Intermittent High Glucose Enhances Mitochondria Oxidative Stress In HIT-T15 Cells In Culture

Objective:To evaluate the effects of glucose fluctuation on insulin secretion, mitochondrial membrane potential,ATP contents,malonaldehyde (MDA) level and reactive oxygen species(ROS) level in HIT-T15 cells and to explore the possibly injury mechanism of oxidative stress on glucose fluctuation in β cell line.Subject and Methods: HIT-T15 cells(pancreatic β cell line) were incubated in medium containing persist high glucose (16.7Mm) or intermittent high glucose (5.5-16.7 mM) for at least 72h with or without 1mmol/L N-acetyl-L-cysteine (NAC). Intracellular ROS was evaluated by the staining fluorescent probe of 2',7'- dichlorofluorescein diacetate and then analyzed by flow cytometry. MDA was measured through thiobarbituric acid reaction (TBA). The mitochondrial membrane potential (MMP) was measured by monitoring rhodamine-123 fluorescence intensity using laser scanning confocal microscope. The analysis of cell ATP contents was performed by using the luciferin-luciferase kit. Basal (2.8mmol/L glucose) and stimulated...