method to determine the concentration of D-gal in the follow-up experiment. The cells were divided into the control group, the D-gal group and the Fer-1 group. The cell viability was detected by MTT method. The level of intracellular reactive oxygen species (ROS) was detected by DCFH-DA method. The intracellular glutathione (GSH) content was detected by microplate method. Intracellular malondialdehyde (MDA) content was detected by thiobarbituric acid method. The protein expression levels of SLC7A11, GPx4 and P53 were detected by Western blot assay. The activity of intracellular β-galactosidase (β-GAL) was detected by microassay. Results MTT assay showed that the cell viability decreased with the increase of D-gal concentration (P<0.05). The concentration of D-gal was 20 g/L in subsequent experiments. Compared with the D-gal group, the cell viability was increased in the Fer-1 group (P<0.05). Compared with the control group, ROS and MDA contents, P53 protein expression level and β -GAL activity were increased in the D-gal group, while GSH content, SLC7A11 and GPx4 protein expression levels were decreased (P<0.05). Compared with the D-gal group, ROS and MDA contents, P53 protein expression level and β -GAL activity were decreased in the Fer-1 group, while GSH, SLC7A11 and GPx4 protein expressions were increased (P<0.05). Conclusion Ferroptosis occurs during D-gal-induced cell senescence, and Fer-1 can inhibit iron death and delay myocardial senescence.

Key words: myocytes, cardiac, cell aging, galactose, ferroptosis, D-gal, Ferrostatin-1

中图分类号:

R363.21

图/表 4

图1 D-gal和Fer-1对H9C2细胞内ROS的影响（×100）

Fig.1 The effect of D-gal and Fer-1 on ROS of H9C2 cells (×100)

表1 各组ROS、GSH、MDA含量比较（n=3，$\bar{x}±s$）

Tab.1 Comparison of ROS, GSH and MDA contents between the three groups

组别	ROS	GSH（μg/10⁶ cell）	MDA（nmol/mg prot）
Control组	15.24±1.65	86.01±5.73	0.58±0.05
D-gal组	39.90±4.83^a	63.06±10.98^a	1.46±0.15^a
Fer-1组	26.50±2.66^b	82.59±3.28^b	0.94±0.05^b
F	41.345^＊＊	8.408^＊	67.163^＊＊

图2 D-gal和Fer-1对H9C2细胞内P53、SLC7A11、GPx4蛋白表达的影响

Fig.2 The effects of D-gal and Fer-1 on protein expression levels of SLC7A11, GPx4 and P53 in H9C2 cells

表2 各组的P53、SLC7A11、GPx4蛋白表达水平比较

Tab.2 Comparison of protein expression levels of P53, SLC7A11 and GPx4 between the three groups

组别	SLC7A11	GPx4	P53
Control组	1.04±0.18	1.03±0.18	0.69±0.35
D-gal组	0.48±0.09^a	0.67±0.12^a	1.13±0.05^a
Fer-1组	0.79±0.12^b	0.98±0.07^b	0.88±0.04^b
F	12.706^＊＊	6.730^＊	92.479^＊＊

参考文献 29

[1]	中国心血管健康与疾病报告编写组. 中国心血管健康与疾病报告2020概要[J]. 中国循环杂志, 2021, 36(6):521-545.
	The Writing Committee of the Report on Cardiovascular Health and Diseases in China. Report on cardiovascular health and diseases burden in China:An updated su-mmary of 2020[J]. Chinese Circulation Journal, 2021, 36(6):521-545. doi:10.3969/j.issn.1000-3614.2021.06.001.
[2]	储睿, 朱刚艳, 周兴宇. 衰老与心血管疾病关系的研究进展[J]. 医学研究杂志, 2018, 47(7):187-190,58.
	CHU R, ZHU G Y, ZHOU X Y. Research progress on the relationship between aging and cardiovascular disease[J]. Journal of Medical Research, 2018, 47(7):187-190,58. doi:10.11969/j.issn.1673-548X.2018.07.044.
[3]	王曼怡, 朱家明, 孔昊. 基于Logistic模型对心血管疾病风险的预测[J]. 齐齐哈尔大学学报(自然科学版), 2017, 33(5):64-68.
	WANG M Y, ZHU J M, KONG H. Prediction of cardiovascular disease risk based on Logistic model[J]. Journal of Qiqihar University(Natural Science Edition),2017, 33(5):64-68. doi:10.3969/j.issn.1007-984X.2017.05.015.
[4]	LIU X L, ZHAO Y C, ZHU H Y, et al. Taxifolin retards the D-galactose-induced aging process through inhibiting Nrf2-mediated oxidative stress and regulating the gut microbiota in mice[J]. Food Funct, 2021, 12(23):12142-12158. doi:10.1039/d1fo01349a.
[5]	于春艳, 于春荣, 敬舒, 等. 北五味子总木脂素对D-半乳糖诱导的小鼠脑衰老自噬和凋亡的影响[J]. 吉林大学学报(医学版), 2014, 40(6):1210-1215.
	YU C Y, YU C R, JING S, et al. Effects of Schisandra total lignin on autophagy and apoptosis of mouse brain aging induced by D-galactose[J]. Journal of Jilin University (Medicine Edition), 2014, 40(6):1210-1215. doi:10.13481/j.1671-587x.20140618.
[6]	WANG D, WANG T, LI Z, et al. Green tea polyphenols upregulate the Nrf2 signaling pathway and suppress oxidative stress and inflammation markers in D-galactose-induced liver aging in mice[J]. Front Nutr, 2022, 9:836112. doi:10.3389/fnut.2022.836112.
[7]	CHEN X, LI D, SUN H Y, et al. Relieving ferroptosis may partially reverse neurodegeneration of the auditory cortex[J]. FEBS J, 2020, 287(21):4747-4766. doi:10.1111/febs.15266.
[8]	SUN C, PENG F, LI J, et al. Ferroptosis-specific inhibitor ferrostatin-1 relieves H₂O₂^-induced redox imbalance in primary cardiomyocytes through the Nrf2/ARE pathway[J]. Dis Markers, 2022, 2022:4539932. doi:10.1155/2022/4539932.
[9]	HUANG F, YANG R, XIAO Z, et al. Targeting ferroptosis to treat cardiovascular diseases: A new continent to be explored[J]. Front Cell Dev Biol, 2021, 9:737971. doi:10.3389/fcell.2021.737971.
[10]	刘建亚, 冯文静, 牟婕, 等. hUCMSCs对D-半乳糖致衰老小鼠心脏保护作用[J]. 青岛大学学报(医学版), 2018, 54(2):189-192,196.
	LIU J Y, FENG W J, MOU J, et al. Cardioprotective effect of human umbilical cord mesenchymal stem cells in D-galactose-induced aging mice[J]. Acta Aacademiae Medicinae Qingdao Universitatis, 2018, 54(2):189-192,196. doi:10.11712/jms201802015.
[11]	MAO H, ZHAO Y, LI H, et al. Ferroptosis as an emerging target in inflammatory diseases[J]. Prog Biophys Mol Biol, 2020, 155:20-28. doi:10.1016/j.pbiomolbio.2020.04.001.
[12]	TATARKOVÁ Z, KUKA S, RAČAY P, et al. Effects of aging on activities of mitochondrial electron transport chain complexes and oxidative damage in rat heart[J]. Physiol Res, 2011, 60(2):281-289. doi:10.33549/physiolres.932019.
[13]	TERMAN A, KURZ T, NAVRATIL M, et al. Mitochondrial turnover and aging of long-lived postmitotic cells:The mitochondrial-lysosomal axis theory of aging[J]. Antioxid Redox Signal, 2010, 12(4):503-535. doi:10.1089/ars.2009.2598.
[14]	LIANG W J, GUSTAFSSON Å B. The aging heart:Mitophagy at the center of rejuvenation[J]. Front Cardiovasc Med, 2020, 7:18. doi:10.3389/fcvm.2020.00018.
[15]	SHAKERI H, LEMMENS K, GEVAERT A B, et al. Cellular senescence links aging and diabetes in cardiovascular disease[J]. Am J Physiol Heart Circ Physiol, 2018, 315(3):H448-H462. doi:10.1152/ajpheart.00287.2018.
[16]	IZZO C, CARRIZZO A, ALFANO A, et al. The impact of aging on cardio and cerebrovascular diseases[J]. Int J Mol Sci, 2018, 19(2):481. doi:10.3390/ijms19020481.
[17]	SHIMIZU I, MINAMINO T. Cellular senescence in cardiac diseases[J]. J Cardiol, 2019, 74(4):313-319. doi:10.1016/j.jjcc.2019.05.002.
[18]	PIGNATELLI P, MENICHELLI D, PASTORI D, et al. Oxidative stress and cardiovascular disease:New insights[J]. Kardiol Pol, 2018, 76(4):713-722. doi:10.5603/KP.a2018.0071.
[19]	FENG W, LIU J, WANG S, et al. Alginate oligosaccharide alleviates D-galactose-induced cardiac ageing via regulating myocardial mitochondria function and integrity in mice[J]. J Cell Mol Med, 2021, 25(15):7157-7168. doi:10.1111/jcmm.16746.
[20]	SAHIN E, COLLA S, LIESA M, et al. Telomere dysfunction induces metabolic and mitochondrial compromise[J]. Nature, 2011, 470(7334):359-365. doi:10.1038/nature09787.
[21]	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. doi:10.1016/j.cell.2012.03.042.
[22]	STOCKWELL B R, FRIEDMANN ANGELI J P, BAYIR H, et al. Ferroptosis:A regulated cell death nexus linking metabolism,redox biology,and disease[J]. Cell, 2017, 171(2):273-285. doi:10.1016/j.cell.2017.09.021.
[23]	SUN Y, ZHENG Y, WANG C, et al. Glutathione depletion induces ferroptosis,autophagy, and premature cell senescence in retinal pigment epithelial cells[J]. Cell Death Dis, 2018, 9(7):753. doi:10.1038/s41419-018-0794-4.
[24]	FORCINA G C, DIXON S J. GPX4 at the crossroads of lipid homeostasis and ferroptosis[J]. Proteomics, 2019, 19(18):e1800311. doi:10.1002/pmic.201800311.
[25]	LIU P, FENG Y, LI H, et al. Ferrostatin-1 alleviates lipopolysaccharide-induced acute lung injury via inhibiting ferroptosis[J]. Cell Mol Biol Lett, 2020, 25:10. doi:10.1186/s11658-020-00205-0.
[26]	LARRICK J W, LARRICK J W, MENDELSOHN A R. Contribution of ferroptosis to aging and frailty[J]. Rejuvenation Res, 2020, 23(5):434-438. doi:10.1089/rej.2020.2390.
[27]	JENKINS N L, JAMES S A, SALIM A, et al. Changes in ferrous iron and glutathione promote ferroptosis and frailty in aging Caenorhabditis elegans[J]. Elife, 2020, 9:e56580. doi:10.7554/eLife.56580.
[28]	WU X, LI Y, ZHANG S, et al. Ferroptosis as a novel therapeutic target for cardiovascular disease[J]. Theranostics, 2021, 11(7):3052-3059. doi:10.7150/thno.54113.
[29]	RITZENTHALER J D, TORRES-GONZALEZ E, ZHENG Y, et al. The profibrotic and senescence phenotype of old lung fibroblasts is reversed or ameliorated by genetic and pharmacological manipulation of Slc7a11 expression[J]. Am J Physiol Lung Cell Mol Physiol, 2022, 322(3):L449-L461. doi:10.1152/ajplung.00593.2020.

Ferrostatin-1通过抑制铁死亡延缓D-gal诱导的心肌细胞衰老的研究

Ferrostatin-1 delay D-gal induced cardiomyocyte senescence by inhibiting ferroptosis

引用本文

RichHTML

PDF (PC)

可视化

摘要/Abstract

使用本文

图/表 4

参考文献 29

相关文章 15

编辑推荐

Metrics

本文评价

[1]	张春虹, 黄洪超, 刘越, 杜立龙, 许海委, 黎宁, 李勇进. 基于RNA测序和生物信息学分析鉴定椎旁肌退变中关键的铁死亡基因[J]. 天津医药, 2024, 52(9): 991-995.
[2]	刘斌, 杨龙, 李文莉, 邵宁宁, 董津睿. 小胶质细胞铁死亡在烟雾吸入性脑损伤中的作用机制探讨[J]. 天津医药, 2024, 52(8): 791-797.
[3]	王欣爽, 安亚娟, 管秀菊, 李娇, 刘玥, 魏丽萍, 齐新. 异甘草酸镁改善顺铂诱导的大鼠心肌损伤[J]. 天津医药, 2024, 52(8): 809-814.
[4]	黄晓蕾, 葛婷婷, 赵俊松, 倪志华. 人参皂苷Rg1在IL-6诱导的大鼠神经元铁死亡中的作用[J]. 天津医药, 2024, 52(11): 1137-1140.
[5]	李艳萍, 王协涛, 史立彬, 刘琼. 白藜芦醇调控Nrf2-GPX4通路对H₂O₂诱导肺泡上皮细胞铁死亡的影响[J]. 天津医药, 2023, 51(6): 568-572.
[6]	盛思琪, 谢琳, 姜怡邓, 熊建团, 杨安宁, 吴凯, 杨勇, 杨晓明. 转录因子EB在衰老心肌细胞自噬中的作用机制研究[J]. 天津医药, 2023, 51(3): 246-251.
[7]	刘艳秋, 范海迪, 侯海燕, 孙健, 林宁. sTIM-3及其配体Gal-9、HMGB1与2型糖尿病并发冠心病的相关性研究[J]. 天津医药, 2023, 51(3): 294-298.
[8]	胡英山, 王晶晶, 高红梅. 脓毒症相关肠道功能障碍发病机制的研究进展[J]. 天津医药, 2023, 51(3): 333-336.
[9]	姜花, 沈延梅, 马驯凯. 下调miR-106a-5p对过氧化氢诱导的心肌细胞损伤及JAK1/STAT3通路的影响[J]. 天津医药, 2023, 51(2): 113-117.
[10]	毛权西, 李作孝. 依达拉奉右莰醇通过铁死亡-脂质过氧化通路对脑出血大鼠神经保护的作用机制[J]. 天津医药, 2023, 51(11): 1199-1204.
[11]	王柯, 叶寒露. 大黄素对膝骨关节炎大鼠软骨细胞铁死亡的影响及机制研究[J]. 天津医药, 2023, 51(10): 1090-1097.
[12]	诸静, 冒丽静. 子宫内膜异位症患者血清中Gal-1和CRH水平变化及临床意义[J]. 天津医药, 2022, 50(9): 965-969.
[13]	郑君毅, 张莹莹, 刘园园, 陈孟英, 郭绪昆. FUNDC1通过调控线粒体分裂影响高糖损伤H9c2心肌细胞凋亡的机制研究[J]. 天津医药, 2022, 50(8): 791-795.
[14]	李秋畅, 闫顺昌, 蒙亚珍, 顾云霞, 宁巧明, 李娜, 王志华. Nrf2-GPX4介导的铁死亡通路参与右美托咪定对脑出血大鼠神经保护作用的机制研究[J]. 天津医药, 2022, 50(8): 817-821.
[15]	谭小兵, 王儒仙, 吕美荣, 孙显凤, 吕娜英, 戴青原. 人牙源性iPSCs分化心肌细胞促进急性心肌梗死小鼠心肌修复的研究[J]. 天津医药, 2022, 50(8): 822-826.