Background and Purpose Intestinal barrier dysfunction caused by mitochondrial stress, oxidative damage and apoptosis, are hallmarks of dysbiosis-associated gastrointestinal (GI) disorders. Staurosporine causes downstream features of dysbiosis-induced epithelial damage. KATP (Kir6.x) channels act as metabolic sensors linking cellular energetics to stress adaptation. We investigated whether diazoxide preserved epithelial integrity under staurosporine-induced stress.Experimental Approach HCEC-1CT cells were exposed to staurosporine or in combination with diazoxide. Mitochondrial function, ROS generation, transepithelial electrical resistance (TEER), paracellular permeability, tight junction expression, KATP channel subunit regulation, MAPK signalling and epigenetic markers were assessed using functional assays, RT-qPCR and western blotting. KATP channel involvement was validated using the blocker glibenclamide.Key Results Diazoxide restored mitochondrial function following staurosporine-induced damage in the co-treatment model, with an EC50 of 6 & times; 10-5 M. Reducing ROS accumulation and preserving epithelial barrier integrity, as demonstrated by maintenance of TEER and paracellular sealing. Diazoxide enhanced SUR1/SUR2 expression and glycosylation, preserved ERK1/2 phosphorylation and sustained histone H3 lysine 27 acetylation, supporting active gene transcription under stress. Co-administration with staurosporine prevented downregulation of KATP channel subunits. Diazoxide also attenuated staurosporine-induced dysregulation of pro-inflammatory and apoptotic genes, reinforcing its function as a molecular stabilizer under cytotoxic stress. These protective effects were reversed by glibenclamide, indicating KATP channel dependence.Conclusions and Implications Diazoxide may be a potential therapeutic candidate for oxidative stress-driven barrier dysfunction in gastrointestinal diseases. Future studies incorporating physiologically relevant stressors, such as LPS or H2S, will be important to validate these protective mechanisms in contexts more directly related to microbial dysbiosis.
Modulation of KATP channels by diazoxide preserves mitochondrial function and barrier integrity under staurosporine-induced epithelial stress
Antonacci M.;Di Turi A.;Tricarico D.;
2026-01-01
Abstract
Background and Purpose Intestinal barrier dysfunction caused by mitochondrial stress, oxidative damage and apoptosis, are hallmarks of dysbiosis-associated gastrointestinal (GI) disorders. Staurosporine causes downstream features of dysbiosis-induced epithelial damage. KATP (Kir6.x) channels act as metabolic sensors linking cellular energetics to stress adaptation. We investigated whether diazoxide preserved epithelial integrity under staurosporine-induced stress.Experimental Approach HCEC-1CT cells were exposed to staurosporine or in combination with diazoxide. Mitochondrial function, ROS generation, transepithelial electrical resistance (TEER), paracellular permeability, tight junction expression, KATP channel subunit regulation, MAPK signalling and epigenetic markers were assessed using functional assays, RT-qPCR and western blotting. KATP channel involvement was validated using the blocker glibenclamide.Key Results Diazoxide restored mitochondrial function following staurosporine-induced damage in the co-treatment model, with an EC50 of 6 & times; 10-5 M. Reducing ROS accumulation and preserving epithelial barrier integrity, as demonstrated by maintenance of TEER and paracellular sealing. Diazoxide enhanced SUR1/SUR2 expression and glycosylation, preserved ERK1/2 phosphorylation and sustained histone H3 lysine 27 acetylation, supporting active gene transcription under stress. Co-administration with staurosporine prevented downregulation of KATP channel subunits. Diazoxide also attenuated staurosporine-induced dysregulation of pro-inflammatory and apoptotic genes, reinforcing its function as a molecular stabilizer under cytotoxic stress. These protective effects were reversed by glibenclamide, indicating KATP channel dependence.Conclusions and Implications Diazoxide may be a potential therapeutic candidate for oxidative stress-driven barrier dysfunction in gastrointestinal diseases. Future studies incorporating physiologically relevant stressors, such as LPS or H2S, will be important to validate these protective mechanisms in contexts more directly related to microbial dysbiosis.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
