Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in Podocytes
Diabetic nephropathy is a major complication in diabetic patients. Podocytes undergo loss and detachment from the basal membrane. Intra- and intercellular communication through exosomes are key processes for maintaining function, and the Rab3A/Rab27A system is an important counterpart. Previously, w...
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| Format: | Article |
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MDPI AG
2023-05-01
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| Series: | Biology |
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| Online Access: | https://www.mdpi.com/2079-7737/12/5/690 |
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| author | Olga Martinez-Arroyo Ana Flores-Chova Belen Sanchez-Garcia Josep Redon Raquel Cortes Ana Ortega |
| author_facet | Olga Martinez-Arroyo Ana Flores-Chova Belen Sanchez-Garcia Josep Redon Raquel Cortes Ana Ortega |
| author_sort | Olga Martinez-Arroyo |
| collection | DOAJ |
| description | Diabetic nephropathy is a major complication in diabetic patients. Podocytes undergo loss and detachment from the basal membrane. Intra- and intercellular communication through exosomes are key processes for maintaining function, and the Rab3A/Rab27A system is an important counterpart. Previously, we observed significant changes in the Rab3A/Rab27A system in podocytes under glucose overload, demonstrating its important role in podocyte injury. We investigated the implication of silencing the Rab3A/Rab27A system in high glucose-treated podocytes and analysed the effect on differentiation, apoptosis, cytoskeletal organisation, vesicle distribution, and microRNA expression in cells and exosomes. For this, we subjected podocytes to high glucose and transfection through siRNAs, and we isolated extracellular vesicles and performed western blotting, transmission electron microscopy, RT-qPCR, immunofluorescence and flow cytometry assays. We found that silencing <i>RAB3A</i> and <i>RAB27A</i> generally leads to a decrease in podocyte differentiation and cytoskeleton organization and an increase in apoptosis. Moreover, CD63-positive vesicles experienced a pattern distribution change. Under high glucose, Rab3A/Rab27A silencing ameliorates some of these detrimental processes, suggesting a differential influence depending on the presence or absence of cellular stress. We also observed substantial expression changes in miRNAs that were relevant in diabetic nephropathy upon silencing and glucose treatment. Our findings highlight the Rab3A/Rab27A system as a key participant in podocyte injury and vesicular traffic regulation in diabetic nephropathy. |
| first_indexed | 2024-03-11T03:55:36Z |
| format | Article |
| id | doaj.art-50b3cab9b1dd40cfa261fc982df62713 |
| institution | Directory Open Access Journal |
| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-11T03:55:36Z |
| publishDate | 2023-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biology |
| spelling | doaj.art-50b3cab9b1dd40cfa261fc982df627132023-11-18T00:33:01ZengMDPI AGBiology2079-77372023-05-0112569010.3390/biology12050690Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in PodocytesOlga Martinez-Arroyo0Ana Flores-Chova1Belen Sanchez-Garcia2Josep Redon3Raquel Cortes4Ana Ortega5Cardiometabolic and Renal Risk Research Group, Biomedical Research Institute of Hospital Clinico de Valencia INCLIVA, 46010 Valencia, SpainCardiometabolic and Renal Risk Research Group, Biomedical Research Institute of Hospital Clinico de Valencia INCLIVA, 46010 Valencia, SpainCardiometabolic and Renal Risk Research Group, Biomedical Research Institute of Hospital Clinico de Valencia INCLIVA, 46010 Valencia, SpainCardiometabolic and Renal Risk Research Group, Biomedical Research Institute of Hospital Clinico de Valencia INCLIVA, 46010 Valencia, SpainCardiometabolic and Renal Risk Research Group, Biomedical Research Institute of Hospital Clinico de Valencia INCLIVA, 46010 Valencia, SpainCardiometabolic and Renal Risk Research Group, Biomedical Research Institute of Hospital Clinico de Valencia INCLIVA, 46010 Valencia, SpainDiabetic nephropathy is a major complication in diabetic patients. Podocytes undergo loss and detachment from the basal membrane. Intra- and intercellular communication through exosomes are key processes for maintaining function, and the Rab3A/Rab27A system is an important counterpart. Previously, we observed significant changes in the Rab3A/Rab27A system in podocytes under glucose overload, demonstrating its important role in podocyte injury. We investigated the implication of silencing the Rab3A/Rab27A system in high glucose-treated podocytes and analysed the effect on differentiation, apoptosis, cytoskeletal organisation, vesicle distribution, and microRNA expression in cells and exosomes. For this, we subjected podocytes to high glucose and transfection through siRNAs, and we isolated extracellular vesicles and performed western blotting, transmission electron microscopy, RT-qPCR, immunofluorescence and flow cytometry assays. We found that silencing <i>RAB3A</i> and <i>RAB27A</i> generally leads to a decrease in podocyte differentiation and cytoskeleton organization and an increase in apoptosis. Moreover, CD63-positive vesicles experienced a pattern distribution change. Under high glucose, Rab3A/Rab27A silencing ameliorates some of these detrimental processes, suggesting a differential influence depending on the presence or absence of cellular stress. We also observed substantial expression changes in miRNAs that were relevant in diabetic nephropathy upon silencing and glucose treatment. Our findings highlight the Rab3A/Rab27A system as a key participant in podocyte injury and vesicular traffic regulation in diabetic nephropathy.https://www.mdpi.com/2079-7737/12/5/690diabetic nephropathygene silencingpodocyteRab3A/Rab27A systemvesicular traffic |
| spellingShingle | Olga Martinez-Arroyo Ana Flores-Chova Belen Sanchez-Garcia Josep Redon Raquel Cortes Ana Ortega Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in Podocytes Biology diabetic nephropathy gene silencing podocyte Rab3A/Rab27A system vesicular traffic |
| title | Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in Podocytes |
| title_full | Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in Podocytes |
| title_fullStr | Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in Podocytes |
| title_full_unstemmed | Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in Podocytes |
| title_short | Rab3A/Rab27A System Silencing Ameliorates High Glucose-Induced Injury in Podocytes |
| title_sort | rab3a rab27a system silencing ameliorates high glucose induced injury in podocytes |
| topic | diabetic nephropathy gene silencing podocyte Rab3A/Rab27A system vesicular traffic |
| url | https://www.mdpi.com/2079-7737/12/5/690 |
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