Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway
Abstract Background A growing body of evidence suggests that stem cell-derived exosomal microRNAs (miRNAs) could be a promising cardioprotective therapy in the context of hypoxic conditions. The present study aims to explore how miRNA-144 (miR-144), a miRNA contained in bone marrow mesenchymal stem...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2020-01-01
|
| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-020-1563-8 |
| _version_ | 1830283616388644864 |
|---|---|
| author | Zhuzhi Wen Zun Mai Xiaolin Zhu Tao Wu Yangxin Chen Dengfeng Geng Jingfeng Wang |
| author_facet | Zhuzhi Wen Zun Mai Xiaolin Zhu Tao Wu Yangxin Chen Dengfeng Geng Jingfeng Wang |
| author_sort | Zhuzhi Wen |
| collection | DOAJ |
| description | Abstract Background A growing body of evidence suggests that stem cell-derived exosomal microRNAs (miRNAs) could be a promising cardioprotective therapy in the context of hypoxic conditions. The present study aims to explore how miRNA-144 (miR-144), a miRNA contained in bone marrow mesenchymal stem cell (MSC)-derived exosomes, exerts a cardioprotective effect on cardiomyocyte apoptosis in the context of hypoxic conditions and identify the underlying mechanisms. Methods MSCs were cultured using the whole bone marrow adherent method. MSC-derived exosomes were isolated using the total exosome isolation reagent and confirmed by nanoparticle trafficking analysis as well as western blotting using TSG101 and CD63 as markers. The hypoxic growth conditions for the H9C2 cells were established using the AnaeroPack method. Treatment conditions tested included H9C2 cells pre-incubated with exosomes, transfected with miR-144 mimics or inhibitor, or treated with the PTEN inhibitor SF1670, all under hypoxic growth conditions. Cell apoptosis was determined by flow cytometry using 7-ADD and Annexin V together. The expression levels of the miRNAs were detected by real-time PCR, and the expression levels of AKT/p-AKT, Bcl-2, caspase-3, HIF-1α, PTEN, and Rac-1 were measured by both real-time PCR and western blotting. Results Exosomes were readily internalized by H9C2 cells after co-incubation for 12 h. Exosome-mediated protection of H9C2 cells from apoptosis was accompanied by increasing levels of p-AKT. MiR-144 was found to be highly enriched in MSC-derived exosomes. Transfection of cells with a miR-144 inhibitor weakened exosome-mediated protection from apoptosis. Furthermore, treatment of cells grown in hypoxic conditions with miR-144 mimics resulted in decreased PTEN expression, increased p-AKT expression, and prevented H9C2 cell apoptosis, whereas treatment with a miR-144 inhibitor resulted in increased PTEN expression, decreased p-AKT expression, and enhanced H9C2 cell apoptosis in hypoxic conditions. We also validated that PTEN was a target of miR-144 by using luciferase reporter assay. Additionally, cells treated with SF1670, a PTEN-specific inhibitor, resulted in increased p-AKT expression and decreased H9C2 cell apoptosis. Conclusions These findings demonstrate that MSC-derived exosomes inhibit cell apoptotic injury in hypoxic conditions by delivering miR-144 to cells, where it targets the PTEN/AKT pathway. MSC-derived exosomes could be a promising therapeutic vehicle to facilitate delivery of miRNA therapies to ameliorate ischemic conditions. |
| first_indexed | 2024-12-19T03:00:15Z |
| format | Article |
| id | doaj.art-7e97a48f196941bbadb14a385decd467 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-19T03:00:15Z |
| publishDate | 2020-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-7e97a48f196941bbadb14a385decd4672022-12-21T20:38:13ZengBMCStem Cell Research & Therapy1757-65122020-01-0111111710.1186/s13287-020-1563-8Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathwayZhuzhi Wen0Zun Mai1Xiaolin Zhu2Tao Wu3Yangxin Chen4Dengfeng Geng5Jingfeng Wang6Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen UniversityRNA Biomedical Institute, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityDepartment of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen UniversityDepartment of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen UniversityDepartment of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen UniversityDepartment of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen UniversityDepartment of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen UniversityAbstract Background A growing body of evidence suggests that stem cell-derived exosomal microRNAs (miRNAs) could be a promising cardioprotective therapy in the context of hypoxic conditions. The present study aims to explore how miRNA-144 (miR-144), a miRNA contained in bone marrow mesenchymal stem cell (MSC)-derived exosomes, exerts a cardioprotective effect on cardiomyocyte apoptosis in the context of hypoxic conditions and identify the underlying mechanisms. Methods MSCs were cultured using the whole bone marrow adherent method. MSC-derived exosomes were isolated using the total exosome isolation reagent and confirmed by nanoparticle trafficking analysis as well as western blotting using TSG101 and CD63 as markers. The hypoxic growth conditions for the H9C2 cells were established using the AnaeroPack method. Treatment conditions tested included H9C2 cells pre-incubated with exosomes, transfected with miR-144 mimics or inhibitor, or treated with the PTEN inhibitor SF1670, all under hypoxic growth conditions. Cell apoptosis was determined by flow cytometry using 7-ADD and Annexin V together. The expression levels of the miRNAs were detected by real-time PCR, and the expression levels of AKT/p-AKT, Bcl-2, caspase-3, HIF-1α, PTEN, and Rac-1 were measured by both real-time PCR and western blotting. Results Exosomes were readily internalized by H9C2 cells after co-incubation for 12 h. Exosome-mediated protection of H9C2 cells from apoptosis was accompanied by increasing levels of p-AKT. MiR-144 was found to be highly enriched in MSC-derived exosomes. Transfection of cells with a miR-144 inhibitor weakened exosome-mediated protection from apoptosis. Furthermore, treatment of cells grown in hypoxic conditions with miR-144 mimics resulted in decreased PTEN expression, increased p-AKT expression, and prevented H9C2 cell apoptosis, whereas treatment with a miR-144 inhibitor resulted in increased PTEN expression, decreased p-AKT expression, and enhanced H9C2 cell apoptosis in hypoxic conditions. We also validated that PTEN was a target of miR-144 by using luciferase reporter assay. Additionally, cells treated with SF1670, a PTEN-specific inhibitor, resulted in increased p-AKT expression and decreased H9C2 cell apoptosis. Conclusions These findings demonstrate that MSC-derived exosomes inhibit cell apoptotic injury in hypoxic conditions by delivering miR-144 to cells, where it targets the PTEN/AKT pathway. MSC-derived exosomes could be a promising therapeutic vehicle to facilitate delivery of miRNA therapies to ameliorate ischemic conditions.https://doi.org/10.1186/s13287-020-1563-8Mesenchymal stem cellsExosomesmicroRNA-144PTEN/AKT pathwayCell apoptosis |
| spellingShingle | Zhuzhi Wen Zun Mai Xiaolin Zhu Tao Wu Yangxin Chen Dengfeng Geng Jingfeng Wang Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway Stem Cell Research & Therapy Mesenchymal stem cells Exosomes microRNA-144 PTEN/AKT pathway Cell apoptosis |
| title | Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway |
| title_full | Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway |
| title_fullStr | Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway |
| title_full_unstemmed | Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway |
| title_short | Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway |
| title_sort | mesenchymal stem cell derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microrna144 by targeting the pten akt pathway |
| topic | Mesenchymal stem cells Exosomes microRNA-144 PTEN/AKT pathway Cell apoptosis |
| url | https://doi.org/10.1186/s13287-020-1563-8 |
| work_keys_str_mv | AT zhuzhiwen mesenchymalstemcellderivedexosomesamelioratecardiomyocyteapoptosisinhypoxicconditionsthroughmicrorna144bytargetingtheptenaktpathway AT zunmai mesenchymalstemcellderivedexosomesamelioratecardiomyocyteapoptosisinhypoxicconditionsthroughmicrorna144bytargetingtheptenaktpathway AT xiaolinzhu mesenchymalstemcellderivedexosomesamelioratecardiomyocyteapoptosisinhypoxicconditionsthroughmicrorna144bytargetingtheptenaktpathway AT taowu mesenchymalstemcellderivedexosomesamelioratecardiomyocyteapoptosisinhypoxicconditionsthroughmicrorna144bytargetingtheptenaktpathway AT yangxinchen mesenchymalstemcellderivedexosomesamelioratecardiomyocyteapoptosisinhypoxicconditionsthroughmicrorna144bytargetingtheptenaktpathway AT dengfenggeng mesenchymalstemcellderivedexosomesamelioratecardiomyocyteapoptosisinhypoxicconditionsthroughmicrorna144bytargetingtheptenaktpathway AT jingfengwang mesenchymalstemcellderivedexosomesamelioratecardiomyocyteapoptosisinhypoxicconditionsthroughmicrorna144bytargetingtheptenaktpathway |