Abstract
Membrane-bound guanylyl cyclase (GC-G), a noncanonical receptor, has been implicated in intestinal epithelial homeostasis. This study investigated the role of GC-G in intestinal ischemia-reperfusion (IIR) injury and its involvement in nitric oxide (NO)–related responses using male wild-type (WT) and GC-G knockout (KO) mice. Mice underwent superior mesenteric artery occlusion followed by 3- or 24-hour reperfusion and were intraperitoneally administered arginine (NO donor), aminoguanidine (selective iNOS inhibitor), or L-NAME (non-selective NOS inhibitor). In WT mice, IIR reduced jejunal mass, increased IL-6 and TUNEL-staining at 3 h, and suppressed nitrate/nitrite (NOx) and Ki-67 at 24 h. Arginine reduced caspase-3 activity at 3 h and enhanced Ki-67 and DNA content at 24 h. Aminoguanidine suppressed early apoptosis and improved later proliferation. L-NAME reduced early injury but enhanced later proliferation without restoring DNA content. GC-G deficiency was associated with attenuated early apoptotic responses but impaired mucosal regeneration following IIR. In KO mice, arginine failed to enhance proliferation and was associated with elevated apoptosis and cytokines, while NOS inhibitors reduced systemic NOx and IL-6 without improving epithelial proliferation or altering caspase-3 activity or bulk cGMP levels. Notably, intestinal NOx levels remained elevated in KO mice. Together, these findings suggest that GC-G contributes to the coordination of NO-related epithelial responses and influences intestinal sensitivity to NO-associated interventions during IIR, without detectable changes in total tissue cGMP levels.
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Acknowledgements
We thank Ching-Yi Chen, Mei-Ling Li, Chun-Hsiang Cheng, and Hui-Ru Tsai for their technical support allowing for the completion of this study.
Funding
This work is supported by the National Science Council of the Republic of China under the grant number NSC 96-2320-B-309-003-MY2 and by Changhua Christian Hospital under the grant number 102-CCH-IRP-025. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Conceptualization, H.-C.L. and R.-B.Y.; methodology, H.-C.L. and C.-H.L.; validation, H.-C.L. and R.-B.Y.; formal analysis, H.-C.L. and R.-B.Y.; investigation, C.-H.L.; data curation, R.-B.Y., H.-C.L. and C.-H.L.; writing—original draft preparation, H.-C.L.; writing—review and editing, R.-B.Y. and C.-H.L. supervision, H.-C.L. and C.-H.L.; project administration, H.-C.L. and R.-B.Y.; funding acquisition, H.-C.L. All authors read and approved the final manuscript.
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41598_2026_47089_MOESM1_ESM.jpg (download JPG )
Supplementary Material 1Representative hematoxylin and eosin (H&E)–stained light micrographs of jejunal sections from WT and KO mice subjected to sham operation or intestinal ischemia followed by 3-hour reperfusion (A) or 24-hour (B) reperfusion. Scale bar = 100 μm.
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Supplementary Material 2Experimental design of IIR groups. WT or KO mice underwent sham operation (WTS, KOS) or 45 min intestinal ischemia followed by 3 or 24 h reperfusion. WT mice received saline (C3W, C24W), arginine (300 mg/kg; A3W, A24W), aminoguanidine (100 mg/kg; G3W, G24W), or L-NAME (10 mg/kg; N3W, N24W), while KO mice underwent parallel treatments (C3K/C24K, A3K/A24K, G3K/G24K, N3K/N24K).
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Supplementary Material 3 Summary of three-way ANOVA results (genotype × reperfusion time × treatment) for key outcomes following intestinal ischemia–reperfusion.
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Lo, HC., Yang, RB. & Lee, CH. Membrane-bound guanylyl cyclase-G deficiency impairs nitric oxide–mediated epithelial repair in intestinal ischemia-reperfusion injury in male mice. Sci Rep (2026). https://doi.org/10.1038/s41598-026-47089-2
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DOI: https://doi.org/10.1038/s41598-026-47089-2
