A transmural oxidative imbalance has been reported in the different clinical phenotypes of Diverticular Disease (DD) (Pallotta L,2022; Cappelletti M,2022) but the genesis of the oxidative stress remains to be defined. Since the central role of the mitochondria for the metabolism and radicals production, aim of the study is to elucidate the presence of mitochondria alterations in the different DD clinical phenotypes by redox state evaluation, molecular and functional analyses. Methods: Smooth muscle longitudinal and circular cells and tissues were isolated separately from surgical specimens of complicated stenotic DD (cDD, n=5), up-stream non-occlusive colon cancer with diverticula (DIV, n=7) and without diverticula (controls, n=6). Mitochondrial ROS (mtROS), membrane potential (mtMP) and mass (mtM) were evaluated in cells by FACS. Complex II of the mithocondria respiratory electron chain (succinate dehydrogenase, SDH) and mtDNA were evaluated on tissues, respectively by Western Blot and qPCR. Functional cells activity was tested by image micrometry assessing contraction to acetylcholine and relaxation to VIP, before and after 24h/incubation with polyphenols 14mM (Phenolea Active Complex L). Data (mean±SE) were expressed as fold variations from controls, p value<0.05 considered significant (*). Results: In cDD longitudinal muscle, an increase in mtROS (1.4±0.5) was observed, paralleled to an increase in mtMP (2.3*±0.9) and mtM (2.5±1.0), index of radicals overproduction and mitochondrial biogenesis. In DIV, only an increase in mtM (2.3±1.2) was present. Both in DIV and cDD, an oxidation and subsequently loss of SDH was registered (DIV:0.7±0.2; cDD:0.2±0.07), related to radicals overproduction, paralleled to an increase of the ratio nuclear DNA/mtDNA (DIV:1.5±0.2; cDD:1.7±1.1), indicative of mtDNA decrease. In circular muscle, similar oxidative alterations were observed only in cDD, with an increase in mtROS (1.8±0.7), mtMP (3.2±1.3) and mtM (cDD:3.2±1.3) with decrease of SDH (0.2±0.1). Both in DIV and cDD muscle layers, an impairement of functional activity was present. Polyphenols were able to restore LM relaxation (DIV:85.4%±51.1; cDD:62.0%±15.9). Conclusion: A mitochondrial-related oxidative imbalance is present in the different DD phenotypes, suggesting the presence of a primary myopathy. Functional muscular alterations can in part be reverted by polyphenol treatment, providing a possible new tailored-therapeutic and prevention strategies.
T.02.5 OXIDATIVE STRESS IN DIVERTICULAR DISEASE: MITOCHONDRIA INVOLVEMENT OF IN THE DIFFERENT CLINICAL PHENOTYPES / Cappelletti, M.; Vona, R.; Pisano, A.; Gioia, A.; Castagneto Gissey, L.; Tarallo, M.; Matarrese, P.; Giordano, C.; Severi, C.; Pallotta, L.. - In: DIGESTIVE AND LIVER DISEASE. - ISSN 1590-8658. - 55:(2023), pp. 77-209. (Intervento presentato al convegno 29th National Congress of Digestive Disease Italian Federation of Societies of Digestive Diseases – FISMAD tenutosi a Roma) [10.1016/S1590-8658(23)00353-5].
T.02.5 OXIDATIVE STRESS IN DIVERTICULAR DISEASE: MITOCHONDRIA INVOLVEMENT OF IN THE DIFFERENT CLINICAL PHENOTYPES
Cappelletti, M.;Pisano, A.;Castagneto Gissey, L.;Tarallo, M.;Giordano, C.;Severi, C.;Pallotta, L.
2023
Abstract
A transmural oxidative imbalance has been reported in the different clinical phenotypes of Diverticular Disease (DD) (Pallotta L,2022; Cappelletti M,2022) but the genesis of the oxidative stress remains to be defined. Since the central role of the mitochondria for the metabolism and radicals production, aim of the study is to elucidate the presence of mitochondria alterations in the different DD clinical phenotypes by redox state evaluation, molecular and functional analyses. Methods: Smooth muscle longitudinal and circular cells and tissues were isolated separately from surgical specimens of complicated stenotic DD (cDD, n=5), up-stream non-occlusive colon cancer with diverticula (DIV, n=7) and without diverticula (controls, n=6). Mitochondrial ROS (mtROS), membrane potential (mtMP) and mass (mtM) were evaluated in cells by FACS. Complex II of the mithocondria respiratory electron chain (succinate dehydrogenase, SDH) and mtDNA were evaluated on tissues, respectively by Western Blot and qPCR. Functional cells activity was tested by image micrometry assessing contraction to acetylcholine and relaxation to VIP, before and after 24h/incubation with polyphenols 14mM (Phenolea Active Complex L). Data (mean±SE) were expressed as fold variations from controls, p value<0.05 considered significant (*). Results: In cDD longitudinal muscle, an increase in mtROS (1.4±0.5) was observed, paralleled to an increase in mtMP (2.3*±0.9) and mtM (2.5±1.0), index of radicals overproduction and mitochondrial biogenesis. In DIV, only an increase in mtM (2.3±1.2) was present. Both in DIV and cDD, an oxidation and subsequently loss of SDH was registered (DIV:0.7±0.2; cDD:0.2±0.07), related to radicals overproduction, paralleled to an increase of the ratio nuclear DNA/mtDNA (DIV:1.5±0.2; cDD:1.7±1.1), indicative of mtDNA decrease. In circular muscle, similar oxidative alterations were observed only in cDD, with an increase in mtROS (1.8±0.7), mtMP (3.2±1.3) and mtM (cDD:3.2±1.3) with decrease of SDH (0.2±0.1). Both in DIV and cDD muscle layers, an impairement of functional activity was present. Polyphenols were able to restore LM relaxation (DIV:85.4%±51.1; cDD:62.0%±15.9). Conclusion: A mitochondrial-related oxidative imbalance is present in the different DD phenotypes, suggesting the presence of a primary myopathy. Functional muscular alterations can in part be reverted by polyphenol treatment, providing a possible new tailored-therapeutic and prevention strategies.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
