MiR-32-3p improves ISO induced AC16 myocardial cell injury by targeting ATF4

Q: What are the key findings of this study?

Overexpression of miR-32-3p alleviated isoproterenol-induced AC16 cell damage, inflammation, and oxidative stress by directly targeting ATF4.

Q: What is the clinical evidence from this study?

Population: Myocardial cell injury and hypertrophy. Intervention: miR-32-3p mimic vs. Isoproterenol (ISO) + mimic NC. Primary outcome: Cell proliferation, hypertrophy markers, inflammatory cytokines, and oxidative stress (p=<0.0001).

Q: What does this research mean for the field?

Overexpression of miR-32-3p alleviates isoproterenol-induced cardiomyocyte damage, inflammation, and oxidative stress by directly targeting ATF4. Novelty: ClaimNovelty.INCREMENTAL. Consensus alignment: ConsensusAlignment.NEUTRAL.

Q: What question did this study set out to answer?

To investigate the expression pattern, functional role, and molecular mechanism of the miR-32-3p/ATF4 axis in isoproterenol-induced human AC16 cardiomyocyte injury.

Danqi Chen; Dongjun Hu; Xiongxiong Wang; Bangsheng Chen; Wenzhu Lou; Yuehua Chen

doi:10.1186/s13019-026-04395-0

July 2, 2026Journal of Cardiothoracic SurgeryOpen Access

MiR-32-3p improves ISO induced AC16 myocardial cell injury by targeting ATF4

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Key result

miR-32-3p overexpression alleviates isoproterenol-induced cardiomyocyte injury and inflammation by targeting ATF4.

P<0.0001

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Why the study?

Pathological myocardial hypertrophy is a major risk factor for heart failure, and the role of the miR-32-3p/ATF4 axis in ISO-induced cardiomyocyte injury remained to be clarified.

Does miR-32-3p overexpression alleviate isoproterenol-induced AC16 myocardial cell injury?

Key Points

To investigate the expression pattern, functional role, and molecular mechanism of the miR-32-3p/ATF4 axis in isoproterenol-induced human AC16 cardiomyocyte injury.
Cultured human AC16 cardiomyocytes were exposed to 1 µM isoproterenol (ISO) to establish an in vitro myocardial injury model.
Cells were transfected with miR-32-3p mimic, inhibitor, or ATF4 overexpression plasmids (oe-ATF4), followed by evaluation of cell proliferation (CCK-8), inflammatory cytokines (ELISA for IL-6, TNF-α, IL-1β), and oxidative stress markers (MDA, SOD, ROS).
Predicted interaction sites using TargetScanHuman and validated direct binding between miR-32-3p and the 3'UTR of ATF4 using dual-luciferase reporter gene assays.
ISO exposure significantly downregulated miR-32-3p (P < 0.0001), inhibited AC16 cell proliferation (P < 0.0001), upregulated hypertrophy markers ANP and β-MHC (P < 0.0001), and elevated levels of IL-6, TNF-α, IL-1β, MDA, and ROS alongside reduced SOD activity (all P < 0.0001).
Transfection with miR-32-3p mimic reversed ISO-induced damage, significantly downregulating ANP (P < 0.01), β-MHC (P < 0.0001), inflammatory cytokines (P < 0.0001), and MDA (P < 0.0001) while restoring cell viability (P < 0.0001) and SOD activity (P < 0.001).
Dual-luciferase assays confirmed miR-32-3p directly binds the 3'UTR of ATF4 (P < 0.0001), and co-transfection with oe-ATF4 eliminated the protective anti-hypertrophic, anti-inflammatory, and antioxidant effects of miR-32-3p (P < 0.05).

Structured PICO

Does miR-32-3p overexpression alleviate isoproterenol-induced AC16 myocardial cell injury?

P

Population

In vitro study using human AC16 cardiomyocytes to model isoproterenol-induced myocardial injury and hypertrophy.

I

Intervention

Transfection with miR-32-3p mimic or miR-32-3p inhibitor.

C

Comparator

Cells cultured in serum medium without ISO, and cells transfected with respective negative controls (miR-NC, inhibitor NC).

O

Outcome

Cell proliferation, expression of myocardial hypertrophy markers (ANP and β-MHC), levels of inflammatory cytokines (IL-6, TNF-α, IL-1β), and oxidative stress indicators (MDA, SOD, ROS).surrogate

Main Result

p-value: p=<0.0001

miR-32-3p protects against isoproterenol-induced cardiomyocyte injury, inflammation, and oxidative stress in vitro by directly targeting and negatively regulating ATF4.

Limitations

In vitro experiments were only conducted on AC16 cardiomyocytes, lacking validation of the in vivo function of the miR-32-3p/ATF4 axis in animal models.
Lack of further exploration of specific downstream effector pathways of ATF4.
Lack of clinical sample validation makes it difficult to clarify the expression characteristics of miR-32-3p in patients with myocardial hypertrophy and its correlation with disease severity and prognosis.
Lack of clinical sample validation makes it difficult to clarify the expression characteristics of miR-32-3p in patients with myocardial hypertrophy.

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Implication

No immediate clinical application; leaves open miR-32-3p/ATF4 targeting in cardiac injury pending in vivo validation.

Abstract

BACKGROUND: Pathological myocardial hypertrophy is a key risk factor for heart failure and cardiovascular death. MicroRNAs (miRNAs), as a key post transcriptional regulator of gene expression, play an increasingly important role in myocardial hypertrophy and injury. This study aimed to investigate the expression, function, and mechanism of miR-32-3p through the miR-32-3p/ATF4 axis in ISO induced injury of human AC16 cardiomyocytes. METHODS: A cell injury model was established by treating AC16 cells with 1 µM ISO. miR-32-3p mimic/inhibitor were transfected. Gene expression levels were detected using qRT-PCR; cell proliferation was evaluated via the CCK-8 assay; the levels of inflammatory cytokines (IL-6, TNF-α, IL-1β) were detected by ELISA; changes in oxidative stress levels (MDA, SOD, ROS) were analyzed using reagent kits; and the binding site between miR-32-3p and ATF4 was predicted through TargetScanHuman.; The miR-32-3p/ATF4 target relationship was validated using dual luciferase assays. RESULTS: ISO induced downregulation of miR-32-3p expression (P < 0.0001), inhibition of AC16 cell proliferation (P < 0.0001), upregulation of myocardial hypertrophy markers ANP and β-MHC, as well as increased levels of inflammatory cytokines IL-6, TNF-α, IL-1β and oxidative stress (all P < 0.0001). When the miR-32-3p mimic was added, these changes were reversed. The dual luciferase assay confirmed that bioinformatics predicted miR-32-3p to directly target the 3'UTR of ATF4. The rescue experiment showed that overexpression of ATF4 eliminated the protective effect of miR-32-3p (P < 0.05). CONCLUSIONS: miR-32-3p alleviates ISO induced AC16 cell damage by targeting ATF4, regulating inflammation and oxidative stress.

Discussion

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Cite This Study

Chen et al. (2026) studied Myocardial cell injury and hypertrophy. miR-32-3p mimic vs. Isoproterenol (ISO) + mimic NC was evaluated on Cell proliferation, hypertrophy markers, inflammatory cytokines, and oxidative stress (p=<0.0001). Overexpression of miR-32-3p alleviated isoproterenol-induced AC16 cell damage, inflammation, and oxidative stress by directly targeting ATF4.

synapsesocial.com/papers/6a49f411f5d1d45b287ffd32 https://doi.org/10.1186/s13019-026-04395-0

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MiR-32-3p improves ISO induced AC16 myocardial cell injury by targeting ATF4

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Key Points

Structured PICO

Main Result

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

MiR-32-3p improves ISO induced AC16 myocardial cell injury by targeting ATF4

Figures

Key Points

Structured PICO

Main Result

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

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