International Journal of Stem Cells

Korean Society for Stem Cell Research (한국줄기세포학회)
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Mesenchymal Stem Cells Ameliorate Fibrosis by Enhancing Autophagy via Inhibiting Galectin-3/Akt/mTOR Pathway and by Alleviating the EMT via Inhibiting Galectin-3/Akt/GSK3β/Snail Pathway in NRK-52E Fibrosis

  • Yu Zhao (Department of Human Anatomy and Histoembryology, School of Basic Medical Sciences, Southwest Medical University) ;
  • Chuan Guo (Department of Human Anatomy and Histoembryology, School of Basic Medical Sciences, Southwest Medical University) ;
  • Lianlin Zeng (Department of Human Anatomy and Histoembryology, School of Basic Medical Sciences, Southwest Medical University) ;
  • Jialing Li (Department of Human Anatomy and Histoembryology, School of Basic Medical Sciences, Southwest Medical University) ;
  • Xia Liu (Department of Human Anatomy and Histoembryology, School of Basic Medical Sciences, Southwest Medical University) ;
  • Yiwei Wang (Department of Chemistry, School of Basic Medical Sciences, Southwest Medical University) ;
  • Kun Zhao (Department of Human Anatomy and Histoembryology, School of Basic Medical Sciences, Southwest Medical University) ;
  • Bo Chen (Department of Human Anatomy and Histoembryology, School of Basic Medical Sciences, Southwest Medical University)
  • Received : 2022.01.17
  • Accepted : 2022.02.24
  • Published : 2023.02.28
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Abstract

Background and Objectives: Epithelial-Mesenchymal transition (EMT) is one of the origins of myofibroblasts in renal interstitial fibrosis. Mesenchymal stem cells (MSCs) alleviating EMT has been proved, but the concrete mechanism is unclear. To explore the mechanism, serum-free MSCs conditioned medium (SF-MSCs-CM) was used to treat rat renal tubular epithelial cells (NRK-52E) fibrosis induced by transforming growth factor-β1 (TGF-β1) which ameliorated EMT. Methods and Results: Galectin-3 knockdown (Gal-3 KD) and overexpression (Gal-3 OE) lentiviral vectors were established and transfected into NRK-52E. NRK-52E fibrosis model was induced by TGF-β1 and treated with the SF-MSCs-CM for 24 h after modelling. Fibrosis and autophagy related indexes were detected by western blot and immunocytochemistry. In model group, the expressions of α-smooth muscle actin (α-SMA), fibronectin (FN), Galectin-3, Snail, Kim-1, and the ratios of P-Akt/Akt, P-GSK3β/GSK3β, P-PI3K/PI3K, P-mTOR/mTOR, TIMP1/MMP9, and LC3B-II/I were obviously increased, and E-Cadherin (E-cad) and P62 decreased significantly compared with control group. SF-MSCs-CM showed an opposite trend after treatment compared with model group. Whether in Gal-3 KD or Gal-3 OE NRK-52E cells, SF-MSCs-CM also showed similar trends. However, the effects of anti-fibrosis and enhanced autophagy in Gal-3 KD cells were more obvious than those in Gal-3 OE cells. Conclusions: SF-MSCs-CM probably alleviated the EMT via inhibiting Galectin-3/Akt/GSK3β/Snail pathway. Meanwhile, Gal-3 KD possibly enhanced autophagy via inhibiting Galectin-3/Akt/mTOR pathway, which synergistically ameliorated renal fibrosis. Targeting galectin-3 may be a potential target for the treatment of renal fibrosis.

Keywords

Acknowledgement

We thank the Beijing Syngentech Co., LTD, for the support for lentivirus transfection technology. This work was supported by the Science and Technology Department of Sichuan Province (No.2018JY0490), A Project Supported by Scientific Research Fund of Sichuan Provincial Education Department (No. 18ZA0524), Research project of Sichuan Traditional Chinese Medicine Administration (2021MS553), the Health and Family Planning Commission of Sichuan province (No. 16PJ540), Luzhou Science and Technology Bureau (No.2016-S-65(1/9), 2016LZXNYD-J18), and National Innovation and Entrepreneurship Program for College Students (S202110632020; S202110632031).

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