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Knockdown of Mmu-circ-0001380 Attenuates Myocardial Ischemia/Reperfusion Injury via Modulating miR-106b-5p/Phlpp2 Axis

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Abstract

Myocardial ischemia/reperfusion (MI/R) injury induces myocardial damage and dysfunction. Increasing evidence has confirmed that circular RNAs (circRNAs) play crucial roles in regulating MI/R. Mmu-circ-0001380 has identified to be highly expressed in myocardium of MI/R mouse model. However, its biological function and molecular mechanism in MI/R injury are still unclear. Here, we demonstrated that knockdown of cric-0001380 attenuated myocardial injury of MI/R mice. In vitro, silence of circ-0001380 significantly enhanced viability, and inhibited apoptosis and oxidative stress in HL-1 cells under oxygen–glucose deprivation/reoxygenation (OGD/R). Mmu-miR-106b-5p interacted with circ-0001380, and suppressed the expression of pleckstrin homology domain and leucine rich repeat protein phosphatase 2 (Phlpp2). The miR-106b-5p/Phlpp2 axis mediated the effect of circ-0001380 on OGD/R-induced apoptosis through regulating the phosphorylation of p38, and further involved in regulating the viability and oxidative stress of HL-1 cells. In conclusion, circ-0001380 downregulation relieves MI/R injury via regulating the miR-106b-5p/Phlpp2 axis.

Graphical Abstract

The present study indicates that mmu-circ-0001380 exacerbates the myocardial ischemia/reperfusion injury through modulating the miR-106b-5p/Phlpp2 axis in vitro and in vivo.

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Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

MI/R:

Myocardial ischemia/reperfusion

circRNA:

Circular RNA

OGD/R:

Oxygen-glucose deprivation/reoxygenation

AMI:

Acute myocardial infarction

ROS:

Reactive oxygen species

ceRNA:

Competitive endogenous RNAs

Phlpp2:

PH domain and leucine rich repeat protein phosphatase 2

MDA:

Malonaldehyde

SOD:

Superoxide dismutase

GSH-Px:

Glutathione peroxidase

CK-MB:

Creatine kinase myocardial band

LDH:

Lactate dehydrogenase

I/R:

Ischemia/reperfusion

LVEDD:

Left ventricular end-diastolic dimension

LVESD:

Left ventricular end systolic diameter

LVEF:

Left ventricular ejection fraction

LVFS:

Left ventricular fraction shortening

cDNA:

Complementary DNA

gDNA:

Genomic DNA

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Funding

This research was supported by Dalian Municipal Central Hospital Gaofeng Plan of Dalian Key Medical Specialty ‘Dengfeng Plan’ at Department of Cardiology.

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Authors and Affiliations

  1. Department of Cardiology, Dalian Municipal Central Hospital, No. 826, Xinan Road, Dalian, Liaoning, China

    Li Wang

  2. Department of Cardiology, Shengjing Hospital of China Medical University, No. 39, Huaxiang Road, Shenyang, Liaoning, China

    Chuanhe Wang, Zhaoqing Sun, Aolin Du, Fei Shan & Zhijun Sun

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Correspondence to Li Wang or Zhijun Sun.

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Fig. S1

Downregulation of miR-106b-5p inhibits cell viability and aggravates oxidative stress and apoptosis through regulating Phlpp2. A. Real-time PCR evaluated the expression of miR-106b-5p in HL-1 cells. B. The mRNA level of Phlpp2 was detected by real-time PCR. C. Phlpp2 protein expression was assessed using western blot. D. The knockdown efficiency of siPhlpp2 was examined using real-time PCR. E and F. The mRNA and protein expression of Phlpp2 was estimated in OGD/R-induced HL-1 cells. G. MDA content and SOD activity were evaluated using the corresponding kits. H. Cell viability was examined using MTT assay in each group. I. Cell apoptosis was detected by TUNEL assay. Scale bars = 50 μm. There were three biological replicates in each group. Data are presented as mean ± SD. * P < 0.05; ** P < 0.01; ns, P > 0.05. (PNG 562 kb)

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Wang, L., Wang, C., Sun, Z. et al. Knockdown of Mmu-circ-0001380 Attenuates Myocardial Ischemia/Reperfusion Injury via Modulating miR-106b-5p/Phlpp2 Axis. J. of Cardiovasc. Trans. Res. 16, 1064–1077 (2023). https://doi.org/10.1007/s12265-023-10383-9

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