MiR-22-3p regulates hypoxia-induced mitochondrial homeostasis and apoptosis of pulmonary artery smooth muscle cells by targeting apoptosis-inducing factor，mitochondrion-associated 1

Xiang Yujing; Tang Huting; An Yong

doi:10.13406/j.cnki.cyxb.003884

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MiR-22-3p regulates hypoxia-induced mitochondrial homeostasis and apoptosis of pulmonary artery smooth muscle cells by targeting apoptosis-inducing factor，mitochondrion-associated 1

Basic research | 更新时间：2025-11-20

- MiR-22-3p regulates hypoxia-induced mitochondrial homeostasis and apoptosis of pulmonary artery smooth muscle cells by targeting apoptosis-inducing factor，mitochondrion-associated 1
- Journal of Chongqing Medical University Vol. 50, Issue 11, Pages: 1531-1540(2025)
- 作者机构：
  
  重庆医科大学附属儿童医院胸心外科/国家儿童健康与疾病临床医学研究中心/儿童发育疾病研究教育部重点实验室/ 结构性出生缺陷与器官修复重建重庆市重点实验室，重庆 400010
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.003884
  CLC： R473.72
- Received：07 August 2024，
  
  Revised：2024-12-06，
  
  Accepted：12 December 2024，
  
  Published：28 November 2025
- 稿件说明：
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向渝静,唐胡婷,安永.miR-22-3p通过靶向AIFM1调控低氧诱导的肺动脉平滑肌细胞线粒体稳态和凋亡[J].重庆医科大学学报,2025,50(11):1531-1540.

Xiang Yujing,Tang Huting,An Yong.MiR-22-3p regulates hypoxia-induced mitochondrial homeostasis and apoptosis of pulmonary artery smooth muscle cells by targeting apoptosis-inducing factor，mitochondrion-associated 1[J].Journal of Chongqing Medical University,2025,50(11):1531-1540.
向渝静,唐胡婷,安永.miR-22-3p通过靶向AIFM1调控低氧诱导的肺动脉平滑肌细胞线粒体稳态和凋亡[J].重庆医科大学学报,2025,50(11):1531-1540. DOI： 10.13406/j.cnki.cyxb.003884.

Xiang Yujing,Tang Huting,An Yong.MiR-22-3p regulates hypoxia-induced mitochondrial homeostasis and apoptosis of pulmonary artery smooth muscle cells by targeting apoptosis-inducing factor，mitochondrion-associated 1[J].Journal of Chongqing Medical University,2025,50(11):1531-1540. DOI： 10.13406/j.cnki.cyxb.003884.

摘要

目的

通过建立体外肺动脉高压（pulmonary arterial hypertension，PAH）模型，探讨miR-22-3p与线粒体凋亡诱导因子1（apoptosis-inducing factor，mitochondrion-associated 1，AIFM1）对低氧诱导下肺动脉平滑肌细胞（pulmonary artery smooth muscle cells，PASMCs）的线粒体稳态和凋亡的影响。

方法

通过低氧培养构建体外的PAH 模型，并调控AIFM1和miR-22-3p的表达使其升高或降低。实时荧光定量聚合酶链反应（reverse transcription quantitative polymerase chain reaction，RT-qPCR）和免疫印迹技术检测AIFM1、miR-22-3p和活化半胱氨酸蛋白酶-3（cleaved cysteine-aspartic protease-3，Cleaved Caspase-3）的表达水平。细胞计数试剂盒-8（cell counting kit-8，CCK-8）检测细胞增殖活性。流式细胞术检测细胞凋亡情况。通过线粒体超氧化物指示剂（mitochondrial superoxide indicator，mitoSOX）、腺嘌呤核苷三磷酸（adenosine triphosphate，ATP）试剂盒检测线粒体功能。线粒体动力学使用线粒体红色荧光探针（mito-tracker red cmxros，MitoTracker）测量线粒体周长变化。并且通过双荧光素酶报告基因实验分析验证AIFM1和miR-22-3p之间的相互作用。

结果

低氧诱导下，PASMCs的线粒体活性氧（reactive oxygen species，ROS）含量升高，ATP水平下降，线粒体裂变增加，细胞凋亡减少。AIFM1过表达能够改善线粒体稳态，并增加细胞凋亡。miR-22-3p负调控AIFM1，并逆转其过表达效应。

结论

miR-22-3p通过靶向AIFM1改善低氧诱导的PASMCs线粒体稳态、增殖活性和凋亡，旨在为肺动脉高压的预防和治疗提供潜在理论依据。

Abstract

Objective

To investigate the effect of miR-22-3p and apoptosis-inducing factor，mitochondrion-associated 1（AIFM1） on mitochondrial homeostasis and apoptosis of pulmonary artery smooth muscle cells（PASMCs） under hypoxic conditions by establishing an

in vitro

model of pulmonary hypertension（PAH）.

Methods

PASMCs were cultured under hypoxic conditions to establish an

in vitro

model of PAH，and the expression levels of AIFM1 and miR-22-3p were upregulated or downregulated. Reverse transcription quantitative polymerase chain reaction（RT-qPCR） and Western blotting were used to measure the expression levels of AIFM1，miR-22-3p，and cleaved cysteine-aspartic protease-3（Cleaved Caspase-3）. cell counting kit-8（CCK-8） assay was used to measure the proliferative activity of cells，and flow cytometry was used to measure cell apoptosis. Mitochondrial superoxide indicator（mitoSOX） and adenosine triphosphate（ATP） assay kits were used to observe mitochondrial function and dynamics，and Mito-Tracker Red CMXRos（MitoTracker） was used to measure the change in mitochondrial circumference. Dual-luciferase reporter assay was used to validate the interaction between AIFM1 and miR-22-3p.

Results

Hypoxia increased the content of mitochondrial ROS，reduced the level of ATP，promoted mitochondrial fission，and reduced cell apoptosis in PASMCs. AIFM1 overexpression improved mitochondrial homeostasis and increased cell apoptosis，while miR-22-3p negatively regulated AIFM1 and reversed the effect of AIFM1 overexpression.

Conclusion

This study shows that miR-22-3p enhances mitochondrial homeostasis，proliferation，and apoptosis in hypoxia-induced PASMCs by targeting AIFM1，which provides a potential theoretical basis for the prevention and treatment of PAH.

关键词

Keywords

references

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