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In Vivo and In Vitro Analysis of MAPK and PI3K/AKT Signaling Dysregulation in Colonic Epithelial Injury

Martín Alejandro Fernández,PhD1, Lucía Valentina Rojas, MD, PhD  ORCID2*

1 Department of Gastroenterology and Experimental Pathophysiology, Faculty of Medical Sciences, National University of La Plata
La Plata, Buenos Aires, Argentina.
2 Institute of Biomedical Research (BIOMED), Pontifical Catholic University of Argentina, Buenos Aires, Argentina.

DOI: 10.18081/2378-5225/14.75  
Cited by 0

 Article history: Received 30 December 2024 · Revised 21 February 2025 · Accepted 12 March 2025 · Published 05 April 2025

© 2025 Rojas, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (CC BY 4.0). 

CC BY 4.0                                                                                                   

Abstract

Background: Colonic epithelial injury is a central feature of many gastrointestinal disorders and is driven by complex interactions between inflammatory signaling, oxidative stress, and dysregulated cell survival pathways. Among these, the mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathways play critical but often opposing roles in regulating epithelial cell fate. However, the integrated contribution of these pathways to colonic injury remains incompletely understood.

Objective: This study aimed to investigate the dysregulation of MAPK and PI3K/AKT signaling pathways in colonic epithelial injury using complementary in vivo and in vitro experimental models.

Methods: Colonic injury was induced in mice using dextran sulfate sodium (DSS) to establish an in vivo model of epithelial damage. Histopathological evaluation, oxidative stress markers, cytokine levels, and signaling pathway activation were assessed. In parallel, human colonic epithelial cell lines (Caco-2 and HT-29) were exposed to oxidative (H₂O₂) and inflammatory (TNF-α) stress to model epithelial injury in vitro. Cell viability, apoptosis, reactive oxygen species (ROS) production, barrier function, and signaling pathway modulation were analyzed. Pharmacologic inhibitors were used to evaluate the specific roles of MAPK and PI3K/AKT pathways.

Results: DSS-induced colitis resulted in significant epithelial damage, increased oxidative stress, elevated pro-inflammatory cytokines, and activation of MAPK signaling pathways (ERK, JNK, and p38). In contrast, PI3K/AKT signaling showed reduced activity in severe injury states. MAPK inhibition attenuated inflammation, oxidative stress, apoptosis, and barrier disruption, while PI3K/AKT inhibition exacerbated epithelial injury and cell death. In vitro findings confirmed that oxidative and inflammatory stress decreased cell viability, increased ROS production, and induced apoptosis, effects that were mitigated by MAPK inhibition but worsened by suppression of PI3K/AKT signaling. Barrier integrity assays demonstrated that PI3K/AKT signaling is essential for maintaining tight junction function, whereas MAPK activation contributes to barrier breakdown.

Conclusions: These findings demonstrate that colonic epithelial injury is driven by an imbalance between MAPK and PI3K/AKT signaling pathways. MAPK overactivation promotes inflammatory and apoptotic responses, while PI3K/AKT signaling provides critical cytoprotective and barrier-preserving functions. Targeted modulation of these pathways may represent a promising therapeutic strategy for gastrointestinal diseases characterized by epithelial injury.

Keywords: Colonic epithelial injury; MAPK signaling pathway; PI3K/AKT pathway; Oxidative stress; Reactive oxygen species (ROS

References

  1. Xavier RJ, Podolsky DK. Unravelling the pathogenesis of inflammatory bowel disease. Nature. 2007;448(7152):427–434. doi:10.1038/nature06005
  2. Turner JR. Intestinal mucosal barrier function in health and disease. Nat Rev Immunol. 2009;9(11):799–809. doi:10.1038/nri2653
  3. Karin M. The regulation of AP-1 activity by mitogen-activated protein kinases. J Biol Chem. 1995;270(28):16483–16486. doi:10.1074/jbc.270.28.16483
  4. Pearson G, Robinson F, Beers Gibson T, et al. Mitogen-activated protein kinase pathways: regulation and physiological functions. Endocr Rev. 2001;22(2):153–183. doi:10.1210/edrv.22.2.0428
  5. Johnson GL, Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science. 2002;298(5600):1911–1912. doi:10.1126/science.1072682
  6. Cantley LC. The phosphoinositide 3-kinase pathway. Science. 2002;296(5573):1655–1657. doi:10.1126/science.296.5573.1655
  7. Manning BD, Cantley LC. AKT/PKB signaling: navigating downstream. Cell. 2007;129(7):1261–1274. doi:10.1016/j.cell.2007.06.009
  8. Bhattacharyya S, Dudeja PK, Tobacman JK. Lipopolysaccharide activates NF-κB by TLR4 signaling in human intestinal epithelial cells. Am J Physiol Gastrointest Liver Physiol. 2008;295(4):G703–G711. doi:10.1152/ajpgi.90224.2008
  9. Neurath MF. Cytokines in inflammatory bowel disease. Nat Rev Immunol. 2014;14(5):329–342. doi:10.1038/nri3661
  10. Kamada N, Seo SU, Chen GY, Núñez G. Role of the gut microbiota in immunity and inflammatory disease. Nat Rev Immunol. 2013;13(5):321–335. doi:10.1038/nri3430
  11. Grivennikov SI, Greten FR, Karin M. Immunity, inflammation, and cancer. Cell. 2010;140(6):883–899. doi:10.1016/j.cell.2010.01.025
  12. D’Haens G, Sartor RB, Silverberg MS, Petersson J, Rutgeerts P. Future directions in inflammatory bowel disease management. J Crohns Colitis. 2014;8(8):726–734. doi:10.1016/j.crohns.2014.02.025
  13. Wirtz S, Neufert C, Weigmann B, Neurath MF. Chemically induced mouse models of intestinal inflammation. Nat Protoc. 2007;2(3):541–546. doi:10.1038/nprot.2007.41
  14. Hidalgo IJ, Raub TJ, Borchardt RT. Characterization of the human colon carcinoma cell line (Caco-2). Gastroenterology. 1989;96(3):736–749. doi:10.1016/0016-5085(89)90952-5
  15. Reuter S, Gupta SC, Chaturvedi MM, Aggarwal BB. Oxidative stress, inflammation, and cancer. Free Radic Biol Med. 2010;49(11):1603–1616. doi:10.1016/j.freeradbiomed.2010.09.006
  16. Wallace DC. Mitochondria and cancer. Nat Rev Cancer. 2012;12(10):685–698. doi:10.1038/nrc3365
  17. Suzuki T. Regulation of intestinal epithelial permeability by tight junctions. Cell Mol Life Sci. 2013;70(4):631–659. doi:10.1007/s00018-012-1070-x
  18. Terzić J, Grivennikov S, Karin E, Karin M. Inflammation and colon cancer. Gastroenterology. 2010;138(6):2101–2114. doi:10.1053/j.gastro.2010.01.058
  19. Oeckinghaus A, Hayden MS, Ghosh S. Crosstalk in NF-κB signaling pathways. Nat Immunol. 2011;12(8):695–708. doi:10.1038/ni.2065
  20. Luo J, Manning BD, Cantley LC. Targeting the PI3K-AKT pathway in human cancer. Cancer Cell. 2003;4(4):257–262. doi:10.1016/S1535-6108(03)00248-4
  21. Dong C, Davis RJ, Flavell RA. MAP kinases in the immune response. Annu Rev Immunol. 2002;20:55–72. doi:10.1146/annurev.immunol.20.091701.151133
  22. Schulzke JD, Ploeger S, Amasheh M, et al. Epithelial tight junctions in intestinal inflammation. Ann N Y Acad Sci. 2009;1165:294–300. doi:10.1111/j.1749-6632.2009.04062.x
  23. Günzel D, Yu AS. Claudins and the modulation of tight junction permeability. Physiol Rev. 2013;93(2):525–569. doi:10.1152/physrev.00019.2012
  24. Kim EK, Choi EJ. Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta. 2010;1802(4):396–405. doi:10.1016/j.bbadis.2009.12.009
  25. Fresno Vara JA, Casado E, de Castro J, et al. PI3K/Akt signalling pathway and cancer. Cancer Treat Rev. 2004;30(2):193–204. doi:10.1016/j.ctrv.2003.07.007
  26. Banan A, Zhang Y, Fields JZ, Keshavarzian A. PKC and MAPK signaling in ethanol-induced intestinal barrier dysfunction. Am J Physiol Gastrointest Liver Physiol. 2000;279:G621–G633. doi:10.1152/ajpgi.2000.279.3.G621
  27. Al-Sadi R, Boivin M, Ma T. Mechanism of cytokine modulation of epithelial tight junction barrier. Front Biosci. 2009;14:2765–2778. doi:10.2741/3413
  28. Pastorelli L, De Salvo C, Mercado JR, Vecchi M, Pizarro TT. Central role of the gut epithelial barrier. Inflamm Bowel Dis. 2013;19(6):1308–1316. doi:10.1097/MIB.0b013e318281f2a8
  29. Förster C. Tight junctions and the modulation of barrier function. Cell Tissue Res. 2008;334(1):1–9. doi:10.1007/s00441-008-0680-z
  30. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–674. doi:10.1016/j.cell.2011.02.013
  31. Khor B, Gardet A, Xavier RJ. Genetics and pathogenesis of inflammatory bowel disease. Nature. 2011;474(7351):307–317. doi:10.1038/nature10209
  32. Ivanov AI, Parkos CA, Nusrat A. Cytoskeletal regulation of epithelial barrier function. Am J Pathol. 2010;177(2):512–524. doi:10.2353/ajpath.2010.100168
  33. Zha J, Harada H, Yang E, et al. Serine phosphorylation of death agonist BAD. Cell. 1996;87(4):619–628. doi:10.1016/S0092-8674(00)81382-3
  34. Datta SR, Brunet A, Greenberg ME. Cellular survival: a play in three Akts. Genes Dev. 1999;13(22):2905–2927. doi:10.1101/gad.13.22.2905
  35. Cargnello M, Roux PP. Activation and function of the MAPKs. Microbiol Mol Biol Rev. 2011;75(1):50–83. doi:10.1128/MMBR.00031-10
  36. Li Q, Verma IM. NF-κB regulation in the immune system. Nat Rev Immunol. 2002;2(10):725–734. doi:10.1038/nri910
  37. Arthur JS, Ley SC. Mitogen-activated protein kinases in innate immunity. Nat Rev Immunol. 2013;13(9):679–692. doi:10.1038/nri3495
  38. Vivanco I, Sawyers CL. The phosphatidylinositol 3-Kinase–AKT pathway in human cancer. Nat Rev Cancer. 2002;2(7):489–501. doi:10.1038/nrc839
  39. Zeng H, Carlson AQ, Guo Y, et al. Orphan nuclear receptor NR4A1 regulates oxidative stress. Mol Cell Biol. 2014;34(3):485–498. doi:10.1128/MCB.00984-13
  40. Hooper LV, Macpherson AJ. Immune adaptations that maintain homeostasis. Nat Rev Immunol. 2010;10(3):159–169. doi:10.1038/nri2710

BM-Publisher · Pathophysiology of Cell Injury Journal (PCIJ) · E-ISSN 2378-5225 · DOI Prefix 10.18081/pcij/2378-5225

Pathophysiology of Cell Injury Journal (PCIJ)
E-ISSN 2378-5225 · Biannual
BM-Publisher (London, UK)
Open Access

Vol 14, Issue 1 (June 2025), pp. 59–75

How to cite (AMA)

Fernández MA, Rojas LV. In Vivo and In Vitro Analysis of MAPK and PI3K/AKT Signaling Dysregulation in Colonic Epithelial Injury. Pathophysiology of Cell Injury Journal (PCIJ). 2025;14(1):59–75. doi: 10.18081/2378-5225/14.75.

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