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Cited by 146Extracellular vesicles in diseaseCardiac Ischemia and ReperfusionMicroRNA in disease regulation

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Bone marrow mesenchymal stem cell-secreted exosomes carrying microRNA-125b protect against myocardial ischemia reperfusion injury via targeting SIRT7

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Chen, Qi, Liu, Yu, Ding, Xueyan, Li, Qinfeng, Qiu, Fuyu, Wang, Meihui, Shen, Zhida, Zheng, Hao, Fu, Guosheng·2020·Molecular and Cellular Biochemistry
DOI 10.1007/s11010-019-03671-zPMC PMC6955239
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Abstract

MicroRNA-125b (miR-125b) reduces myocardial infarct area and restrains myocardial ischemia reperfusion injury (I/R). In this study, we aimed to investigate the effect of bone marrow mesenchymal stem cell (BMSC)-derived exosomes carrying miR-125b on I/R rats. The myocardial I/R model in rats was constructed by ligation of the left anterior descending coronary artery (LAD). Rats were randomly divided into I/R and Sham group. Lv-cel-miR-67 (control) or Lv-miR-125b was transfected into BMSCs. Exosomes were extracted from transfected BMSCs, and separately named BMSC-Exo-67, BMSC-Exo-125b, and BMSC-Exo. MTT assay and flow cytometry were used to detect the viability and apoptosis of I/R myocardium cells, respectively. The expression of cell apoptosis proteins and the levels of inflammatory factors were examined by Western blot and ELISA assay, respectively. The target relationship between miR-125b and SIRT7 was predicted by using StarBase3.0, and was confirmed by using dual-luciferase reporter gene assay. qRT-PCR, immunohistochemistry staining, and Western blot were used to evaluate the expression of SIRT7 in myocardium tissues in I/R rats. BMSC-derived exosomes were successfully isolated and identified by TEM and positive expression of CD9 and CD63. The expression of miR-125b was down-regulated in I/R myocardium tissues and cells. BMSC-Exo-125b significantly up-regulated miR-125b in I/R myocardium cells. The intervention of BMSC-Exo-125b significantly increased the cell viability, decreased the apoptotic ratio, down-regulated Bax and caspase-3, up-regulated Bcl-2, and decreased the levels of IL-1β, IL-6, and TNF-α in I/R myocardium cells. SIRT7 was a target of miR-125b, and BMSC-Exo-125b significantly down-regulated SIRT7 in myocardium cells. In addition, the injection of BMSC-Exo-125b alleviated the pathological damages and down-regulated SIRT7 in myocardium tissues of I/R rats. BMSC-derived exosomes carrying miR-125b protected against myocardial I/R by targeting SIRT7.

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Fig. 1 Characterization of exosomes derived from bone marrow mesenchymal stem cells (BMSCs). a Cellular morphology (P1, P3) of BMSCs observed under an inverted fluorescence microscope. Scale bar: 100 μm. b Flow cytometry was used to analyze the surface antigens (CD44, CD105, CD31) in BMSCs. c, d The morphology of FBS-derived exosomes and BMSC-derived exosomes was observed under transmission electron microscopy (TEM). Scale bar: 50 μm. e Western blot was used to examine the expression of CD9, CD63 in BMSCs, and BMSC-derived exosomes
Fig. 1 Characterization of exosomes derived from bone marrow mesenchymal stem cells (BMSCs). a Cellular morphology (P1, P3) of BMSCs observed under an inverted fluorescence microscope. Scale bar: 100 μm. b Flow cytometry was used to analyze the surface antigens (CD44, CD105, CD31) in BMSCs. c, d The morphology of FBS-derived exosomes and BMSC-derived exosomes was observed under transmission electron microscopy (TEM). Scale bar: 50 μm. e Western blot was used to examine the expression of CD9, CD63 in BMSCs, and BMSC-derived exosomesCC BY · PubMed Central
Fig. 2 Myocardial ischemia reperfusion injury (I/R) model in rats. a Hemodynamic test of I/R rats. b Hematoxylin–eosin (HE) staining was performed to detect the pathological changes of myocardium in I/R rats (× 400). c Triphenyltetrazolium chloride (TTC) assay was used to evaluate the infarct size in I/R rats
Fig. 2 Myocardial ischemia reperfusion injury (I/R) model in rats. a Hemodynamic test of I/R rats. b Hematoxylin–eosin (HE) staining was performed to detect the pathological changes of myocardium in I/R rats (× 400). c Triphenyltetrazolium chloride (TTC) assay was used to evaluate the infarct size in I/R ratsCC BY · PubMed Central
Fig. 3 Bone marrow mesenchymal stem cells (BMSCs)-derived exosomes increased the expression of miR-125b in I/R myocardium cells. a, b The relative expression of miR-125b was examined by qRT-PCR in myocardium tissues and cells. c, d The relative expression of miR-125b was detected by qRT-PCR in BMSC cells and BMSC-derived exosomes, respectively. e BMSC-Exo-125b was traced by labeling PKH26 (red). f The relative expression of miR-125b was examined by qRT-PCR in I/R myocardium cells. (Color figure online)
Fig. 3 Bone marrow mesenchymal stem cells (BMSCs)-derived exosomes increased the expression of miR-125b in I/R myocardium cells. a, b The relative expression of miR-125b was examined by qRT-PCR in myocardium tissues and cells. c, d The relative expression of miR-125b was detected by qRT-PCR in BMSC cells and BMSC-derived exosomes, respectively. e BMSC-Exo-125b was traced by labeling PKH26 (red). f The relative expression of miR-125b was examined by qRT-PCR in I/R myocardium cells. (Color figure online)CC BY · PubMed Central
Fig. 4 BMSC-Exo-125b inhibited the apoptosis of ischemia reperfusion injury (I/R) myocardium cells. a The viability of myocardium cells was examined by MTT. b Apoptotic ratio (%, cell ratio in LR + UR) of myocardium cells was examined by flow cytometry. Lower left quadrant (LL), viable cells; upper left quadrant (UL), necrotic cells; lower right quadrant (LR), early apoptotic cells; upper right quadrant (UR), late apoptotic cells. c Western blot was used to detect the expression of apoptotic proteins in myocardium cells. d ELISA was performed to examine the levels of inflammatory factors
Fig. 4 BMSC-Exo-125b inhibited the apoptosis of ischemia reperfusion injury (I/R) myocardium cells. a The viability of myocardium cells was examined by MTT. b Apoptotic ratio (%, cell ratio in LR + UR) of myocardium cells was examined by flow cytometry. Lower left quadrant (LL), viable cells; upper left quadrant (UL), necrotic cells; lower right quadrant (LR), early apoptotic cells; upper right quadrant (UR), late apoptotic cells. c Western blot was used to detect the expression of apoptotic proteins in myocardium cells. d ELISA was performed to examine the levels of inflammatory factorsCC BY · PubMed Central
Fig. 5 MiR-125b was negatively correlated with SIRT7. a A binding site at 3′-UTR of SIRT7 was predicted on miR-125b by StarBase3.0. b The luciferase activity was measured by dual-luciferase reporter gene assay. c Relative mRNA expression of SIRT7 in ischemia reperfusion injury (I/R) myocardium tissues was examined by qRT-PCR. d Spearman’s correlation analysis was used to detect the correlation between miR-125b and SIRT7 expression. e, f qRT-PCR and Western blot were used to detect the mRNA and protein expression of SIRT7 in myocardium cells
Fig. 5 MiR-125b was negatively correlated with SIRT7. a A binding site at 3′-UTR of SIRT7 was predicted on miR-125b by StarBase3.0. b The luciferase activity was measured by dual-luciferase reporter gene assay. c Relative mRNA expression of SIRT7 in ischemia reperfusion injury (I/R) myocardium tissues was examined by qRT-PCR. d Spearman’s correlation analysis was used to detect the correlation between miR-125b and SIRT7 expression. e, f qRT-PCR and Western blot were used to detect the mRNA and protein expression of SIRT7 in myocardium cellsCC BY · PubMed Central
Fig. 6 BMSC-Exo-125b restored the cardiac function of myocardial ischemia reperfusion injury (I/R) rats. a Hemodynamic test of I/R rats. b HE staining was performed to detect the pathological changes of myocardium in I/R rats (× 400). c Triphenyltetrazolium chloride (TTC) assay was used to evaluate the infarct size in I/R rats. d qRT-PCR was used to examine the relative mRNA expression of SIRT7 in myocardium tissues. e Relative density of SIRT7 was detected by immunohistochemistry in myocardium tissues (× 400). f Western blot was used to examine the relative protein expression of SIRT7 in myocardium tissues
Fig. 6 BMSC-Exo-125b restored the cardiac function of myocardial ischemia reperfusion injury (I/R) rats. a Hemodynamic test of I/R rats. b HE staining was performed to detect the pathological changes of myocardium in I/R rats (× 400). c Triphenyltetrazolium chloride (TTC) assay was used to evaluate the infarct size in I/R rats. d qRT-PCR was used to examine the relative mRNA expression of SIRT7 in myocardium tissues. e Relative density of SIRT7 was detected by immunohistochemistry in myocardium tissues (× 400). f Western blot was used to examine the relative protein expression of SIRT7 in myocardium tissuesCC BY · PubMed Central

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Metadata

Authors
Chen, Q., Liu, Y., Ding, X., Li, Q., Qiu, F., Wang, M., Shen, Z., Zheng, H., & Fu, G.
Affiliations
  • Sir Run Run Shaw Hospital (CN)
  • Nanjing Drum Tower Hospital (CN)
  • Zhejiang Provincial People's Hospital (CN)
Corresponding author
  • Zheng, Hao
  • Fu, Guosheng
ORCID
Venue
Molecular and Cellular Biochemistry
Publisher
Springer Science and Business Media LLC
Published
2019-12-19
Volume
465
Issue
1-2
Pages
103-114
ISSN
  • 0300-8177 (ISSN-L)
  • 1573-4919
DOI
10.1007/s11010-019-03671-z
PMC
PMC6955239
OpenAlex
W2996170603
Cited by
146
References
47
Citations by year
2026: 12 · 2025: 19 · 2024: 24 · 2023: 26 · 2022: 30 · 2021: 26 · 2020: 9
FWCI
5.37
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular and Cellular Biochemistry (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Effects and mechanisms of microRNA and O-GlcNAcylation on paracrine function of diabetic endothelial progenitor cell (Y18H020004)
  • The mechanism research identifing Oct4 upregulating Cripto-1 mediated osteoblast differentiation of valve interstitial cells during the pathogenesis of aortic valve calcification (81800342)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiac Ischemia and Reperfusion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
Keywords
bone marrow mesenchymal stem cells, exosomes, miR-125b, SIRT7, myocardial ischemia-reperfusion injury, cardioprotection, apoptosis, inflammation, stem cell-derived exosomes, myocardial infarction, Bax, caspase-3, Bcl-2, inflammatory cytokines, microRNA target regulation, exosome delivery
MeSH terms
Animals, Bone Marrow Cells, Male, Myocardial Reperfusion Injury, Cytokines, Rats, Sprague-Dawley, Apoptosis, MicroRNAs, Sirtuins, Rats, Exosomes, Mesenchymal Stem Cells

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