Background: Ulcerative colitis (UC) is a typical inflammatory bowel disease requiring long-term management. Although fecal calprotectin (FC) is widely employed for assessing disease activity, it is still considered insufficient as a standalone tool. New biomarkers are needed to better predict risk and comprehensively reflect biological pathways. This study aimed to identify potential fecal biomarkers to monitor disease activity in UC.
Methods: C57BL/6J mice were exposed to dextran sulfate sodium (DSS) treatment for 7 days. Feces were collected and subjected to proteomic analysis and enzyme-linked immunosorbent assay (ELISA). Mouse colon tissues were subjected to histopathological and immunofluorescence analyses. The correlations between the selected fecal proteins and disease severity were evaluated and compared with FC.
Results: Proteomic analysis revealed increases in fecal complement component 3 (C3) and fibronectin (FN) in the DSS group. Next, we measured fecal C3 and FN levels in mice using ELISA. Significant elevation in C3 and FN levels was observed as early as day 1 after DSS treatment, preceding the increase in FC. Both fecal C3 and FN demonstrated significant correlations with disease activity, with C3 exhibiting a stronger correlation than FC. Using immunofluorescence, we observed distinct C3 and FN expressions in both the colonic tissues and the intestinal lumen.
Conclusion: These findings demonstrate that fecal C3 and FN are promising candidate biomarkers for monitoring UC disease activity, and their utility requires further validation in other colitis models and human cohorts.
| [1] |
Rubin DT, Ananthakrishnan AN, Siegel CA, Barnes EL, Long MD. ACG clinical guideline update: ulcerative colitis in adults. Am J Gastroenterol. 2025; 120(6): 1187-1224.
|
| [2] |
Rubin DT, Ananthakrishnan AN, Siegel CA, Sauer BG, Long MD. ACG clinical guideline: ulcerative colitis in adults. Am J Gastroenterol. 2019; 114(3): 384-413.
|
| [3] |
Pabla BS, Schwartz DA. Assessing severity of disease in patients with ulcerative colitis. Gastroenterol Clin N Am. 2020; 49(4): 671-688.
|
| [4] |
Ribeiro da Silva RAV, Genaro LM, Lahan-Martins D, Leal RF, Ayrizono MLS. Comparison between fecal calprotectin and magnetic resonance enterography in ileal Crohn's disease for activity disease assessment: a systematic review. Transl Gastroenterol Hepatol. 2025; 10:73.
|
| [5] |
Caron B, Sequier L, Dignass A, et al. Management of patients with ulcerative proctitis: a global survey. Dig Dis Sci. 2025; 71: 1416-1424.
|
| [6] |
Murray J, Kok KB, Ayling RM. Fecal calprotectin in gastrointestinal disease. Clin Chem. 2023; 69(7): 699-710.
|
| [7] |
Selvakumar B, Samsudin R. Intestinal barrier dysfunction in inflammatory bowel disease: pathophysiology to precision therapeutics. Inflamm Bowel Dis. 2025; 31: 3450-3464.
|
| [8] |
Lee SH, Lopes E, Colombel JF, Ungaro R. Identifying potential targets for the interception of inflammatory bowel disease: toward precision prevention. Inflamm Bowel Dis. 2025; 31(Supplement_2): S51-S60.
|
| [9] |
Alsoud D, Vermeire S, Verstockt B. Biomarker discovery for personalized therapy selection in inflammatory bowel diseases: challenges and promises. Cur Res Pharmacol Drug Dis. 2022; 3:100089.
|
| [10] |
Harder BJ, Lekkerkerker AN, Casavant EP, et al. Comprehensive profiling of the human fecal proteome from IBD patients with DIA-MS enables evaluation of disease-relevant proteins. Proteomics Clin Appl. 2024; 18(5):e2300075.
|
| [11] |
Nowak JK, Kalla R, Satsangi J. Current and emerging biomarkers for ulcerative colitis. Expert Rev Mol Diagn. 2023; 23(12): 1107-1119.
|
| [12] |
Chassaing B, Srinivasan G, Delgado MA, Young AN, Gewirtz AT, Vijay-Kumar M. Fecal lipocalin 2, a sensitive and broadly dynamic non-invasive biomarker for intestinal inflammation. PLoS One. 2012; 7(9):e44328.
|
| [13] |
Zollner A, Schmiderer A, Reider SJ, et al. Faecal biomarkers in inflammatory bowel diseases: calprotectin versus lipocalin-2-a comparative study. J Crohns Colitis. 2021; 15(1): 43-54.
|
| [14] |
Kemper C, Ferreira VP, Paz JT, Holers VM, Lionakis MS, Alexander JJ. Complement: the road less traveled. J Immunol (Baltimore, Md: 1950). 2023; 210(2): 119-125.
|
| [15] |
Tian X, Zhang L, Qian X, et al. Gut complement system: a new frontier in microbiota-host communication and intestinal homeostasis. J Clin Invest. 2025; 135(19):e188349.
|
| [16] |
Wu M, Zheng W, Song X, et al. Gut complement induced by the microbiota combats pathogens and spares commensals. Cell. 2024; 187(4): 897-913.e18.
|
| [17] |
Patten J, Wang K. Fibronectin in development and wound healing. Adv Drug Deliv Rev. 2021; 170: 353-368.
|
| [18] |
Qin Z, Wang R, Zhang Y. Dual roles of complement in ulcerative colitis: insights from clinical studies and animal research. Eur J Med Res. 2025; 30(1):1234.
|
| [19] |
Kolachala VL, Bajaj R, Wang L, et al. Epithelial-derived fibronectin expression, signaling, and function in intestinal inflammation. J Biol Chem. 2007; 282(45): 32965-32973.
|
| [20] |
Rappold R, Kalogeropoulos K, La Regina G, Auf dem Keller U, Slack E, Vogel V. Relaxation of mucosal fibronectin fibers in late gut inflammation following neutrophil infiltration in mice. NPJ Biol Phys Mech. 2025; 2(1):4.
|
| [21] |
Hansson GC. Mucus and mucins in diseases of the intestinal and respiratory tracts. J Intern Med. 2019; 285(5): 479-490.
|
| [22] |
Shin JH, Bozadjieva-Kramer N, Seeley RJ. Reg3γ: current understanding and future therapeutic opportunities in metabolic disease. Exp Mol Med. 2023; 55(8): 1672-1677.
|
| [23] |
Zarantonello A, Revel M, Grunenwald A, Roumenina LT. C3-dependent effector functions of complement. Immunol Rev. 2023; 313(1): 120-138.
|
| [24] |
Peng Q, Li K, Sacks SH, Zhou W. The role of anaphylatoxins C3a and C5a in regulating innate and adaptive immune responses. Inflamm Allergy Drug Targets. 2009; 8(3): 236-246.
|
| [25] |
Sacks SH. Complement fragments C3a and C5a: the salt and pepper of the immune response. Eur J Immunol. 2010; 40(3): 668-670.
|
| [26] |
Biancheri P, Di Sabatino A, Corazza GR, MacDonald TT. Proteases and the gut barrier. Cell Tissue Res. 2013; 351(2): 269-280.
|
| [27] |
Koivisto L, Bi J, Häkkinen L, Larjava H. Integrin αvβ6: structure, function and role in health and disease. Int J Biochem Cell Biol. 2018; 99: 186-196.
|
| [28] |
Sutherland TE, Dyer DP, Allen JE. The extracellular matrix and the immune system: a mutually dependent relationship. Science. 2023; 379(6633):eabp8964.
|
| [29] |
Tshikudi DM, Bernstein CN, Mishra S, Ghia JE, Armstrong HK. Influence of biological sex in inflammatory bowel diseases. Nat Rev Gastroenterol Hepatol. 2025; 22(6): 415-437.
|
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2026 The Author(s). Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.