Nuclear receptor coactivator 4 -mediated ferritinophagy is involved in LPS-induced ferroptosis in cardiomyocytes

Lichun GUO; Guodong CAO; Xiqing LUO; Jing LIN; Yuhan ZHAO; Youcheng ZENG; Yixin ZHANG; Qinghong CHENG

doi:10.16016/j.2097-0927.202204140

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Nuclear receptor coactivator 4 -mediated ferritinophagy is involved in LPS-induced ferroptosis in cardiomyocytes

Basic Medicine | 更新时间：2026-01-09

- Nuclear receptor coactivator 4 -mediated ferritinophagy is involved in LPS-induced ferroptosis in cardiomyocytes
- Journal of Army Medical University Vol. 44, Issue 23, Pages: 2401-2408(2022)
- 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：
- 基金信息：
  
  the Regional Science Fund Project of National Natural Science Foundation of China(81860336);the Project of Science and Technology Tackling and Achievement Transformation Plan of Xinjiang Corps(2016AD003)
- DOI：10.16016/j.2097-0927.202204140
  CLC： R322.11;R361.3;R631.2
- Received：29 April 2022，
  
  Revised：2022-06-30，
  
  Published：15 December 2022
- 稿件说明：
移动端阅览
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：

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

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