https://orcid.org/0000-0003-3147-9934
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Abstract
Background
Cardiac repair remains a thorny issue for survivors of acute myocardial infarction (AMI), due to the regenerative inertia of myocardial cells. Cell-free therapies, such as exosome transplantation, have become a potential strategy for myocardial injury. The aim of this study was to investigate the role of engineered exosomes in overexpressing Growth Differentiation Factor-15 (GDF-15) (GDF15-EVs) after myocardial injury, and their molecular mechanisms in cardiac repair.
Methods
H9C2 cells were transfected with GDF-15 lentivirus or negative control. The exosomes secreted from H9C2 cells were collected and identified. The cellular apoptosis and autophagy of H2O2-injured H9C2 cells were assessed by Western blotting, TUNEL assay, electron microscopy, CCK-8 and caspase 3/7 assay. A rat model of AMI was constructed by ligating the left anterior descending artery. The anti-apoptotic, pro-angiogenic effects of GDF15-EVs treatment, as well as ensuing functional and histological recovery were evaluated. Then, mRNA sequencing was performed to identify the differentially expressed mRNAs after GDF15-EVs treatment.
Results
GDF15-EVs inhibited apoptosis and promoted autophagy in H2O2 injured H9C2 cells. GDF15-EVs effectively decreased the infarct area and enhanced the cardiac function in rats with AMI. Moreover, GDF15-EVs hindered inflammatory cell infiltration, inhibited cell apoptosis, and promoted cardiac angiogenesis in rats with AMI. RNA sequence showed that telomerase reverse transcriptase (TERT) mRNA was upregulated in GDF15-EVs-treated H9C2 cells. AMPK signaling was activated after GDF15-EVs. Silencing TERT impaired the protective effects of GDF15-EVs on H2O2-injured H9C2 cells.
Conclusion
GDF15-EVs could fulfil their protective effects against myocardial injury by upregulating the expression of TERT and activating the AMPK signaling pathway. GDF15-EVs might be exploited to design new therapies for AMI.
Abbreviations
AMI, acute myocardial infarction; GDF15-EVs, extracellular vesicles of H9C2 cells with stable overexpression of GDF-15; NC-EVs: extracellular vesicles of control H9C2 cells; MI, myocardial infarction; HF, heart failure; PCI, Percutaneous transluminal coronary intervention; mRNA, messenger ribonucleic acid; FBS, fetal bovine serum; NTA, nanoparticle tracking analysis; TEM, transmission electron microscopy; PBS, phosphate buffered saline; SDS, sodium dodecyl sulfate; CCK-8, Cell Counting Kit-8; PI, propidium iodide; RT-qPCR, quantitative real-time PCR; TSG101, tumor susceptibility gene 101 protein; CD63, lysosome-associated membrane protein 3; CD81, target of an antiproliferative antibody-1; BAX, BCL2L4; Bcl-2, B-cell lymphoma-2; P62, Sequestosome 1; Lc3, Map1LC3; TNF-α, tumor necrosis factor-α; IL-10, interleukin-10; IL-6, interleukin-6; β-actin, beta-actin; β-actin, beta-actin; GAPDH, Glyceraldehyde-3-phosphate dehydrogenase; TERT, telomerase reverse transcriptase; SD rats, Sprague Dawley rats; LVEF, left ventricular ejection fraction; LVFS, left ventricular fractional shortening; LVD, left ventricular diameter; LVAW, left ventricular anterior wall; LVPW, left ventricular posterior wall; LVW, left ventricular weight; NLRP3, NOD-like receptor thermal protein domain associated protein 3; HE, Haematoxylin-Eosin; CD31, platelet endothelial cell adhesion molecule-1; TUNEL, transferase dUTP nick end labelling; SEM, standard error of mean; ANOVA, one-way analysis of variance.
Data Sharing Statement
The datasets and materials used in the study are available from the corresponding author.
Ethics Approval and Consent to Participate
Animal experiments were conducted according to the Guidelines for the Care and Use of Laboratory Animals and were approved by the Ethics Committee of Nanjing Medical University (No. IACUC-2301020).
Author Contributions
All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.
Disclosure
The authors have declared that no competing interest exists in this work.