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MSCs-Derived Extracellular Vesicles Carrying miR-212-5p Alleviate Myocardial Infarction-Induced Cardiac Fibrosis via NLRC5/VEGF/TGF-β1/SMAD Axis

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Abstract

The purpose of the present study was to define the role of mesenchymal stem cell (MSC)–derived extracellular vesicles (EVs) in the progression of myocardial infarction (MI)–induced cardiac fibrosis. An in vitro cell model of hypoxia-induced cardiac fibrosis was constructed in cardiac fibroblasts (CFs). miR-212-5p was poorly expressed in clinical pathological samples and animal models of cardiac fibrosis caused by MI, while miR-212-5p expression was enriched in EVs released from MSCs. EVs from MSCs were isolated, evaluated, and co-cultured with CFs. Dual-luciferase reporter gene assay revealed that miR-212-5p negatively targeted NLRC5 progression of cardiac fibrosis. Following loss- and gain-function assay, EVs expressing miR-212-5p protected against cardiac fibrosis evidenced by reduced levels of α-SMA, Collagen I, TGF-β1, and IL-1β. In vivo experiments further confirmed the above research results. Collectively, EVs from MSCs expressing miR-212-5p may attenuate MI by suppressing the NLRC5/VEGF/TGF-β1/SMAD axis.

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Abbreviations

MSCs:

Mesenchymal stem cells

EVs:

Extracellular vesicles

MI:

Myocardial infarction

CFs:

Cardiac fibroblasts

MI:

Myocardial infarction

SPF:

Specific pathogen free

DMEM:

Dulbecco’s modified eagle medium

FITC:

Fluorescein isothiocyanate

PE:

Polyethylene

NC:

Negative control

BSA:

Bull serum albumin

ECG:

Echocardiogram

LVIDs:

Left ventricular inner systolic diameter

LVESV:

Left ventricular systolic volume

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Funding

This work was supported by the Medical Scientific Research Foundation of Guangdong Province (No. A2020002) and the Science and Technology Program of Guangzhou, China (No. 202102080011).

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Authors and Affiliations

  1. Department of Intensive Care Unit of Cardiac Surgery, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangdong Cardiovascular Institute, No. 96, Dongchuan Road, Guangzhou, 510080, People’s Republic of China

    Yijin Wu, Wenying Peng, Miaoxian Fang, Meifen Wu & Min Wu

  2. Department of Cardiac Surgery, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangdong Cardiovascular Institute, Guangzhou, 510080, People’s Republic of China

    Min Wu

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  1. Yijin Wu
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Correspondence to Min Wu.

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Associate Editor Junjie Xiao oversaw the review of this article

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Supplementary Information

Figure S1

Western blots. A, NLRC5, α-SMA and VEGF protein expression in cells tested by western blot analysis. B, The expression of p-SMAD2/3, NLRC5 and VEGF in anti-miR-212-5p and GW4869 groups detected by western blot analysis.) (PNG 5043 kb)

High resolution image (EPS 4752 kb)

Figure S2

MSCs-derived EVs deliver miR-212-5p to alleviate MI. A, TTC staining of the heart tissues. B, Statistics of TTC staining results. C, The serum cTnT level detected using ELISA (***p < 0.05 vs. the sham group; ###p < 0.05 vs. the MI group; &&&p < 0.05 vs. the MI-EVs-NC group). n = 8 in each group of mice. Measurement data were expressed by mean ± standard deviation. One-way ANOVA was used for multiple group comparisons, followed by Tukey's post hoc test. (PNG 2633 kb)

High resolution image (EPS 3924 kb)

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Wu, Y., Peng, W., Fang, M. et al. MSCs-Derived Extracellular Vesicles Carrying miR-212-5p Alleviate Myocardial Infarction-Induced Cardiac Fibrosis via NLRC5/VEGF/TGF-β1/SMAD Axis. J. of Cardiovasc. Trans. Res. 15, 302–316 (2022). https://doi.org/10.1007/s12265-021-10156-2

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