A Solini1, G Sebastiani2, L Nigi2, E Santini3, C Rossi3, F Dotta4. 1. Department of surgical, medical, molecular and critical area pathology, university of Pisa, Via Roma 67, 56126 Pisa, Italy. Electronic address: anna.solini@med.unipi.it. 2. Department of medicine, surgery and neuroscience, university of Siena and Fondazione Umberto di Mario-Toscana life science, Viale Bracci 18, 53100 Siena, Italy. 3. Department of clinical and experimental medicine, university of Pisa, Pisa, Italy. 4. Department of medicine, surgery and neuroscience, university of Siena and Fondazione Umberto di Mario-Toscana life science, Viale Bracci 18, 53100 Siena, Italy. Electronic address: francesco.dotta@unisi.it.
Abstract
AIM: SGLT2 inhibitors reduce renal glucose uptake through an insulin-independent mechanism. They also increase glucagon concentration, although the extent to which this is due to a direct effect on pancreatic alpha cells remains unclear. METHODS: In the present work, αTC1 cells treated with the SGLT2 inhibitor dapagliflozin (Dapa) were analyzed for glucose transporters, molecular mediators of hormone secretion, glucagon and GLP-1 release, and the effects of somatostatin. Data were validated in murine and human pancreatic islets. RESULTS: SLC5A2 (the SGLT2-encoding gene) was nearly undetectable in αTC1 cells, not even by a digital PCR technique using different probes. In contrast, SLC5A1 (the SGLT1-encoding gene) was constitutively abundant in αTC1 cells and in islets, and increased with Dapa. This was associated with greater glucagon release, preceded by increased expression of preproglucagon and HNF4α. Looking at the candidate intracellular signalling pathway, reduced PASK and increased AMPK-α2 expression were also detected. GLUT1 and GLUT2, as well as regulators of glucagon release and alpha-cell phenotype (chromogranin A, paired box 6, proprotein convertase 1/2, synaptophysin), were unaffected by Dapa, as were GLP-1 receptor expression and GLP-1 release. Low glucose did not influence the stimulatory effect of Dapa on glucagon release, but was instead almost fully reverted by SLC5A1 silencing. When the effect of Dapa on AMPK and PASK, emerging regulators of lipid and glucose metabolism, was tested, upregulated AMPK-α2 appeared to be involved in molecular signalling. CONCLUSION: Our study has shown that, in αTC1 cells, Dapa acutely upregulates SGLT1 expression and increases glucagon release through an SGLT1-dependent mechanism, with SGLT2 expression virtually undetectable. These results suggest the involvement of SGLT1 in modulating glucagon increases following SGLT2 inhibition.
AIM: SGLT2 inhibitors reduce renal glucose uptake through an insulin-independent mechanism. They also increase glucagon concentration, although the extent to which this is due to a direct effect on pancreatic alpha cells remains unclear. METHODS: In the present work, αTC1 cells treated with the SGLT2 inhibitor dapagliflozin (Dapa) were analyzed for glucose transporters, molecular mediators of hormone secretion, glucagon and GLP-1 release, and the effects of somatostatin. Data were validated in murine and humanpancreatic islets. RESULTS:SLC5A2 (the SGLT2-encoding gene) was nearly undetectable in αTC1 cells, not even by a digital PCR technique using different probes. In contrast, SLC5A1 (the SGLT1-encoding gene) was constitutively abundant in αTC1 cells and in islets, and increased with Dapa. This was associated with greater glucagon release, preceded by increased expression of preproglucagon and HNF4α. Looking at the candidate intracellular signalling pathway, reduced PASK and increased AMPK-α2 expression were also detected. GLUT1 and GLUT2, as well as regulators of glucagon release and alpha-cell phenotype (chromogranin A, paired box 6, proprotein convertase 1/2, synaptophysin), were unaffected by Dapa, as were GLP-1 receptor expression and GLP-1 release. Low glucose did not influence the stimulatory effect of Dapa on glucagon release, but was instead almost fully reverted by SLC5A1 silencing. When the effect of Dapa on AMPK and PASK, emerging regulators of lipid and glucose metabolism, was tested, upregulated AMPK-α2 appeared to be involved in molecular signalling. CONCLUSION: Our study has shown that, in αTC1 cells, Dapa acutely upregulates SGLT1 expression and increases glucagon release through an SGLT1-dependent mechanism, with SGLT2 expression virtually undetectable. These results suggest the involvement of SGLT1 in modulating glucagon increases following SGLT2 inhibition.
Authors: Corey B Hughes; George M Mussman; Phil Ray; Robert C Bunn; Virgilius Cornea; Kathryn M Thrailkill; John L Fowlkes; Iuliana Popescu Journal: Cell Tissue Res Date: 2021-01-06 Impact factor: 5.249
Authors: Rune E Kuhre; Seyed M Ghiasi; Alice E Adriaenssens; Nicolai J Wewer Albrechtsen; Daniel B Andersen; Alexander Aivazidis; Lihua Chen; Thomas Mandrup-Poulsen; Cathrine Ørskov; Fiona M Gribble; Frank Reimann; Nils Wierup; Björn Tyrberg; Jens J Holst Journal: Diabetologia Date: 2019-03-22 Impact factor: 10.122