Comparative Clinical Pathology

, Volume 28, Issue 2, pp 311–320 | Cite as

Influence of mesenchymal stem cells and royal jelly on kidney damage triggered by ischemia-reperfusion injury: comparison with ischemic preconditioning in an animal model

  • Gholamreza Didar
  • Fatemeh Delpazir
  • Maryam Kaviani
  • Negar AzarpiraEmail author
  • Lili Sepehrara
  • Padideh Ebadi
  • Farhad Koohpeyma
Original Article


Renal dysfunction due to ischemia/reperfusion (I/R) injury is a common problem following reno-vascular surgery and kidney transplantation. In the present study, we investigated the protective effects of bone marrow mesenchymal stem cells (BM-MSCs), ischemic preconditioning (IPC), and royal jelly (RJ) administration on kidney I/R injury. Male Balb/c mice (30–35 g) were divided randomly into six groups including sham, I/R, I/R after IPC, I/R pre-treated with 100 mg/kg/day RJ for 20 days (I/R + RJ), treatment with 1 × 106 GFP+ BM-MSCs after I/R induction (I/R + MSCs), and I/R pre-treated with 100 mg/kg/day RJ for 20 days and treated with 1 × 106 GFP+ BM-MSCs after I/R induction (I/R + MSCs/RJ). Ten days before beginning the study, the right kidneys of all the mice were removed. I/R injury was induced by clamping the left renal artery for 45 min followed by reperfusion for 48 h. Blood serum was analyzed for blood urea nitrogen and creatinine levels. The histological patterns and the expression of FADD, BAX, BAD, BCL-2, and miR-24 were also evaluated. The BUN level was reduced in I/R + RJ and I/R + MSCs groups significantly. However, the Cr level was significantly decreased just in the I/R + RJ group. The glomerular injury and expression of apoptosis genes were significantly less in the I/R + RJ and I/R + MSCs/RJ groups. The expression of miR-24 showed reduction in all the treated groups unless IPC. Although the previous studies have shown that IPC protects kidney against I/R injury, we found out MSCs and RJ may have a higher potential to protect the kidney tissue. According to the findings, RJ may be effective to protect the kidney against I/R injury.


Ischemia-reperfusion Mesenchymal stem cells Royal Jelly Kidney 



Our special thanks to Dr. Nasrin Shokrpour for the linguistic editing of this manuscript.

Compliance with ethical standards

Our study was compliance with ethical standards.

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution and all applicable institutional guidelines for the care and use of animals were followed.

Informed consent

Humans are not involved in this study.


  1. Alexandropoulos D, Bazigos GV, Doulamis IP, Tzani A, Konstantopoulos P, Tragotsalou N, Kondi-Pafiti A, Kotsis T, Arkadopoulos N, Smyrniotis V, Perrea DN (2017) Protective effects of N-acetylcystein and atorvastatin against renal and hepatic injury in a rat model of intestinal ischemia-reperfusion. Biomed Pharmacother 89:673–680CrossRefGoogle Scholar
  2. Awad AS, El-Sharif AA (2011) Curcumin immune-mediated and anti-apoptotic mechanisms protect against renal ischemia/reperfusion and distant organ induced injuries. Int Immunopharmacol 11:992–996CrossRefGoogle Scholar
  3. Ayupova DA, Singh M, Leonard EC, Basile DP, Lee BS (2009) Expression of the RNA-stabilizing protein HuR in ischemia-reperfusion injury of rat kidney. Am J Physiol Ren Physiol 297:F95–F105CrossRefGoogle Scholar
  4. Azarpira N, Ramzi M, Aghdaie MH, Daraie M, Geramizadeh B (2007) Methylenetetrahydrofolate reductase C677T genotypes and clinical outcome following hematopoietic cell transplant. Exp Clin Transplant 5:693–697Google Scholar
  5. Azarpira N, Ashraf MJ, Aghdaie M (2008a) A 10-year old boy with a hard nodule on his forearm. Archives of Clinical Infectious Diseases 2:99–101Google Scholar
  6. Azarpira N, Ramzi M, Aghdaie MH, Darai M, Geramizadeh B (2008b) Interleukin-10 gene polymorphism in bone marrow transplant recipients. Exp Clin Transplant 6:74–79Google Scholar
  7. Azarpira N, Ramzi M, Aghdaie M, Daraie M (2009) Procalcitonin and C-reactive protein serum levels after hematopoietic stem-cell transplant. Exp Clin Transplant 7:115–118Google Scholar
  8. Azarpira N, Malekhosseini SA, Aghdaie MH, Daraie M (2010) CTLA4 CT60 A/G gene polymorphism in liver transplant recipients. Exp Clin Transplant 8:210–213Google Scholar
  9. Azarpira N, Aghdai MH, Nikeghbalian S, Geramizadeh B, Darai M, Esfandiari E, Bahador A, Kazemi K, Al-Abdullah IH, Malek-Hosseini SA (2014) Human islet cell isolation: the initial step in an islet transplanting program in Shiraz, Southern Iran. Exp Clin Transplant 12:139–142Google Scholar
  10. Badet L, Benhamou PY, Wojtusciszyn A, Baertschiger R, Milliat-Guittard L, Kessler L, Penfornis A, Thivolet C, Renard E, Bosco D (2007) Expectations and strategies regarding islet transplantation: metabolic data from the GRAGIL 2 trial. Transplantation 84:89–96CrossRefGoogle Scholar
  11. Baeyens L, De Breuck S, Lardon J, Mfopou J, Rooman I, Bouwens L (2005) In vitro generation of insulin-producing beta cells from adult exocrine pancreatic cells. Diabetologia 48:49–57CrossRefGoogle Scholar
  12. Bagheri F, Mani A, Tadayyoni A, Firozi F, Nazarinia MA (2013) The prevalence of psychiatric symptoms in the patients with Behcet's disease in Shiraz, Southwest of Iran. J Mood Disorders 3:28CrossRefGoogle Scholar
  13. Baharvand H, Jafary H, Massumi M, Ashtiani SK (2006) Generation of insulin-secreting cells from human embryonic stem cells. Develop Growth Differ 48:323–332CrossRefGoogle Scholar
  14. Bai C, Li X, Gao Y, Wang K, Fan Y, Zhang S, Ma Y, Guan W (2016) Role of microRNA-21 in the formation of insulin-producing cells from pancreatic progenitor cells. Biochim Biophys Acta 1859:280–293CrossRefGoogle Scholar
  15. Banerjee M, Virtanen I, Palgi J, Korsgren O, Otonkoski T (2012) Proliferation and plasticity of human beta cells on physiologically occurring laminin isoforms. Mol Cell Endocrinol 355:78–86CrossRefGoogle Scholar
  16. Banting F, Best CH, Collip JB, Campbell WR, Fletcher AA (1922) Pancreatic extracts in the treatment of diabetes mellitus. Can Med Assoc J 12:141Google Scholar
  17. Baranski JD, Chaturvedi RR, Stevens KR, Eyckmans J, Carvalho B, Solorzano RD, Yang MT, Miller JS, Bhatia SN, Chen CS (2013) Geometric control of vascular networks to enhance engineered tissue integration and function. Proc Natl Acad Sci 110:7586–7591CrossRefGoogle Scholar
  18. Barber FA, Hrnack SA, Snyder SJ, Hapa O (2011) Rotator cuff repair healing influenced by platelet-rich plasma construct augmentation. Arthroscopy 27:1029–1035CrossRefGoogle Scholar
  19. Barker CF, Frangipane LG, Silvers WK (1977) Islet transplantation in genetically determined diabetes. Ann Surg 186:401–410CrossRefGoogle Scholar
  20. Beattie GM, Montgomery AM, Lopez AD, Hao E, Perez B, Just ML, Lakey JR, Hart ME, Hayek A (2002) A novel approach to increase human islet cell mass while preserving β-cell function. Diabetes 51:3435–3439CrossRefGoogle Scholar
  21. Cemek M, Aymelek F, Buyukokuroğlu ME, Karaca T, Buyukben A, Yilmaz F (2010) Protective potential of Royal Jelly against carbon tetrachloride induced-toxicity and changes in the serum sialic acid levels. Food Chem Toxicol 48:2827–2832CrossRefGoogle Scholar
  22. Cho WY, Choi HM, Lee SY, Kim MG, Kim H-K, Jo S-K (2010) The role of Tregs and CD11c+ macrophages/dendritic cells in ischemic preconditioning of the kidney. Kidney Int 78:981–992CrossRefGoogle Scholar
  23. El Morsy EM, Ahmed MAE, Ahmed AAE (2015) Attenuation of renal ischemia/reperfusion injury by açaí extract preconditioning in a rat model. Life Sci 123:35–42CrossRefGoogle Scholar
  24. Endre ZH (2011) Renal ischemic preconditioning: finally some good news for prevention of acute kidney injury. Kidney Int 80:796–798CrossRefGoogle Scholar
  25. Fan R, Yu T, Lin J-L, Ren G-D, Li Y, Liao X-X, Huang Z-T, Jiang C-H (2016) Remote ischemic preconditioning improves post resuscitation cerebral function via overexpressing neuroglobin after cardiac arrest in rats. Brain Res 1648:345–355CrossRefGoogle Scholar
  26. Fontana R, Mendes MA, De Souza BM, Konno K, Cesar LMM, Malaspina O, Palma MS (2004) Jelleines: a family of antimicrobial peptides from the Royal Jelly of honeybees (Apis mellifera). Peptides 25:919–928CrossRefGoogle Scholar
  27. Godwin JG, Ge X, Stephan K, Jurisch A, Tullius SG, Iacomini J (2010) Identification of a microRNA signature of renal ischemia reperfusion injury. Proc Natl Acad Sci 107:14339–14344CrossRefGoogle Scholar
  28. Hernandez DJ, Roberts WB, Miles-Thomas J, Magheli A, Saha S, Schaeffer EM, Racusen LC, Allaf ME (2008) Can ischemic preconditioning ameliorate renal ischemia-reperfusion injury in a single-kidney porcine model? J Endourol 22:2531–2536CrossRefGoogle Scholar
  29. Herrero F, Morales D, Baamonde C, Salas E, Berrazueta JR, Casanova D (2006) Ischemic preconditioning and kidney transplantation: in vivo nitric oxide monitoring in a rat ischemia-reperfusion experimental model. Transplant Proc 38:2600–2602Google Scholar
  30. Joo JD, Kim M, D’agati VD, Lee HT (2006) Ischemic preconditioning provides both acute and delayed protection against renal ischemia and reperfusion injury in mice. J Am Soc Nephrol 17:3115–3123CrossRefGoogle Scholar
  31. Kapitsinou PP, Haase VH (2015) Molecular mechanisms of ischemic preconditioning in the kidney. Am J Physiol Ren Physiol 309:F821–F834CrossRefGoogle Scholar
  32. Karadeniz A, Simsek N, Karakus E, Yildirim S, Kara A, Can I, Kisa F, Emre H, Turkeli M (2011) Royal jelly modulates oxidative stress and apoptosis in liver and kidneys of rats treated with cisplatin. Oxidative Med Cell Longev 2011:1–10CrossRefGoogle Scholar
  33. Kinsey GR, Huang L, Vergis AL, Li L, Okusa MD (2010) Regulatory T cells contribute to the protective effect of ischemic preconditioning in the kidney. Kidney Int 77:771–780CrossRefGoogle Scholar
  34. Kohno K, Okamoto I, Norie A, Iwaki K, Ikeda M, Kurimoto M (2004) Royal jelly inhibits the production of proinflammatory cytokines by activated macrophages. Biosci Biotechnol Biochem 68:138–145CrossRefGoogle Scholar
  35. Kolocassides KG, Galiñanes M, Hearse DJ (1996) Ischemic preconditioning, cardioplegia or both? Differing approaches to myocardial and vascular protection. J Mol Cell Cardiol 28:623–634CrossRefGoogle Scholar
  36. Lee HT, Emala CW (2000) Protective effects of renal ischemic preconditioning and adenosine pretreatment: role of A 1 and A 3 receptors. Am J Physiol Ren Physiol 278:F380–F387CrossRefGoogle Scholar
  37. Lorenzen JM, Kaucsar T, Schauerte C, Schmitt R, Rong S, Hubner A, Scherf K, Fiedler J, Martino F, Kumarswamy R (2014) MicroRNA-24 antagonism prevents renal ischemia reperfusion injury. J Am Soc Nephrol 25:2717–2729CrossRefGoogle Scholar
  38. Mahfoudh-Boussaid A, Zaouali MA, Hadj-Ayed K, Miled A-H, Saidane-Mosbahi D, Rosello-Catafau J, Abdennebi HB (2012) Ischemic preconditioning reduces endoplasmic reticulum stress and upregulates hypoxia inducible factor-1α in ischemic kidney: the role of nitric oxide. J Biomed Sci 19:7CrossRefGoogle Scholar
  39. Nagai T, Inoue R (2004) Preparation and the functional properties of water extract and alkaline extract of royal jelly. Food Chem 84:181–186CrossRefGoogle Scholar
  40. Najafi G, Nejati V, Shalizar Jalali A, Zahmatkesh E (2014) Protective role of royal jelly in oxymetholone-induced oxidative injury in mouse testis. Iran J Toxicol 8:1073–1080Google Scholar
  41. Nowak G, Takacsova-Bakajsova D, Megyesi J (2017) Deletion of protein kinase C-ε attenuates mitochondrial dysfunction and ameliorates ischemic renal injury. Am J Physiol Ren Physiol 312:F109–F120CrossRefGoogle Scholar
  42. Okamoto I, Taniguchi Y, Kunikata T, Kohno K, Iwaki K, Ikeda M, Kurimoto M (2003) Major royal jelly protein 3 modulates immune responses in vitro and in vivo. Life Sci 73:2029–2045CrossRefGoogle Scholar
  43. Park KM, Byun J-Y, Kramers C, Kim JI, Huang PL, Bonventre JV (2003) Inducible nitric-oxide synthase is an important contributor to prolonged protective effects of ischemic preconditioning in the mouse kidney. J Biol Chem 278:27256–27266CrossRefGoogle Scholar
  44. Sadis C, Teske G, Stokman G, Kubjak C, Claessen N, Moore F, Loi P, Diallo B, Barvais L, Goldman M (2007) Nicotine protects kidney from renal ischemia/reperfusion injury through the cholinergic anti-inflammatory pathway. PLoS One 2:e469CrossRefGoogle Scholar
  45. Silici S, Ekmekcioglu O, Kanbur M, Deniz K (2011) The protective effect of royal jelly against cisplatin-induced renal oxidative stress in rats. World J Urol 29:127–132CrossRefGoogle Scholar
  46. Supavekin S, Zhang W, Kucherlapati R, Kaskel FJ, Moore LC, Devarajan P (2003) Differential gene expression following early renal ischemia/reperfusion. Kidney Int 63:1714–1724CrossRefGoogle Scholar
  47. Tian W, Liu Y, Zhang B, Dai X, Li G, Li X, Zhang Z, Du C, Wang H (2015) Infusion of mesenchymal stem cells protects lung transplants from cold ischemia-reperfusion injury in mice. Lung 193:85–95CrossRefGoogle Scholar
  48. Torras J, Herrero-Fresneda I, Lloberas N, Riera M, Cruzado JM, Grinyó JM (2002) Promising effects of ischemic preconditioning in renal transplantation. Kidney Int 61:2218–2227CrossRefGoogle Scholar
  49. Viuda-Martos M, Ruiz-Navajas Y, Fernández-López J, Pérez-Álvarez JA (2008) Functional properties of honey, propolis, and royal jelly. J Food Sci 73:117–124Google Scholar
  50. Vucevic D, Melliou E, Vasilijic S, Gasic S, Ivanovski P, Chinou I, Colic M (2007) Fatty acids isolated from royal jelly modulate dendritic cell-mediated immune response in vitro. Int Immunopharmacol 7:1211–1220CrossRefGoogle Scholar
  51. Yanagita M, Kojima Y, Mori K, Yamada S, Murakami S (2011) Osteoinductive and anti-inflammatory effect of royal jelly on periodontal ligament cells. Biomed Res 32:285–291CrossRefGoogle Scholar
  52. Yoon YE, Lee KS, Choi KH, Kim KH, Yang SC, Han WK (2015) Preconditioning strategies for kidney ischemia reperfusion injury: implications of the “time-window” in remote ischemic preconditioning. PLoS One 10:e0124130CrossRefGoogle Scholar

Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biology, Kazerun BranchIslamic Azad UniversityKazerunIran
  2. 2.Transplant Research CenterShiraz University of Medical SciencesShirazIran
  3. 3.Department of Biochemistry, Faculty of Medicine, Kazerun BranchIslamic Azad UniversityKazerunIran
  4. 4.Shiraz Endocrinology and Metabolism Research CenterShiraz University of Medical SciencesShirazIran

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