Andrographolide protects bone marrow mesenchymal stem cells against glucose and serum deprivation under hypoxia via the NRF2 signaling pathway

Abstract

Background Bone marrow mesenchymal stem cell (BMSCs) therapy is an important cell transplantation strategy in the regenerative medicine field. However, a severely ischemic microenvironment, such as nutrient depletion and hypoxia, causes a lower survival rate of transplanted BMSCs, limiting the application of BMSCs. Therefore, improving BMSCs viability in adverse microenvironments is an important means to improve the effectiveness of BMSCs therapy. Objective To illustrate the protective effect of andrographolide (AG) against glucose and serum deprivation under hypoxia (1% O2) (GSDH)-induced cell injury in BMSCs and investigate the possible underlying mechanisms. Methods An in vitro primary rat BMSCs cell injury model was established by GSDH, and cellular viability, proliferation and apoptosis were observed after AG treatment under GSDH. Reactive oxygen species levels and oxidative stress-related genes and proteins were measured by flow cytometry, RT-qPCR and Western blotting. Mitochondrial morphology, function and number were further assessed by laser confocal microscopy and flow cytometry. Results AG protected BMSCs against GSDH-induced cell injury, as indicated by increases in cell viability and proliferation and mitochondrial number and decreases in apoptosis and oxidative stress. The metabolic status of BMSCs was changed from glycolysis to oxidative phosphorylation to increase the ATP supply. We further observed that the NRF2 pathway was activated by AG, and treatment of BMSCs with a specific NRF2 inhibitor (ML385) blocked the protective effect of AG. Conclusion Our results suggest that AG is a promising agent to improve the therapeutic effect of BMSCs.

Overexpression of PRDX2 in Adipose-Derived Mesenchymal Stem Cells Enhances the Therapeutic Effect in a Neurogenic Erectile Dysfunction Rat Model by Inhibiting Ferroptosis

“…Our results revealed that PRDX2 transfection could increase the proliferation of ADSCs and protect ADSCs from apoptosis and OS in vitro. A possible reason for this is that PRDX2 is an antioxidant enzyme that protects cells, including ADSCs by removing ROS [ 12 , 13 , 25 , 44 , 45 ].…”

“…ADSCs were extracted from epididymal tissue of 4-week-old Sprague-Dawley (SD) male rats [ 24 ]. The cells were cultured in complete medium consisting of low-glucose Dulbecco's modified Eagle's medium (DMEM) (Gibco, USA), 10% fetal bovine serum (FBS) (Gibco, USA), and 1% penicillin-streptomycin solution (containing 100 U/mL penicillin and 100 μ g/mL streptomycin) at 37°C with 5% CO 2 .Then, the ADSCs at passage 3 were collected and incubated with antibodies against CD11b, CD29, CD34, CD44, CD45, and CD90 (all from eBioscience, USA) and detected by flow cytometry (BD FACS Canto, USA) [ 25 ]. As a previous study mentioned [ 24 ], the success of osteogenic and adipogenic differentiation demonstrated the pluripotency of ADSCs (Cyagen, China).…”

Capsaicin acts as a novel NRF2 agonist to suppress ethanol induced gastric mucosa oxidative damage by directly disrupting the KEAP1-NRF2 interaction

“…The relative amount of target gene mRNA was normalized to that of GAPDH mRNA in the same sample. The primers' sequences used in cell experiments were designed through PrimerBank (https://pga.mgh.harvard.edu/primerbank/) and the primers' sequences of PKM2 and LDHA were designed through a reference 63 . The sequence of qPCR primers in the rat experiments were listed using r-ACTB and etc.…”

Study of the effect of keap1 on oxidative stress in human umbilical cord mesenchymal stem cells

“…However, although hucMSCs have achieved good efficacy in clinical trials, we still face great challenges. Differences in oxygen concentration and nutritional conditions between in vivo and in vitro may subject hucMSCs to high oxidative stress after transplantation in vivo, reducing colonization and survival rates and preventing effective treatment of damaged sites [ 35 , 7 , 8 ]. Therefore, it is particularly important to explore the molecular mechanisms that cause oxidative stress in hucMSCs, find intervening targets, and genetically modify MSCs to improve therapeutic efficacy in pediatric diseases.…”