Nuclear factor erythroid-2 related factor 2 overexpressed mesenchymal stem cells transplantation, improves renal function, decreases injuries markers and increases repair markers in glycerol-induced Acute kidney injury rats

Document Type : Original Article


1 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran

2 Department of Anatomical Sciences and Pathology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

3 School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

4 Department of Medical Laboratory Sciences, Faculty of Medical Sciences, Islamic Azad University, Arak Branch, Arak, Iran


Objective(s):Recently cell therapy is a promising therapeutic modality for many types of disease including acute kidney injury (AKI). Due to the unique biological properties, mesenchymal stem cells (MSCs) are attractive cells in this regard. This study aims to transplant MSCs equipped with nuclear factor E2-related factor 2 (Nrf2) in rat experimental models of acute kidney and evaluate regeneration potential of injured kidney especially expression of injury and repaired biomarkers.
Materials and methods:Nrf2 was overexpressed in bone marrow-derived MSCs by pcDNA.3.1 plasmid. AKI was induced using glycerol in rat models. The regenerative potential of Nrf2-overexpressed MSCs was evaluated in AKI-Induced animal models using biochemical and histological methods after transplantation. Expression of repaired genes, AQP1 and CK-18, as well as injury markers, Kim-1 and Cystatin C, was also assayed in engrafted kidney sections.
Results:Our results revealed that transplantation of Nrf2-overexpressed MSCs into AKI-induced rats decreased blood urea nitrogen and creatinine and ameliorated kidney regeneration throughout 14 days. Upregulation of repaired markers and downregulation of injury markers were considerable 14 days after transplantation. 
Conclusions: Overexpression of Nrf2 in MSCs suggests a new strategy to increase efficiency of MSC-based cell therapy in AKI.


1. Ftouh S, Thomas M. Acute Kidney Injury Guideline Development G. Acute kidney injury: summary of NICE guidance. BMJ 2013; 347:4930-4939.
2. Chertow GM, Burdick E, Honour M, Bonventre JV, Bates DW. Acute kidney injury, mortality, length of stay, and costs in hospitalized patients. JASN 2005; 16:3365-3370.
3. Basile DP, Anderson MD, Sutton TA. Patho-physiology of acute kidney injury. Comprehensive Physiology 2012; 2:1303-1353.
4. Silva Junior GB, Daher Ede F, Mota RM, Menezes FA. Risk factors for death among critically ill patients with acute renal failure. Sao Paulo Med J 2006; 124:257-263.
5. Masoud MS, Anwar SS, Afzal MZ, Mehmood A, Khan SN, Riazuddin S. Pre-conditioned mesenchymal stem cells ameliorate renal ischemic injury in rats by augmented survival and engraftment. J Transl Med 2012; 10:243-254.
6. Humphreys BD, Bonventre JV. Mesenchymal stem cells in acute kidney injury. Ann Rev Med 2008; 59:311-325.
7. Bianchi F, Sala E, Donadei C, Capelli I, La Manna G. Potential advantages of acute kidney injury management by mesenchymal stem cells. World J Stem Cell 2014;6:644-650.
8.Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, et al. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Stem Cell 2008; 26:2075-2082.
9.Doty J, Affleck D, Horne B, Muhlestein J, Psioda M, Paragamian V, et al. A phase 1 trial of human allogeneic mesenchymal stem cells for the prevention of acute kidney injury in cardiac surgery subjects. Am Soc Neph Kidney Week 2011; 8-13.
10.Westenfelder C, Togel FE. Protective actions of administered mesenchymal stem cells in acute kidney injury: relevance to clinical trials. Kidney Int Suppl 2011;1:103-106.
11.Togel F, Cohen A, Zhang P, Yang Y, Hu Z, Westenfelder C. Autologous and allogeneic marrow stromal cells are safe and effective for the treatment of acute kidney injury. Stem Cell Dev 2009; 18:475-485.
12. Amiri F, Jahanian-Najafabadi A, Roudkenar MH.  In vitro augmentation of mesenchymal stem cells viability in stressful microenvironments.  Cell Stress Chaperon 2015; 20:237–251.
13.Mohammadzadeh M, Halabian R, Gharehbaghian A, Amirizadeh N, Jahanian-Najafabadi A, Roushandeh AM, et al. Nrf-2 overexpression in mesenchymal stem cells reduces oxidative stress-induced apoptosis and cytotoxicity. Cell Stress Chaperone 2012; 17:553-565.
14.Lee JM, Li J, Johnson DA, Stein TD, Kraft AD, Calkins MJ, et al. Nrf2, a multi-organ protector? FASEB 2005;19:1061-1066.
15.Kensler TW, Wakabayashi N, Biswal S. Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. Ann Rev Pharmacol Toxicol 2007;47:89-116.
16.Liu M, Grigoryev DN, Crow MT, Haas M, Yamamoto M, Reddy SP, et al. Transcription factor Nrf2 is protective during ischemic and nephrotoxic acute kidney injury in mice. Kidney Int 2009;76:277-285.
17.Ruiz S, Pergola PE, Zager RA, Vaziri ND. Targeting the transcription factor Nrf2 to ameliorate oxidative stress and inflammation in chronic kidney disease. Kidney Int 2013; 83:1029-1041.
18.  Halabian R, Tehrani HA, Jahanian-Najafabadi A, Habibi Roudkenar M. Lipocalin-2-mediated upregulation of various antioxidants and growth factors protects bone marrow-derived mesenchymal stem cells against unfavorable microenvironments. Cell Stress Chaperone 2013; 18: 785-800.
19.Edelstein CL. Biomarkers of acute kidney injury. Adv Chronic Kidney Dis 2008;15:222-234.
20.Li W, Khor TO, Xu C, Shen G, Jeong WS, Yu S, et al. Activation of Nrf2-antioxidant signaling attenuates NFkappaB-inflammatory response and elicits apoptosis. Biochem Pharmacol 2008;76:1485-1489.
21.Bernhardt WM, Campean V, Kany S, Jurgensen JS, Weidemann A, Warnecke C, et al. Preconditional activation of hypoxia-inducible factors ameliorates ischemic acute renal failure. JASN 2006;17:1970-1978.
22.Sohn SJ, Kim SY, Kim HS, Chun YJ, Han SY, Kim SH, et al. In vitro evaluation of biomarkers for cisplatin-induced nephrotoxicity using HK-2 human kidney epithelial cells. Toxicol Lett 2013;217:235-242.
23.Bonventre JV. Kidney injury molecule-1 (KIM-1): a urinary biomarker and much more. Nephrol Dial Transplant 2009;24:3265-3268.
24.Dharnidharka VR, Kwon C, Stevens G. Serum cystatin C is superior to serum creatinine as a marker of kidney function: a meta-analysis. Ame J Kidney Dis 2002;40:221-226.
25.Togel F, Hu Z, Weiss K, Isaac J, Lange C, Westenfelder C. Administered mesenchymal stem cells protect against ischemic acute renal failure through differentiation-independent mechanisms. Ame J Physiol Renal Physiol 2005; 289: 31-42.
26.Togel F, Weiss K, Yang Y, Hu Z, Zhang P, Westenfelder C. Vasculotropic, paracrine actions of infused mesenchymal stem cells are important to the recovery from acute kidney injury. Ame J Physiol Renal Physiol 2007; 292: 1626-1635.
27.Abdel Aziz M, Wassef M, Rashed L, Mhfouz S, Omar N, Elsebaie MM. Mesenchymal stem cells therapy in acute renal failure: possible role of hepatocyte growth factor. J Stem Cell Res Ther 2011; 1:109-120.
28.Deng J, Kohda Y, Chiao H, Wang Y, Hu X, Hewitt SM, et al. Interleukin-10 inhibits ischemic and cisplatin-induced acute renal injury. Kidney Int 2001; 60:2118-2128.
29.Khan M, Akhtar S, Mohsin S, S NK, Riazuddin S. Growth factor preconditioning increases the function of diabetes-impaired mesenchymal stem cells. Stem Cell Dev 2011; 20:67-75.
30. Mohammadzadeh-Vardin M, Habibi Roudkenar M, Jahanian-Najafabadi A. Adenovirus-mediated over-expression of Nrf2 within mesenchymal stem cells (MSCs) protected rats against acute kidney injury. Adv Pharm Bull 2015; 5:201-208.
31. Liu N, Han G, Cheng J, Huang J, Tian J. Erythropoietin promotes the repair effect of acute kidney injury by bone-marrow mesenchymal stem cells transplantation. Exp Biol Med 2013; 238:678-686.
32.  Lee SR, Lee SH, Moon JY, Park JY, Lee D, Lim SJ, et al. Repeated administration of bone marrow-derived mesenchymal stem cells improved the protective effects on a remnant kidney model. Ren Fail 2010; 32:840-848.