Immunomodulatory Properties of Adipose-Derived Stem Cells Treated with 5-Azacytydine and Resveratrol on Peripheral Blood Mononuclear Cells and Macrophages in Metabolic Syndrome Animals.

Katarzyna Kornicka,Krzysztof Marycz,Agnieszka Śmieszek,Agnieszka Sławomira Węgrzyn,Michael Röcken

doi:10.3390/jcm7110383

Abstract

Endocrine disorders, including equine metabolic syndrome (EMS), are a serious issue in veterinary medicine and horse breeding. Furthermore, EMS was shown to affect the cytophysiological properties of adipose-derived stem cells, reducing their therapeutic potential. However, it was shown that those cells can be rejuvenated while using a combination of two chemicals: 5-azacytydine (AZA) and resveratrol (RES). In the present study, we decided to evaluate the immunomodulatory properties of AZA/RES-treated adipose-derived stem cells (ASC) isolated from EMS horses (ASCEMS). Thus, we co-cultured ASC with peripheral blood mononuclear cells (PBMC) and RAW264.7 macrophages. Most attention was placed on regulatory T lymphocytes (TREG), as well as the messenger RNA (mRNA) and protein levels of several cytokines (tumor necrosis factor α (TNF-α), interleukin (IL)-6, IL-10, and IL-1β). Moreover, we also investigated the expression of genes related to auto- and mitophagy in both PBMCs and ASCs. PBMCs were obtained from healthy and EMS-suffering individuals and were co-cultured with ASCs that were isolated from healthy and EMS horses cultured in control conditions and with AZA/RES. We discovered that cells treated with AZA/RES increase the TREG number while co-cultured with PBMCs. Moreover, the co-culture of PBMCs with AZA/RES-treated ASCEMS induced mitophagy in PBMCs. Furthermore, ASCEMS pre-treated with AZA/RES displayed anti-inflammatory properties, as decreased levels of TNF-α, nitric oxide (NO), and IL-6 were observed in those cells in comparison with their untreated counterparts in the co-culture with RAW264.7 macrophages. In summary, we demonstrated that ASCEMS treated with AZA/RES displayed increased anti-inflammatory properties, and was able to regulate and activate the TREG-related anti-inflammatory response.

Highlights

Mesenchymal stem cells (MSCs) are a progenitor stem-cell population isolated from multiple tissues, mainly bone marrow (BMSC) and adipose tissue (ASC)
In our previous study, we reported that the treatment of ASCs from EMS (ASCEMS) with AZA/RES reversed their aged phenotype and rejuvenated them, so that their physiological properties were comparable to those observed in ASCCTRL
We have found for the first time that ASCEMS and ASCEMS that are treated with AZA/RES induced and increased Treg activity and caused dynamic alternations in the cytokine expression pattern

Summary

Introduction

Mesenchymal stem cells (MSCs) are a progenitor stem-cell population isolated from multiple tissues, mainly bone marrow (BMSC) and adipose tissue (ASC). MSCs are characterized by low immunogenicity and immunoregulatory properties [1] For these reasons, the application of MSCs in the treatment of autoimmune disorders and tissue damage, and in the induction of allogenic transplant tolerance is fully reasonable and promising [2,3]. Multiple studies indicated that MSCs regulate T-cell and B-cell functions They suppress T-cell proliferation and cytokine secretion, and they regulate the T helper 1/T helper 2 (Th1/Th2) cell balance [6,7]. They are able to regulate the functions of regulatory T cells (TREG) [8], which indicates that MSCs may represent an effective therapeutic strategy for metabolic syndrome treatment

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Results

Conclusion