Research Paper Volume 15, Issue 10 pp 4144—4158
Mechanisms of human umbilical cord mesenchymal stem cells-derived exosomal lncRNA GAS5 in alleviating EMT of HPMCs via Wnt/β-catenin signaling pathway
- 1 Department of Geriatrics, The First Hospital of China Medical University, Shenyang, Liaoning 110001, China
- 2 Department of Nephrology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, China
Received: January 8, 2023 Accepted: May 1, 2023 Published: May 23, 2023
https://doi.org/10.18632/aging.204719How to Cite
Copyright: © 2023 Huang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Background: Prolonged peritoneal dialysis (PD) can result in epithelial-to-mesenchymal transition (EMT) and peritoneal fibrosis (PF), which can cause patients to discontinue PD. It is imperative to urgently investigate effective measures to mitigate PF. This study aims to reveal mechanisms of exosomal lncRNA GAS5 derived from human umbilical cord mesenchymal stem cells (hUC-MSCs) on EMT of human peritoneal mesothelial cells (HPMCs) under high glucose (HG) conditions.Methods: HPMCs were stimulated with 2.5% glucose. The effects on EMT of HPMCs were observed by using an hUC-MSC conditioned medium (hUC-MSC-CM) and extracted exosomes. After hUC-MSCs were transfected with GAS5 siRNA, exosomes were extracted to act on HPMCs for detecting EMT markers, PTEN, and Wnt/β-catenin pathway, lncRNA GAS5 and miR-21 expressions in HPMCs.
Results: We found that HG could induce the EMT of HPMCs. Compared with the HG group, the hUC-MSC-CM could alleviate the EMT of HPMCs induced by HG through exosomes. Exosomes in the hUC-MSC-CM entered HPMCs, by transferring lncRNA GAS5 to HPMCs, which down-regulates miR-21 and up-regulates PTEN, thus finally alleviating EMT of HPMCs. The Wnt/β-catenin pathway plays an essential role in alleviating EMT of HPMCs by exosomes in the hUC-MSC-CM. By transferring lncRNA GAS5 to HPMCs, exosomes derived from hUC-MSCs may competitively bind to miR-21 to regulate suppression on target PTEN genes and alleviate EMT of HPMCs through the Wnt/β-catenin pathway.
Conclusions: Exosomes from the hUC-MSCs-CM could alleviate the EMT of HPMCs induced by HG via regulating lncRNA GAS5/miR-21/PTEN through the Wnt/β-catenin signaling pathway.
Abbreviations
hUC-MSCs: human umbilical cord mesenchymal stem cells; EMT: epithelial-mesenchymal transition; HPMCs: human peritoneal mesothelial cells; HG: high glucose; CM: conditioned medium; hUC-MSC-CM: hUC-MSC conditioned medium; PD: Peritoneal dialysis; ESRD: end-stage renal diseases; PMCs: peritoneal mesothelial cells; UFF: ultrafiltration failure; PF: peritoneal fibrosis; RNAs: ribonucleic acids; DNAs: deoxyribonucleic acids; mRNAs: messenger RNAs; lncRNA: long non-coding RNA; PTEN: phosphatase and tensin homolog deleted on chromosome ten; lncRNA GAS5: lncRNA growth-arrest-specific transcript 5; ceRNA: competing endogenous RNA; DMEM: Dulbecco's modified Eagle medium; NC: negative control; PCR: polymerase chain reaction; NTA: Nanoparticle Tracking Analysis; DAPI: 4’,6-diamidino-2 phenylindole; BCA: bicinchoninic acid; SDS-PAGE: sodium dodecyl sulphate-polyacrylamide gel electrophoresis; PVDF: polyvinylidene fluoride; TBST: Tris-buffered saline Tween; ECL: electrochemiluminescence; cDNAs: complementary DNAs; RT: reverse transcription; PBS: phosphate-buffered saline; EVs: extracellular vesicles; HLSCs: human liver stem-like cells; ADMSC-Exo: exosomes derived from adipose-derived mesenchymal stem cells; TCF/LEF: T-cell factor/lymphoid enhancer factor.