5‐Azacytydine and resveratrol reverse senescence and ageing of adipose stem cells via modulation of mitochondrial dynamics and autophagy

Abstract

Obesity and endocrine disorders have become prevalent issues in the field of both human and veterinary medicine. Equine metabolic syndrome is a complex disorder involving alternation in metabolism and chronic systemic inflammation. It has been shown that unfavourable microenvironment of inflamed adipose tissue negatively affects adipose stem cell population (ASC) residing within, markedly limiting their therapeutic potential. ASCsEMS are characterized by increased senescence apoptosis, excessive accumulation of reactive oxygen species (ROS), mitochondria deterioration and “autophagic flux.” The aim of the present study was to evaluate whether treatment of ASCsEMS with a combination of 5‐azacytydine (AZA) and resveratrol (RES) would reverse aged phenotype of these cells. For this reason, we performed the following analyzes: molecular biology (RT‐PCR), microscopic (immunofluorescence, TEM) and flow cytometry (JC‐1, ROS, Ki67). We evaluated the mitochondrial status, dynamics and clearance as well as autophagic pathways. Furthermore, we investigated epigenetic alternations in treated cells by measuring the expression of TET genes and analysis of DNA methylation status. We have demonstrated that AZA/RES treatment of ASCsEMS is able to rejuvenate these cells by modulating mitochondrial dynamics, in particular by promoting mitochondrial fusion over fission. After AZA/RES treatment, ASCsEMS were characterized by increased proliferation rate, decreased apoptosis and senescence and lower ROS accumulation. Our findings offer a novel approach and potential targets for the beneficial effects of AZA/RES in ameliorating stem cell dysfunctions.

Mitochondrial Dynamics: Fission and Fusion in Fate Determination of Mesenchymal Stem Cells

“…Conversely, cell apoptosis is usually accompanied by abnormal rupture of mitochondria in MSCs. Apoptotic ASCs isolated from equine metabolic syndrome (EMS) horses contain notably increased fragmented mitochondria (Kornicka et al, 2019). Analogously, prominently increased expression of Fis1 and decreased Mfn1 and Mfn2 expression have been reported in apoptotic human umbilical cord MSCs induced by monocrotophos exposure (Srivastava et al, 2018).…”

“…Targeted inhibition of Drp1 with mdivi-1 increases mitochondrial fission and protects human PDLSCs against H 2 O 2 -induced cell dysfunction (He et al, 2018). Mdivi-1 treatment rescues the EMS-induced abnormal mitochondrial network in ASCs, which subsequently reverses the senescence of ASCs, as indicated by suppression of p53, p21, and p62 expression (Kornicka et al, 2019). Moreover, short-term or intermittent administration of mdivi-1 is more beneficial for resisting tissue damage (Jheng et al, 2015).…”

The Effects of Resveratrol in the Treatment of Metabolic Syndrome

“…An in vitro study also showed that resveratrol treatment inhibited preadipocyte proliferation, adipogenic differentiation and inflammatory cytokines production in a Sirt1-dependent manner [77]. Other beneficial effects of resveratrol treatment on adipocytes include improving insulin signaling and mitochondrial dynamics [78,79]. Resveratrol plus quercetin upregulated white adipose tissue SIRT1 and SIRT2 in a rat model of sucrose diet-induced metabolic syndrome [80].…”

Safety of Technique and Procedure of Stromal Vascular Fraction Therapy: From Liposuction to Cell Administration

“…Culturing SVF leads to MSC isolation, and 1-2 week-culture period is needed to produce millions of these fibroblast-like plastic adherent cells known as MSCs [ 6 ]. Autologous SVF extraction procedure replaces any need for allogeneic cell culturing [ 7 ], and therefore risks of contamination, aging of stem cells, or rejection can be avoided [ 8 – 10 ]. Moreover, the procedure is less expensive.…”