Fatima Ali1, Mohsin Khan2, Shaheen N Khan3, Sheikh Riazuddin4. 1. The University of Lahore, Defence Road Campus, Lahore, Pakistan; National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan. Electronic address: Fatemei.ali@gmail.com. 2. National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan; Center for Translational Medicine, Temple University, Lewis Katz School of Medicine, Philadelphia, PA, USA. 3. National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan. 4. The University of Lahore, Defence Road Campus, Lahore, Pakistan; National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan; Allama Iqbal Medical College/Jinnah Hospital Complex, University of Health Sciences, Lahore, Pakistan.
Abstract
BACKGROUND: Stem cell transplantation is one of the therapeutic options available to repair damaged organs. However, transplanted cells entail several challenges including their survival in diabetes-affected injured tissue. This study was designed to determine the effects of preconditioning of mesenchymal stem cells (MSCs) with N-acetyl cysteine (NAC), a widely used antioxidant drug. METHODS: Diabetic-mouse-derived MSCs (blood glucose ≥ 300 mg/dL) were preconditioned with 30 mM NAC for 1 hour followed by oxidative injury with 100 μM hydrogen peroxide (H2O2) for 1 hour. RESULTS: Gene expression analysis showed marked upregulation of prosurvival genes (Akt and Bcl-2) and significantly downregulated expression of proapoptotic and stress genes (Capase-3, Bax, Bak, p53, p38, and NF-κB) in the 30 mM-NAC-treated group when compared with those cells treated with H2O2 alone. NAC preconditioning improved cell viability, decreased lactate dehydrogenase release, β-galactosidase activity, and Annexin-V-positive cells. Also, amelioration of oxidative stress, as shown by a decrease in malondialdehyde level and an increase in superoxide dismutase and catalase activities and glutathione level, was observed in the 30 mM-NAC-treated group in comparison to cells treated with H2O2 alone. CONCLUSION: This study demonstrates the potential benefits of pharmacological preconditioning of diabetic-mouse-derived MSCs with NAC for amelioration of apoptosis and oxidative stress in H2O2 induced injury.
BACKGROUND: Stem cell transplantation is one of the therapeutic options available to repair damaged organs. However, transplanted cells entail several challenges including their survival in diabetes-affected injured tissue. This study was designed to determine the effects of preconditioning of mesenchymal stem cells (MSCs) with N-acetyl cysteine (NAC), a widely used antioxidant drug. METHODS:Diabetic-mouse-derived MSCs (blood glucose ≥ 300 mg/dL) were preconditioned with 30 mM NAC for 1 hour followed by oxidative injury with 100 μM hydrogen peroxide (H2O2) for 1 hour. RESULTS: Gene expression analysis showed marked upregulation of prosurvival genes (Akt and Bcl-2) and significantly downregulated expression of proapoptotic and stress genes (Capase-3, Bax, Bak, p53, p38, and NF-κB) in the 30 mM-NAC-treated group when compared with those cells treated with H2O2 alone. NAC preconditioning improved cell viability, decreased lactate dehydrogenase release, β-galactosidase activity, and Annexin-V-positive cells. Also, amelioration of oxidative stress, as shown by a decrease in malondialdehyde level and an increase in superoxide dismutase and catalase activities and glutathione level, was observed in the 30 mM-NAC-treated group in comparison to cells treated with H2O2 alone. CONCLUSION: This study demonstrates the potential benefits of pharmacological preconditioning of diabetic-mouse-derived MSCs with NAC for amelioration of apoptosis and oxidative stress in H2O2 induced injury.