Nortropane alkaloids as pharmacological chaperones in the rescue of equine adipose-derived mesenchymal stromal stem cells affected by metabolic syndrome through mitochondrial potentiation, endoplasmic reticulum stress mitigation and insulin resistance alleviation

Abstract

Objectives Equine metabolic syndrome (EMS) refers to a cluster of associated abnormalities and metabolic disorders, including insulin resistance and adiposity. The numerous biological properties of mesenchymal stem cells (MSCs), including self-renewal and multipotency, have been the subject of many in-depth studies, for the management of EMS; however, it has been shown that this cell type may be affected by the condition, impairing thus seriously their therapeutic potential. Therefore, an attempt to rescue EMS adipose-derived stem cells (ASCs) with calystegines (polyhydroxylated alkaloids) that are endowed with strong antioxidant and antidiabetic abilities was performed. Methods ASCs isolated from EMS horses were subsequently treated with various concentrations of total calystegines. Different parameters were then assessed using flow cytometry, confocal as well as SE microscopy, and RT-qPCR. Results Our results clearly demonstrated that calystegines could improve EqASC viability and proliferation and significantly reduce apoptosis, via improvement of mitochondrial potentiation and functionality, regulation of pro- and anti-apoptotic pathways, and suppression of ER stress. Furthermore, nortropanes positively upregulated GLUT4 and IRS transcripts, indicating a possible sensitizing or mimetic effect to insulin. Most interesting finding in this investigation lies in the modulatory effect of autophagy, a process that allows the maintenance of cellular homeostasis; calystegines acted as pharmacological chaperones to promote cell survival. Conclusion Obtained data open new perspectives in the development of new drugs, which may improve the metabolic dynamics of cells challenged by MS.

Local, systemic and immunologic safety comparison between xenogeneic equine umbilical cord mesenchymal stem cells, allogeneic canine adipose mesenchymal stem cells and placebo: a randomized controlled trial

“…As positive control, mouse monoclonal anti-equine CD44 (Clone 69-S5, R&D Systems) and mouse monoclonal anti-equine CD90 (Clone MRC OX-7, Abcam) were used at 2 μg/ml. Reactivity of control antibodies against equine antigens was predicted bibliographically ( 39 , 40 ) and confirmed experimentally using several concentrations of antibody on equine MSCs and control cells (negative for CD44 and CD90) (data not shown). After the incubation, wells were washed with 300 μl/well of PBS.…”

Hyoscyamus albus nortropane alkaloids reduce hyperglycemia and hyperinsulinemia induced in HepG2 cells through the regulation of SIRT1/NF-kB/JNK pathway

“…Hyoscyamus albus L is rich in tropane alkaloids, mainly hyoscyamine and scopolamine, which has anticholinergic, analgesic, antispasmodic and sedative properties [ 24 ]. Recently, a new group of polyhydroxylated nortropane alkaloides called calystegines have been found in Hyoscyamus, which sheds a promising light for its application for biomedical application [ 25 , 26 ]. Calystegines as polyhydroxyalkaloids display glycosidase-inhibitory properties by mimicking the pyranosyl or furanosyl moiety of their natural substrates.…”

Astaxanthin Carotenoid Modulates Oxidative Stress in Adipose-Derived Stromal Cells Isolated from Equine Metabolic Syndrome Affected Horses by Targeting Mitochondrial Biogenesis

“…Cultured cells were harvested every three days (80–90% of confluence) and detached from the flasks with a trypsin-EDTA solution (TrypLE Express, Life Technologies; Carlsbad, CA, USA). Cells were multipotent and able to differentiate into adipocytes, chondrocytes and osteoblast in vitro as in previous study [ 53 ].…”