Effect of artificial sweeteners on the insulin signaling pathway of human neuroblastoma cells SH-SY5Y

Abstract

Uvod: Neurodegenerativne bolesti predstavljaju sve veći zdravstveni problem u populacijama zemalja razvijenog svijeta. Nekoliko istraživanja ukazuje na povezanost inzulinske rezistencije s razvojem Alzheimerove i Parkinsonove bolesti. Sukraloza podiže razinu glukoze i inzulina kod pretilih ženama. Vrlo male doze natrijeva ciklamata mogu poremetiti diferencijaciju i proliferaciju osteoblasta. Ciljevi: Ciljevi ovog istraživanja su bili ispitati učinke tretmana triju koncentracija natrijeva ciklamata (1 mM, 10 mM, 100 mM) i sukraloze (0,2 mM, 2 mM, 20 mM) samostalno, te u kombinacijama s inzulinom i levodopom na preživljenje, inzulinski signalni put i metabolički odgovor diferenciranih stanica ljudskog neuroblastoma. Materijali i metode: Diferencirane SH-SY5Y stanice su tretirane sladilima i dodatnim tretmanima. Citotoksičan učinak tretmana određen je MTT testom. Aktivnost inzulinskog signalnog puta ispitala se Western blot metodom. Ukupni učinak tretmana na metabolički odgovor tretiranih stanica ispitan je MALDI-TOF MS metodom. Rezultati: Stanice tretirane sukralozom u sve tri koncentracije gdje je kao dodatni tretman korištena i levodopa imale su značajno veće preživljenje u odnosu na netretiranu ili inzulinom tretiranu kontrolnu skupinu. Natrijev ciklamat je na preživljenje stanica imao o koncentraciji ovisan učinak. Najmanja koncentracija značajno je povećala preživljenje stanice neovisno o dodatnom tretmanu inzulinom ili levodopom. Zaključak: Sukraloza i natrijev ciklamat u kombinaciji s inzulinom i levodopom sprječavaju apoptozu diferenciranih stanica ljudskog neuroblastoma u određenim koncentracijama putem inzulinskog signalnog puta. Međutim, potrebno je istražiti učinak ovih sladila na ostale metaboličke puteve.Introduction: Neurodegenerative diseases represent an increasing health problem in the populations of countries in the Western world. Several studies indicate a connection between insulin resistance and the development of Alzheimer's and Parkinson's diseases. Sucralose raises glucose and insulin levels in obese women. Very small doses of sodium cyclamate can disrupt the differentiation and proliferation of osteoblasts. Objectives: The objectives of this research were to examine the effects of three concentrations of sodium cyclamate (1 mM, 10 mM, 100 mM) and sucralose (0,2 mM, 2 mM, 20 mM) independently and in combination with insulin and levodopa on survival, insulin signaling pathway and metabolic response of differentiated human neuroblastoma cells. Material and methods: Differentiated SH-SY5Y cells were treated with sweeteners and additional treatments. The cytotoxic effect of the treatment was determined by the MTT test. The activity of the insulin signaling pathway was examined using the Western blot method. The overall effect on the metabolic response of the treated cells was determined by the MALDI-TOF MS method. Results: Cells treated with sucralose in all three concentrations, where levodopa was also used as an additional treatment, had significantly higher survival compared to the untreated or insulin-treated control group. Sodium cyclamate had a concentration-dependent effect on SH-SY5Y cells survival. The lowest concentration significantly increased cell survival independent of additional treatment with insulin or levodopa. Conclusion: Sucralose and sodium cyclamate in combination with insulin and levodopa prevent apoptosis of differentiated human neuroblastoma cells in certain concentrations via the insulin signaling pathway. However, it is necessary to investigate the effect of these sweeteners on other metabolic pathways