NCOA4介导的铁蛋白自噬参与LPS诱导的心肌细胞铁死亡

郭立春; 曹国栋; 罗茜晴; 林靓; 赵宇含; 曾佑成; 张艺馨; 程青虹

doi:10.16016/j.2097-0927.202204140

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NCOA4介导的铁蛋白自噬参与LPS诱导的心肌细胞铁死亡

基础医学 | 更新时间：2026-01-09

- NCOA4介导的铁蛋白自噬参与LPS诱导的心肌细胞铁死亡
- Nuclear receptor coactivator 4 -mediated ferritinophagy is involved in LPS-induced ferroptosis in cardiomyocytes
- 陆军军医大学学报 2022年44卷第23期页码：2401-2408
- 作者机构：
  
  1.832000 新疆石河子，石河子大学医学院
  2.832000 新疆石河子，石河子大学医学院第一附属医院重症二科
- 作者简介：
  
  [程青虹，E-mail: xunfeicheng2015@163.com CHENG Qinghong, E-mail: xunfeicheng2015@163.com
- 基金信息：
  
  国家自然科学基金地区科学基金项目(81860336);兵团科技攻关与成果转化计划项目(2016AD003)
- DOI：10.16016/j.2097-0927.202204140
  中图分类号： R322.11;R361.3;R631.2
- 收稿：2022-04-29，
  
  修回：2022-06-30，
  
  纸质出版：2022-12-15
- 稿件说明：
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郭立春, 曹国栋, 罗茜晴, 等. NCOA4介导的铁蛋白自噬参与LPS诱导的心肌细胞铁死亡[J]. 陆军军医大学学报, 2022,44(23):2401-2408.

Lichun GUO, Guodong CAO, Xiqing LUO, et al. Nuclear receptor coactivator 4 -mediated ferritinophagy is involved in LPS-induced ferroptosis in cardiomyocytes[J]. Journal of Army Medical University, 2022, 44(23): 2401-2408.
郭立春, 曹国栋, 罗茜晴, 等. NCOA4介导的铁蛋白自噬参与LPS诱导的心肌细胞铁死亡[J]. 陆军军医大学学报, 2022,44(23):2401-2408. DOI： 10.16016/j.2097-0927.202204140.

Lichun GUO, Guodong CAO, Xiqing LUO, et al. Nuclear receptor coactivator 4 -mediated ferritinophagy is involved in LPS-induced ferroptosis in cardiomyocytes[J]. Journal of Army Medical University, 2022, 44(23): 2401-2408. DOI： 10.16016/j.2097-0927.202204140.

摘要

目的

探讨核受体辅激活因子4(nuclear receptor coactivator 4，NCOA4)介导的铁蛋白(Ferritin)自噬是否参与脂多糖(lipopolysaccharide，LPS)诱导的脓毒症心肌细胞铁死亡的发生。

方法

采用不同剂量(5、10、20 μg/mL)的LPS诱导大鼠心肌细胞H9c2形成脓毒症心肌细胞损伤体外模型，以正常H9c2细胞作为Control组。应用试剂盒检测心肌细胞活力、Fe

、心肌酶释放水平变化与氧化应激水平；荧光探针检测细胞线粒体膜电位水平变化；免疫荧光检测NCOA4、Ferritin在心肌细胞中的表达定位；应用小干扰RNA(si RNA)降低H9c2细胞中的NCOA4的表达，Western blot检测铁死亡调控蛋白、自噬蛋白表达水平。

结果

不同剂量LPS刺激后H9c2细胞活力明显降低(

＜0.05)，Fe

浓度升高(

＜0.05)，丙二醛(Malondialdehyde，MDA)、活性氧(reactive oxygen species，ROS)等细胞氧化应激的水平升高(

＜0.05)，线粒体膜电位降低(

＜0.05)，荧光镜下观察到NCOA4-Ferritin共定位表达增加(

＜0.05)，铁死亡调控蛋白NCOA4表达增加(

＜0.05)，Ferritin、SLC7A11、GPX4的表达水平下降(

＜0.05)，自噬蛋白LC3-Ⅱ表达增加(

＜0.05)。慢病毒敲低NCOA4表达，Ferritin、SLC7A11、GPX4的表达增加(

＜0.05)，LC3-Ⅱ表达水平下降(

＜0.05)。

结论

NCOA4介导的铁蛋白自噬参与了脓毒症心肌细胞铁死亡的发生过程。

Abstract

Objective

To investigate whether ferritin autophagy mediated by nuclear receptor coactivator 4 (NCOA4) is involved in lipopolysaccharide (LPS)-induced ferroptosis in septic cardiomyocytes.

Methods

Different doses of LPS (5

10 and 20 μg/mL) were adopted to induce rat cardiomyocytes H9c2 to form an in vitro model of cardiomyocyte injury

and normal H9c2 cells were used as control group. The cell viability

level

releases of myocardial enzymes and oxidative stress level were detected by corresponding reagent kits

and the change in mitochondrial membrane potential was measured with fluorescent probe. Immunofluorescence assay was performed to determine the expression and localization of NCOA4 and ferritin in the cardiomyocytes. Moreover

small interfering RNA (siRNA) was used to reduce NCOA4 expression in H9c2 cells

and the levels of ferroptosis regulatory proteins as well as autophagy proteins were detected with Western blotting.

Results

After stimulation with different doses of LPS

the viability of H9c2 cells was decreased significantly (

0.05)

the concentration of Fe

was increased (

0.05)

the levels of oxidative stress products such as malondialdehyde (MDA) and reactive oxygen species (ROS) were both elevated (

0.05)

and the mitochondrial membrane potential was declined (

0.05). Increased expression of NCOA4-Ferritin colocalization was observed under fluoroscopy (

0.05). Western blotting showed that the expression levels of ferroptosis regulatory protein NCOA4 and autophagy protein LC3-Ⅱwere up-regulated (

0.05)

while those of ferritin

SLC7A11 and GPX4 were down-regulated (

0.05). NCOA4 knockdown resulted in significantly enhanced expression of ferritin

SLC7A11 and GPx4 (

0.05)

whereas reduced expression of LC3-Ⅱ (

0.05).

Conclusion

NCOA44-mediated ferritin autophagy is involved in the occurrence of ferroptosis in septic cardiomyocytes.

关键词

Keywords

references

EVANS L, RHODES A, ALHAZZANI W, et al . Surviving Sepsis campaign: international guidelines for management of Sepsis and septic shock 2021[J ] . Intensive Care Med, 2021, 47(11): 1181-1247. DOI: 10.1007/s00134-021-06506-y..

FLEISCHMANN-STRUZEK C, MELLHAMMAR L, ROSE N, et al . Incidence and mortality of hospital- and ICU-treated Sepsis : results from an updated and expanded systematic review and meta-analysis[J ] . Intensive Care Med, 2020, 46(8): 1552-1562. DOI: 10.1007/s00134-020-06151-x..

赵鹏, 寿松涛, 李晨, 等. 脓毒症患者合并心肌损伤的危险因素及其生存分析[J]. 中华医院感染学杂志, 2022, 32(7): 961-965.

ZHAO P, SHOU S T, LI C, et al . Risk factors for myocardial injury in Sepsis patients and survival analysis[J ] . Chin J Nosocomiology, 2022, 32(7): 961-965..

HOLLENBERG S M, SINGER M. Pathophysiology of Sepsis -induced cardiomyopathy[J ] . Nat Rev Cardiol, 2021, 18(6): 424-434. DOI: 10.1038/s41569-020-00492-2.

ZHENG J S, CONRAD M. The metabolic underpinnings of ferroptosis[J]. Cell Metab, 2020, 32(6): 920-937. DOI: 10.1016/j.cmet.2020.10.011.

SANTANA-CODINA N, GIKANDI A, MANCIAS J D. The role of NCOA4-mediated ferritinophagy in ferroptosis[J]. Adv Exp Med Biol, 2021, 1301: 41-57. DOI: 10.1007/978-3-030-62026-4_4.

QIU Y M, CAO Y, CAO W J, et al . The application of ferroptosis in diseases[J ] . Pharmacol Res, 2020, 159: 104919. DOI: 10.1016/j.phrs.2020.104919.

WU X G, LI Y, ZHANG S C, et al . Ferroptosis as a novel therapeutic target for cardiovascular disease[J ] . Theranostics, 2021, 11(7): 3052-3059. DOI: 10.7150/thno.54113.

LIU Y T, TAN S C, WU Y B, et al . The emerging role of ferroptosis in Sepsis [J ] . DNA Cell Biol, 2022, 41(4): 368-380. DOI: 10.1089/dna.2021.1072..

REN Q, ZHAO S J, REN C J, et al . Astragalus polysaccharide alleviates LPS-induced inflammation injury by regulating miR-127 in H9c2 cardiomyoblasts[J ] . Int J Immunopathol Pharmacol, 2018, 32: 2058738418759180. DOI: 10.1177/2058738418759180..

FONT M D, THYAGARAJAN B, KHANNA A K. Sepsis and Septic Shock-Basics of diagnosis, pathophysiology and clinical decision making[J ] . Med Clin North Am, 2020, 104(4): 573-585. DOI: 10.1016/j.mcna.2020.02.011.

慕婉晴, 韩奕, 顾国嵘, 等. 脓毒症患者的预后危险因素及脓毒症相关性心肌损伤患者的临床特征研究[J]. 中华危重病急救医学, 2021(7): 809-814.

MU W Q, HAN Y, GU G R, et al . Prognostic risk factors of patients with Sepsis and the clinical characteristics of patients with septic myocardial injury[J ] . Chin Crit Care Med, 2021(7): 809-814..

DICKSON K, LEHMANN C. Inflammatory response to different toxins in experimental Sepsis models[J ] . Int J Mol Sci, 2019, 20(18): E4341. DOI: 10.3390/ijms20184341.

LU J Q, LIU J Y, LI A. Roles of neutrophil reactive oxygen species (ROS) generation in organ function impairment in Sepsis [J ] . J Zhejiang Univ Sci B, 2022, 23(6): 437-450. DOI: 10.1631/jzus.B2101075.

LIU T, SUN L, ZHANG Y B, et al . Imbalanced GSH/ROS and sequential cell death[J ] . J Biochem Mol Toxicol, 2022, 36(1): e22942. DOI: 10.1002/jbt.22942.

SUN Y T, CHEN P, ZHAI B T, et al . The emerging role of ferroptosis in inflammation[J ] . Biomedecine Pharmacother, 2020, 127: 110108. DOI: 10.1016/j.biopha.2020.110108.

李浩甲, 杨文曲, 韩冲芳, 等. 去铁胺对糖尿病大鼠心肌细胞铁死亡通路的影响[J]. 医学研究杂志, 2020, 49(9): 116-119. DOI: 10.11969/j.issn.1673-548X.2020.09.025.

LI H J, YANG W Q, HAN C F, et al . Effect of deferoxamine on ferroptosis of myocardial cell in diabetic rats[J ] . J Med Res, 2020, 49(9): 116-119. DOI: 10.11969/j.issn.1673-548X.2020.09.025.

LI J H, LI M Y, LI L, et al . Hydrogen sulfide attenuates ferroptosis and stimulates autophagy by blocking mTOR signaling in Sepsis -induced acute lung injury[J ] . Mol Immunol, 2022, 141: 318-327. DOI: 10.1016/j.molimm.2021.12.003..

CHEN Y H, JIN S, TENG X, et al . Hydrogen sulfide attenuates LPS-induced acute kidney injury by inhibiting inflammation and oxidative stress[J ] . Oxid Med Cell Longev, 2018, 2018: 6717212. DOI: 10.1155/2018/6717212.

梁译丹, 覃王, 黄豪, 等. 自噬通过降解铁蛋白促进神经元铁死亡参与蛛网膜下腔出血后早期脑损伤[J]. 第三军医大学学报, 2019, 41(15): 1407-1414. DOI: 10.16016/j.1000-5404.201902124.

LIANG Y D, QIN W, HUANG H, et al . Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats[J ] . J Third Mil Med Univ, 2019, 41(15): 1407-1414. DOI: 10.16016/j.1000-5404.201902124.

HOU W, XIE Y C, SONG X X, et al . Autophagy promotes ferroptosis by deg radation of ferritin[J ] . Autophagy, 2016, 12(8): 1425-1428. DOI: 10.1080/15548627.2016.1187366.

ITO J, OMIYA S, RUSU M C, et al . Iron derived from autophagy-mediated ferritin degradation induces cardiomyocyte death and heart failure in mice[J ] . Elife, 2021, 10: e62174. DOI: 10.7554/eLife.62174.

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