Abstract:Objective: To investigate the effects of cyanidin-3-O-glucoside (C3G) on islet beta cell injury and its mechanism in high glucose and high fat (HGHF) environment. Methods: The pancreatic β cells were treated with C3G (10, 20, 30, 40, 50 μmol/L) for 24 h, and the cell viability was detected by CCK-8 assay. The HGHF environment was established by using the culture medium containing 25 mmol/L glucose and 0.5 mmol/L palmitate, and C3G (10, 20, 30, 40, 50 μmol/L) was added to treat the pancreatic β cells for 24 h. The cell viability was detected by CCK-8 assay. The experiment was divided into four groups: control group, HGHF group, HGHF+C3G group, and HGHF+C3G+3-MA group. After the corresponding treatment, CCK-8 assay was used to detect the cell viability of each group. DCFH-DA fluorescent probe was used to detect the ROS level of each group. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) in each group were detected by colorimetric assay. The insulin secretion of each group was determined by ELISA assay. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the mRNA expression levels of autophagy-related genes 5 (Atg5), microtubule-associated protein light chain 3 (LC3), and Beclin-1 in each group. Western blot was used to detect the protein expression levels of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), and activated caspase-3 (Cleaved Caspase-3) in each group. Results: Compared with the control group, the cell viability of the C3G group (10, 20, 30, 40, 50 μmol/L) was increased (P<0.05), and the cell viability of the HGHF group was decreased (P<0.05). Compared with the HGHF group, the addition of 20, 30, 40, 50 μmol/L C3G in HGHF environment could significantly increase the cell viability (P<0.05). Compared with the HGHF group, the cell viability of the HGHF+C3G group was increased (P<0.05), the ROS content and MDA content were decreased (P<0.05), the SOD activity and GSH-Px content were increased (P<0.05), the insulin secretion level was increased (P<0.05), the mRNA relative expression levels of Atg5, LC3, and Beclin-1 were up-regulated (P<0.05), the LC3-Ⅱ/LC3-Ⅰ ratio and Beclin-1 protein relative expression levels were up-regulated (P<0.05), the Bax and Cleaved Caspase-3 protein relative expression levels were down-regulated, and Bcl-2 protein relative expression level was up-regulated (P<0.05). However, when C3G was added to the HGHF environment and 3-methyladenine (3-MA), an autophagy inhibitor, was also used, the protective effect of C3G on pancreatic β cells in HGHF environment disappeared. Conclusion: C3G has a protective effect on pancreatic β cells in HGHF environment. It can improve cell viability, inhibit oxidative stress damage, and this effect may be related to the promotion of autophagy.
李恒, 张翠兰. 高糖高脂环境下抑制自噬对矢车菊素-3-O-葡萄糖苷保护胰岛β细胞损伤的影响[J]. 河北医学, 2024, 30(2): 182-188.
LI Heng, ZHANG Cuilan. Effect of Inhibition of Autophagy on Protection of β-Cell Damage by Cyanidin-3-O-Glucoside in High Glucose and High Fat Environment. HeBei Med, 2024, 30(2): 182-188.
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