Modulation of inflammatory pathways in ischemic stroke rats by hypoxia-primed umbilical cord mesenchymal stem cells (UC-MSCs): Implications for IFN-γ and IL-10 signaling

  • Rakha Fahreza Graduate Student of Biomedical Sciences Master Program, Faculty of Medicine, Sultan Agung Islamic University, Semarang, Indonesia
  • Eko Setiawan Biomedical Sciences Master Program, Faculty of Medicine, Sultan Agung Islamic University, Semarang, Indonesia
  • Setyo Trisnadi Biomedical Sciences Master Program, Faculty of Medicine, Sultan Agung Islamic University, Semarang, Indonesia
  • Agung Putra Biomedical Sciences Doctoral Program, Faculty of Medicine, Sultan Agung Islamic University, Semarang, Indonesia/Stem Cell and Cancer Research Laboratory, Semarang, Indonesia
  • Nurul Hidayah Stem Cell and Cancer Research Laboratory, Semarang, Indonesia/Biotechnology Undergraduate Program, Institut Karya Mulia Bangsa, Semarang, Indonesia
  • Fikriya Novita Sari Stem Cell and Cancer Research Laboratory, Semarang, Indonesia
  • Sugeng Ibrahim Faculty of Medicine, Universitas Islam Negeri Walisongo, Semarang, Indonesia
Keywords: ischemic stroke, HUC-MSCs, hypoxia, IFN-γ, IL-10

Abstract

Ischemic stroke is a major global cause of disability and mortality. Inflammation plays a central role in its pathogenesis, characterized by elevated pro-inflammatory cytokines such as IFN-γ and reduced anti-inflammatory cytokines like IL-10. Mesenchymal stem cells (MSCs), especially those derived from the umbilical cord (UC-MSCs), exhibit enhanced immunomodulatory potential when preconditioned under hypoxia. This study aims to evaluate the effect of hypoxic-preconditioned UC-MSCs administration on IFN-γ and IL-10 levels in an ischemic stroke rat model. This in vivo experimental study employed a randomized posttest-only control group design with four groups of male Wistar rats (n=6 each), ranging from healthy controls, untreated group to stroke-induced groups treated with hypoxic UC-MSCs at different doses (1.5x106 and 3x106). IFN-γ and IL-10 levels in brain tissue of each group were measured via ELISA. Significant reduction in IFN-γ and elevation in IL-10 were observed in UC-MSC-treated groups, particularly at the 3×10⁶ cell dose compared to the untreated ischemic group (p<0.05). Hypoxic UC-MSCs reduce post-stroke inflammation by lowering IFN-γ and enhancing IL-10, indicating a promising immunomodulatory potential.

References

Boshuizen MCS, Steinberg GK. Stem cell-based immunomodulation after stroke: effects on brain repair processes. Stroke 2018; 49(6):1563-70.

https://doi.org/10.1161/STROKEAHA.117.020465

Choudhery MS, Arif T, Mahmood R, Harris DT. Therapeutic potential of mesenchymal stem cells in stroke: mechanisms and prospects. Biomolecules 2025; 15(4):558.

https://doi.org/10.3390/biomolecules15040558

Cunningham CJ, Redondo-Castro E, Allan SM. The therapeutic potential of the mesenchymal stem cell secretome in ischeamic stroke. J Cereb Blood Flow Metab 2018; 38(8):1276-92.

https://doi.org/10.1177/0271678X18776802

De Palma ST, Hermans EC, Shamorkina TM, Trayford C, van Rijt S, Heck AJR, et al. Hypoxic preconditioning enhances the potential of MSC therapy in neonatal Hypoxic-ischemic brain injury. Stroke 2025; 56(7):1872-82.

https://doi.org/10.1161/STROKEAHA.124.048964

Lee BC, Kang KS. Functional enhancement strategies for immunomodulation of mesenchymal stem cells and their therapeutic application. Stem Cell Res Ther 2020; 11(1):397.

https://doi.org/10.1186/s13287-020-01920-3

Lin YC, Ko TL, Shih YH, Lin MYA, Fu TW, Hsiao HS, et al. Human umbilical mesenchymal stem cells promote recovery after ischemic stroke. Stroke 201; 42(7):2045-53.

https://doi.org/10.1161/STROKEAHA.110.603621

Noronha NC, Mizukami A, Caliári-Oliveira C, Cominal JG, Rocha JLM, Covas DT, et al. Priming approaches to improve the efficacy of mesenchymal stromal cell-based therapy. Stem Cells Res Ther 2019; 10(1):131.

https://doi.org/10.1186/s13287-019-1224-y

Moeinabadi-Bidgoli K, Babajani A, Yazdanpanah G, Farhadihosseinabadi B, Jamshidi E, Bahrami S, et al. Translational insights into stem cell preconditioning: From molecular mechanisms to preclinical applications. Biomed Pharmacother 2021; 142:112026.

https://doi.org/10.1016/j.biopha.2021.112026

Sarsenova M, Kim Y, Raziyeva K, Kazybay B, Ogay V, Saparov A. Recent advances to enhance the immunomodulatory capacity of MSC-based therapies. Front Immunol 2022; 13:1010399.

https://doi.org/10.3389/fimmu.2022.1010399

Shen D, Wang H, Zhu H, Jiang C, Xie F, Zhang H, et al. Pre-clinical efficacy evaluation of human umbilical cord mesenchymal stem cells for ischemic stroke. Front Immunol 2023; 13:1095469.

https://doi.org/10.3389/fimmu.2022.1095469

Stonesifer C, Corey S, Ghanekar S, Diamandis Z, Acosta SA, Borlongan CV. Stem cell therapy for abrogating stroke-induced neuroinflammation and relevant secondary cell death mechanisms. Prog Neurobiol 2017; 158:94-131.

https://doi.org/10.1016/j.pneurobio.2017.07.004

Tan YL, Al-Masawa ME, Eng SP, Shafiee MN, Law JX, Ng MH. Therapeutic efficacy of interferon-gamma and hypoxia-primed mesenchymal stromal cells and their extracellular vesicles: underlying mechanisms and potentials in clinical translation. Biomedicines 2024; 12(6):1369.

https://doi.org/10.3390/biomedicines12061369

Trigo CM, Rodrigues JS, Camões SP, Solá S, Miranda JP. Mesenchymal stem cell secretome for regenerative medicine: Where do we stand? J Adv Res 2025; 70:103-124

https://doi.org/10.1016/j.jare.2024.05.004

Yang M, Sun W, Xiao L, He M, Gu Y, Yang T, et al. Mesenchymal Stromal Cells Suppress Hippocampal Neuron Autophagy Stress Induced by Hypoxic-Ischemic Brain Damage: The Possible Role of Endogenous IL-6 Secretion. Neural Plast. 2020; 2020:8822579.

https://doi.org/10.1155/2020/8822579

Yari H, Mikhailova MV, Mardasi M, Jafarzadehgharehziaaddin M, Shahrokh S, Thangavelu L, et al. Emerging role of mesenchymal stromal cells (MSCs)-derived exosome in neurodegeneration-associated conditions: a groundbreaking cell-free approach. Stem Cell Res Ther 2022; 13(1):423.

https://doi.org/10.1186/s13287-022-03122-5

Wang F, Chen X, Li J, Wang D, Huang H, Li X, et al. Dose-and time-dependent effects of human mesenchymal stromal cell infusion on cardiac allograft rejection in mice. Stem Cells and Development. 2021 Feb 15;30(4):203-13. https://doi.org/10.1089/scd.2019.0300

https://doi.org/10.1089/scd.2019.0300

Zhang Y, Huang Z, Hao H, Yan M. Hypoxic culture of umbilical cord mesenchymal stem cell-derived sEVs prompts peripheral nerve injury repair. Front Cell Neurosci 2023; 16:897224.

https://doi.org/10.3389/fncel.2022.897224

Wugeng S, Zan S, Liu Y, Bai Y, Hu A, Wei X, et al. MicroRNA-214-3p protects against myocardial ischemia-reperfusion injury by targeting demethylase lysine demethylase 3A. Regenerative Therapy. 2023 Jun 1;23:17-24.

https://doi.org/10.1016/j.reth.2023.01.008

Vohra M, Arora SK. Mesenchymal stem cells: the master immunomodulators. Exploration of Immunology 2023; 3(2):104-22.

https://doi.org/10.37349/ei.2023.00092

Wobma HM, Kanai M, Ma SP, Shih Y, Li HW, Duran-Struuck R, et al. Dual IFN-γ/hypoxia priming enhances immunosuppression of mesenchymal stromal cells through regulatory proteins and metabolic mechanisms. Journal of immunology and regenerative medicine. 2018 Mar 1;1:45-56.

https://doi.org/10.1016/j.regen.2018.01.001

Deuse T, Stubbendorff M, Tang-Quan K, Phillips N, Kay MA, Eiermann T, et al. Immunogenicity and immunomodulatory properties of umbilical cord lining MSCs. Cell Transplant 2011; 20(5):655-67.

https://doi.org/10.3727/096368910X536473

McGettrick AF, O'Neill LAJ. The Role of HIF in Immunity and Inflammation. Cell Metab 2020; 32(4):524-36.

https://doi.org/10.1016/j.cmet.2020.08.002

Huang Y, Wu Q, Tam PK. Immunomodulatory mechanisms of mesenchymal stem cells and their potential clinical applications. International Journal of Molecular Sciences. 2022 Sep 2;23(17):10023.

https://doi.org/10.3390/ijms231710023

Yang M, Sun W, Xiao L, He M, Gu Y, Yang T, et al. Mesenchymal Stromal Cells Suppress Hippocampal Neuron Autophagy Stress Induced by Hypoxic‐Ischemic Brain Damage: The Possible Role of Endogenous IL‐6 Secretion. Neural Plasticity. 2020;2020(1):8822579.

https://doi.org/10.1155/2020/8822579

Published
2025-12-08
How to Cite
1.
Fahreza R, Setiawan E, Trisnadi S, Putra A, Hidayah N, Fikriya Novita Sari, Ibrahim S. Modulation of inflammatory pathways in ischemic stroke rats by hypoxia-primed umbilical cord mesenchymal stem cells (UC-MSCs): Implications for IFN-γ and IL-10 signaling. InaJBCS [Internet]. 2025Dec.8 [cited 2025Dec.21];57(4). Available from: https://journal.ugm.ac.id/v3/InaJBCS/article/view/24529
Section
Articles