Protective effect of gigantol against hydrogen peroxide‑induced apoptosis in rat bone marrow mesenchymal stem cells through the PI3K/Akt pathway

Abstract

Bone marrow mesenchymal stem cell (BMSC) transplants are promising for the treatment of certain central nervous system diseases. However, oxidative stress is one of the major factors that may limit the survival of the transplanted BMSCs. The present study investigated the effect of pretreatment with gigantol on hydrogen peroxide (H2O2)‑induced apoptosis in rat BMSCs (rBMSCs) and the potential underlying mechanisms. The results demonstrated that gigantol pretreatment significantly inhibited H2O2‑induced apoptosis of rBMSCs. rBMSCs were incubated with 600 µM H2O2 in the absence or presence of different doses of gigantol (1‑100 µM). Cell viability and apoptosis ratios were assessed by MTT assays and flow cytometry, respectively. Morphological alterations and reactive oxygen species were measured by the fluorescent‑based methods of Hoechst staining and dichlorodihydrofluorescein diacetate, respectively. Furthermore, the protein levels of phosphorylated‑protein kinase B (Akt), B‑cell lymphoma‑2 (Bcl‑2), Bcl‑2‑associated X (Bax), caspase‑3 and caspase‑9 were investigated by western blotting. Following incubation with H2O2 for 2 h, gigantol significantly inhibited the H2O2‑induced reductions in the cell viability of rBMSCs in a dose‑dependent manner. Furthermore, gigantol upregulated Akt phosphorylation and Bcl‑2 expression, downregulated Bax expression, and reduced the expression of caspase‑3 and caspase‑9 in H2O2‑treated rBMSCs, whereas an opposite effect was detected when LY294002, an inhibitor of phosphatidylinositol 3‑kinase, was administered in combination with gigantol. These results indicate that gigantol may be developed as a promising neuroprotective agent for successful MSC transplantation in ischemic diseases.

Conditioned Medium from Bone Marrow Mesenchymal Stem Cells Restored Oxidative Stress-Related Impaired Osteogenic Differentiation

“…Although the viability of hBMSC was decreased after exposure to the lower concentrations of 125 and 250 µM, these two concentrations had a minimal effect on cellular apoptosis and osteogenic differentiation capacity of hBMSC. In line with our findings, previous study found that the effect of exposure to H 2 O 2 on rat BMSC was also dose-dependent [16]. However, variation among studies in exposure time can contribute to variations in the results, as adverse cellular effects can be induced by lower concentrations of H 2 O 2 when the exposure time is increased [17].…”

“…To mimic the microenvironment of oxidative stress detected in various inflammatory bone diseases and to study the effect of oxidative stress on different cell types in vitro, H 2 O 2 exposure is frequently used as a cellular model of oxidative stress [16][17][18]. Although, at low levels, H 2 O 2 has a beneficial role in basic cellular activity such as proliferation and migration, high levels of H 2 O 2 can cause oxidative stress and lead to adverse effects on cell activity [19,20].…”

H₂O₂ Damages the Stemness of Rat Bone Marrow-Derived Mesenchymal Stem Cells: Developing a “Stemness Loss” Model

“…We found that these 3 characteristics were negatively affected to a different degree when cells were exposed to H 2 O 2 . In our study, we focused on the loss of stemness, a condition of cellular dysfunction (not apoptosis) induced when stem cells are exposed to a concentration of H 2 O 2 below 600 uM [18]. Stemness plays an important role in the ability of stem cells to self-renew [29], to synthesize extracellular matrix components [30,31], and to differentiate [32].…”

“…However, ROS does not always play a damaging role, since recent studies have shown beneficial cell physiological effects at lower concentrations [17]. There have also been many studies investigating methods to protect against oxidative stress and to prevent BMSC apoptosis, such as the use of gigantol [18] or preconditioning with far-infrared irradiation [19].…”

“…However, when the concentration of H 2 O 2 exceeds a certain threshold, BMSC apoptosis can occur. Chen and Huang [18] found that when the concentration of H 2 O 2 reached 600 uM, apoptosis of BMSCs occurred at a relatively high rate. There has been scarce research investigating the effects of H 2 O 2 on the stemness of BMSCs, despite the fact that stemness is a crucial BMSC characteristic when they are going to be transplanted.…”