LAPTM5通过抑制铁死亡改善阿霉素诱导的心肌细胞损伤

张亚丹; 肖杨; 姚琪; 余嘉彬; 唐其柱

doi:10.14188/j.1671-8852.2024.0219

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LAPTM5通过抑制铁死亡改善阿霉素诱导的心肌细胞损伤

基础医学研究 | 更新时间：2026-11-05

- LAPTM5通过抑制铁死亡改善阿霉素诱导的心肌细胞损伤
- LAPTM5 protects against doxorubicin⁃induced cardiotoxicity by inhibiting ferroptosis
- 武汉大学学报（医学版） 2025年46卷第7期页码：829-835
- 作者机构：
  
  1.武汉大学人民医院心血管内科湖北武汉 430060
  2.代谢与相关慢病湖北省重点实验室湖北武汉 430060
- 作者简介：
  
  张亚丹，女，1999—，医学硕士生，主要从事心血管疾病的研究，E⁃mail：zhangyadan@whu.edu.cn
  唐其柱，男，1963—，医学博士，教授，主任医师，博士生导师，主要从事心血管及慢性疾病的研究，E⁃mail：qztang@whu.edu.cn
- 基金信息：
  
  国家自然科学基金区域创新发展联合基金项目(U22A20269)
- DOI：10.14188/j.1671-8852.2024.0219
  中图分类号： R54
- 收稿：2024-03-28，
  
  纸质出版：2025-07-25
- 稿件说明：
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张亚丹,肖杨,姚琪等.LAPTM5通过抑制铁死亡改善阿霉素诱导的心肌细胞损伤[J].武汉大学学报（医学版）,2025,46(07):829-835.

ZHANG Yadan,XIAO Yang,YAO Qi,et al.LAPTM5 protects against doxorubicin⁃induced cardiotoxicity by inhibiting ferroptosis[J].Medical Journal of Wuhan University,2025,46(07):829-835.
张亚丹,肖杨,姚琪等.LAPTM5通过抑制铁死亡改善阿霉素诱导的心肌细胞损伤[J].武汉大学学报（医学版）,2025,46(07):829-835. DOI： 10.14188/j.1671-8852.2024.0219.

ZHANG Yadan,XIAO Yang,YAO Qi,et al.LAPTM5 protects against doxorubicin⁃induced cardiotoxicity by inhibiting ferroptosis[J].Medical Journal of Wuhan University,2025,46(07):829-835. DOI： 10.14188/j.1671-8852.2024.0219.

摘要

目的

研究溶酶体相关跨膜蛋白5（LAPTM5）对阿霉素（DOX）诱导的心脏毒性的作用及机制。

方法

体外构建DOX损伤模型，将H9C2心肌细胞随机分为SiRNA⁃NC组

Si⁃LAPTM5组，SiRNA⁃NC+DOX组，Si⁃LAPTM5+DOX组4组。使用LiPo6000

转染试剂将SiRNA⁃NC(50 nmol•L

-1

)、Si⁃LAPTM5(50 nmol•L

-1

)病毒分别转染至H9C2心肌细胞，转染4 h后，使用1 μmol•L

-1

OX或等体积PBS刺激24 h构建心肌细胞损伤模型。采用Western Blot检测LAPTM5蛋白表达量，确认LAPTM5成功敲低，然后使用LDH及CK⁃MB试剂盒检测心肌细胞损伤情况，使用CCK⁃8试剂盒检测细胞存活率，TUNEL法检测心肌细胞凋亡水平；用DCFH⁃DA探针法和MDA、NADPH、SOD试剂盒检测氧化应激水平。采用Western Blot检测SLC7A11

、

GPX4

、

Ferritin等铁死亡相关蛋白的表达水平。

结果

Si⁃LAPTM5转染后，细胞中LAPTM5表达量下降约75%（

0.05）。DOX刺激可诱导心肌细胞损伤，使细胞LDH及CK⁃MB水平升高、存活率降低（

0.05），细胞凋亡增加。抑制LAPTM5表达可进一步加重DOX诱导的心肌细胞损伤（

0.05）。LAPTM5主要通过调节铁死亡发挥上述作用，DOX刺激明显上调细胞内Fe

含量和细胞膜上转铁蛋白TFR1水平，抑制LAPTM5会加剧细胞内Fe

蓄积，并进一步上调TFR1水平。DOX处理明显增加细胞氧化应激标志物的水平（ROS、脂质过氧化、MDA、NADPH酶活力）并降低SOD活性（

0.05），而抑制LAPTM5表达进一步恶化了DOX诱导的氧化应激（

0.05）。DOX刺激后铁死亡相关蛋白SLC7A11、GPX4表达水平下降，Ferritin表达水平升高（

0.05），抑制LAPTM5表达导致SLC7A11、GPX4蛋白表达水平进一步下降，Ferritin蛋白水平进一步升高。

结论

LAPTM5通过抑制铁死亡改善DOX诱导的心肌细胞毒性。

Abstract

Objective

To investigate the effects and mechanism of lysosomal⁃associated protein transmembrane 5 (LAPTM5) in doxorubicin (DOX)⁃induced cardiotoxicity.

Methods

H9C2 cardiomyocytes were cultured

in vitro

and randomly divided into the SiRNA⁃NC group

Si⁃LAPTM5 group

SiRNA⁃NC+DOX group

and Si⁃LAPTM5+DOX group. SiRNA⁃NC (50 nmol•L

-1

) and Si⁃LAPTM5 (50 nmol•L

-1

) were transfected into H9C2 cardiomyocytes using LiPo6000

transfection reagent for 4 h and then stimulated with 1 μmol•L

-1

DOX or the same volume of PBS for 24 h. The successful knockdown of LAPTM5 was confirmed by Western Blot. Then

myocardial cell injury was detected by lactate dehydrogenase (LDH) and creatine kinase isoenzyme MB (CK⁃MB) assay kit. Cell viability was detected by the cell counting kit⁃8 (CCK⁃8) assay kit. The Fe

levels were measured with a FerroOrange assay kit. The level of TFR1 was detected by immunofluorescence. The level of oxidativ

e stress was evaluated by 2′

7′dichlorodihydrofluorescein diacetate (DCFH⁃DA)

malondialdehyde (MDA) assay kit

NADPH oxidase assay kit

and SOD assay kit. The protein expression levels of SLC7A11

GPX4

and Ferritin were detected by Western Blot.

Results

After transfection with Si⁃LAPTM5

the expression of LAPTM5 in H9C2 cells decreased by about 75% (

0.05). LAPTM5 knockdown in H9C2 cells deteriorated DOX⁃induced cell injury and cell apoptosis

which was represented by further increased LDH level and CK⁃MB level

decreased cell viability

and increased TUNEL⁃positive cells (

0.05). Compared with the SiRNA⁃NC group

DOX increased the level of TFR1 and Fe

. Inhibition of LAPTM5 further increased the level of TFR1 and Fe

. Compared with the SiRNA⁃NC group

DOX increased reactive oxygen species (ROS) level

lipid peroxidation level

MDA content

and NADPH oxidase activity in H9C2 cardiomyocytes (

0.05)

which were aggravated by LAPTM5 knockdown(

0.05). Compared with the SiRNA⁃NC group

DOX decreased SOD activity

which was aggravated by LAPTM5 knockdown (

0.05). Compared with the SiRNA⁃NC group

DOX decreased the expression levels of SLC7A11 and glutathione peroxidase 4 (GPX4) and increased the level of ferritin protein (

0.05). Inhibition of LAPTM5 expression further decreased the expression levels of SLC7A11 and GPX4 and increased the level of ferritin protein.

Conclusion

LAPTM5 has a protective effect on doxorubicin⁃induced cardiotoxicity

and the mechanism may be related to the inhibition of ferroptosis.

关键词

Keywords

references

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