Induced pluripotent stem cell‐derived conditional medium promotes Leydig cell anti‐apoptosis and proliferation via autophagy and Wnt/β‐catenin pathway

Abstract

Leydig cell transplantation is a better alternative in the treatment of androgen‐deficient males. The main purpose of this study was to investigate the effects of induced pluripotent stem cell‐derived conditioned medium (iPS‐CM) on the anti‐apoptosis, proliferation and function of immature Leydig cells (ILCs), and illuminate the underlying mechanisms. ILCs were exposed to 200 μmol/L hydrogen peroxide (H2O2) for 24 hours with or without iPS‐CM treatments. Cell apoptosis was detected by flow cytometric analysis. Cell proliferation was assessed using cell cycle assays and EdU staining. The steroidogenic enzyme expressions were quantified with Western blotting. The results showed that iPS‐CM significantly reduced H2O2‐induced ILC apoptosis through down‐regulation of autophagic and apoptotic proteins LC3‐I/II, Beclin‐1, P62, P53 and BAX as well as up‐regulation of BCL‐2, which could be inhibited by LY294002 (25 μmol/L). iPS‐CM could also promote ILC proliferation through up‐regulation of β‐catenin and its target proteins cyclin D1, c‐Myc and survivin, but was inhibited by XAV939 (10 μmol/L). The level of bFGF in iPS‐CM was higher than that of DMEM‐LG. Exogenous bFGF (20 ng/mL) or Wnt signalling agonist lithium chloride (LiCl) (20 mmol/L) added into DMEM‐LG could achieve the similar effects of iPS‐CM. Meanwhile, iPS‐CM could improve the medium testosterone levels and up‐regulation of LHCGR, SCARB1, STAR, CYP11A1, HSD3B1, CYP17A1, HSD17B3 and SF‐1 in H2O2‐induced ILCs. In conclusion, iPS‐CM could reduce H2O2‐induced ILC apoptosis through the activation of autophagy, promote proliferation through up‐regulation of Wnt/β‐catenin pathway and enhance testosterone production through increasing steroidogenic enzyme expressions, which might be used in regenerative medicine for future.

The secretome of human dental pulp stem cells protects myoblasts from hypoxia‑induced injury via the Wnt/β‑catenin pathway

“…Moreover, Leroux et al (48) reported that MScs, via the Wnt4 pathway, improved skeletal muscle fiber regeneration following ischemic injury. Other studies have also demonstrated activation of the Wnt pathway by MScs (55)(56)(57). The present study found that, in the presence of hdPSc-cM, the levels of Wnt/β-catenin pathway-related proteins, including p-GSK-3β and β-catenin, were upregulated.…”

RETRACTED: Autophagy regulates the Wnt/GSK3β/β-catenin/cyclin D1 pathway in mesenchymal stem cells (MSCs) exposed to titanium dioxide nanoparticles (TiO2NPs)

“…In our recent study, we observed autophagy induced by TiO 2 NPs in mesenchymal stem cells (MSCs) originating from the bone marrow of humans [ 26 ]. While the role of autophagy in determining cell death or survival is dependent on the internal and external environment and cell type [ 25 ], autophagy has also been known to be related with the Wnt/GSK3β/β-catenin pathway [ 27 ], which is a classic promoter of cell proliferation [ 28 ] and has been regarded as a participant in the maintenance of homeostasis via cell-cell communication [ 29 ].…”

Propofol Inhibits Androgen Production in Rat Immature Leydig Cells

“…Reactive oxidative species (ROS) level was measured with the fluorescence dye 2’7’-dichlorofluorescin diacetate (DCFH-DA) as previously described (Guo et al, 2018). Briefly, 1.5 × 10 5 cells/ml rat immature Leydig cells were seeded into a six-well plate and cultured in 34°C and 5% CO 2 incubator.…”

“…Immature Leydig cells were seeded into a 12-well plate with the density of 5 × 10 5 cells/well and cells were incubated with propofol (0, 5, and 50 µM) for 3 h. To evaluate early and late apoptosis, an Annexin V-FITC/PI Kit was used as previously described (Guo et al, 2018). Cells were harvested and washed with cold PBS and resuspended in 200 µl of Annexin V-binding buffer.…”