1. Micro-vesicles from mesenchymal stem cells over-expressing miR-34a inhibit transforming growth factor-β1-induced epithelial-mesenchymal transition in renal tubular epithelial cells in vitro
- Author
-
Juan He, Ya-Li Jiang, Yan Wang, Xiu-Juan Tian, Shi-Ren Sun, and Li-Min Chen
- Subjects
Male ,Epithelial-Mesenchymal Transition ,Blotting, Western ,Fluorescent Antibody Technique ,lcsh:Medicine ,Cell Line ,Transforming Growth Factor beta1 ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Annexin ,Cricetinae ,medicine ,Animals ,MTT assay ,Epithelial–mesenchymal transition ,Propidium iodide ,Cells, Cultured ,lcsh:R ,Mesenchymal stem cell ,Endoglin ,Cell Differentiation ,Epithelial Cells ,Mesenchymal Stem Cells ,Original Articles ,General Medicine ,Flow Cytometry ,Micro-vesicles ,Molecular biology ,Mice, Inbred C57BL ,MicroRNAs ,Kidney Tubules ,medicine.anatomical_structure ,chemistry ,Cell culture ,030220 oncology & carcinogenesis ,ComputingMethodologies_DOCUMENTANDTEXTPROCESSING ,Bone marrow ,Stem cell ,miR-34a ,030217 neurology & neurosurgery - Abstract
Supplemental Digital Content is available in the text, Background The use of microRNAs in the therapy of kidney disease is hampered by the difficulties in their effective delivery. Micro-vesicles (MVs) are known as natural carriers of small RNAs. Our prior research has demonstrated that MVs isolated from mesenchymal stem cells (MSCs) are capable of attenuating kidney injuries induced by unilateral ureteral obstruction and 5/6 sub-total nephrectomy in mice. The present study aimed to evaluate the effects of miR-34a-5p (miR-34a)-modified MSC-MVs on transforming growth factor (TGF)-β1-induced fibrosis and apoptosis in vitro. Methods Bone marrow MSCs were modified by lentiviruses over-expressing miR-34a, from which MVs were collected for the treatment of human Kidney-2 (HK-2) renal tubular cells exposed to TGF-β1 (6 ng/mL). The survival of HK-2 cells was determined using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and Annexin V-Light 650/propidium iodide (PI) assays. The expression levels of epithelial markers (tight junction protein 1 [TJP1] and E-cadherin) and mesenchymal markers (smooth muscle actin alpha (α-SMA) and fibronectin) in HK-2 cells were measured using Western blot analysis and an immunofluorescence assay. In addition, changes in Notch-1/Jagged-1 signaling were analyzed using Western blotting. Data were analyzed using a Student's t test or one-way analysis of variance. Results MiR-34a expression increased three-fold in MVs generated by miR-34a-modified MSCs compared with that expressed in control MVs (P
- Published
- 2020
- Full Text
- View/download PDF