葡萄籽原花青素通过Nrf2信号通路拮抗高糖高脂诱导的MIN6细胞铁死亡

摘要/Abstract

摘要： 目的：铁死亡可能是高糖高脂诱导胰岛β细胞死亡的重要机制，本研究探讨葡萄籽原花青素提取物（grape seed procyanidin extract，GSPE）对高糖高脂诱导胰岛β细胞铁死亡的作用机制。方法：用25 mmol/L 高糖（high glucose）和200 μmol/L棕榈酸钠（sodium palmitate）干预MIN6细胞建立胰岛β细胞损伤模型，进一步用 0、10、20和30 mg/L的GSPE以及Nrf2 siRNA进行干预，用细胞计数试剂盒（The Cell Counting Kit-8 assay, CCK8）检测细胞活性，用试剂盒检测细胞内谷胱甘肽（glutathione, GSH）、丙二醛（malonaldehyde, MDA）、活性氧（reactive oxygen species, ROS）水平，用Western Blot检测核转录因子E2相关因子2（nuclear factor erythroid 2-related factor 2, Nrf2）、血红素氧化酶1（heme oxygenase-1, HO-1）、醌氧化还原酶1（NADPH: quinone oxido-reductase-1，NQO1）、铁代谢指标转铁蛋白受体1（transferrin receptor protein 1, TfR1）、二价金属离子蛋白转运体（divalent metal transporter 1, DMT1）、铁蛋白（ferritin），铁死亡的指标蛋白溶质载体家族7成员11（solute carrier family 7 member 11, SLC7A11）、谷胱甘肽过氧化物酶4（glutathione peroxidase 4, GPX4)、酰基合成酶长链家族成员4（acylcoa synthetase long-chain family member 4, ACSL4）蛋白表达量。结果：与对照组相比，高糖高脂组导致细胞内铁沉积上调TfR1、DMT1、ferritin蛋白表达水平，增加氧化产物MDA、ROS、ACSL4水平，降低GSH水平、下调GPX4、SLC7A11的表达水平而GSPE的干预能够激活Nrf2/HO-1信号通路上调GPX4、SLC7A11蛋白水平，增加GSH水平从而降低MDA、ROS、ACSL4水平和铁代谢指标水平TfR1、DMT1、ferritin来改善高糖高脂诱导的MIN6细胞铁死亡。联合Nrf2 siRNA干预后发现抑制Nrf2后会抑制GSPE的保护作用，抗氧化酶HO-1、GPX4、SLC7A11的表达降低。结论：GSPE可能是通过激活MIN6细胞内的Nrf2信号通路，上调抗氧化酶，减轻高糖高脂诱导的细胞铁死亡，从而提高MIN6细胞存活率。

关键词: 铁死亡, 葡萄籽原花青素, 核转录因子E2相关因子2, MIN6细胞

Abstract: Objective: Ferroptosis may be an important mechanism of high glucose and high fat induced pancreatic β cell death. This study investigated the mechanism of grape seed procyanidin extract (GSPE) on ferroptosis in pancreatic β cells induced by high glucose and high fat. Method: The islet β cell injury model was established by treating MIN6 cells with 25 mmol/L of high glucose and 200 μmol/L of high fat. Then we transfected it with 0, 10, 20 and 30 mg/L GSPE and Nrf2 small interference RNA. The cell activity was detected by the Cell Counting Kit-8 assay (CCK8) kit, and the levels of intracellular glutathione (GSH), malonaldehyde (MDA) and reactive oxygen species (ROS) were detected by kit. The protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NADPH: quinone oxido-reductase-1 (NQO1), iron metabolism indicator transferrin receptor 1 (TfR1), divalent metal ion protein transporter 1 (DMT1), ferritin, and solute carrier family 7 member 11 (SLC7A11) were measured by Western Blot. The protein expression of glutathione peroxidase 4 (GPX4) and acylcoa synthetase long-chain family member 4 (ACSL4) were also measured by Western Blot. Result: Compared with the control group, the high glucose and high fat caused the intracellular iron deposition, up-regulated the protein levels of TfR1, DMT1 and ferritin, increased the levels of oxidation products MDA, ROS and ACSL4, decreased the level of GSH and down-regulated the levels of GPX4 and SLC7A11. The intervention of GSPE could activate Nrf2 signaling pathway to up-regulate GPX4 and SLC7A11 and increase the level of GSH. GSPE could also decrease the levels of MDA, ROS, ACSL4 and iron metabolism TfR1, DMT1, ferritin, which can improve the ferroptosis of MIN6 cells induced by high glucose and high fat. After intervention with Nrf2 siRNA, it was found that the protective effect of GSPE was inhibited and the expression of antioxidant enzymes HO-1, GPX4 and SLC7A11 was decreased. Conclusion: GSPE may reduce ferroptosis induced by high glucose and high fat by activating Nrf2 signaling pathway and up-regulating antioxidant enzymes. Consequently, the cell viability of MIN6 cells was improved.

Key words: ferroptosis, grape seed procyanidin extract (GSPE), nuclear Factor Erythroid 2-Related Factor 2 (Nrf2), MIN6

中图分类号:

R151

张丽媛刘丹丹李海燕王同玲陆恒杨瑞瑞王浩丁玉松. 葡萄籽原花青素通过Nrf2信号通路拮抗高糖高脂诱导的MIN6细胞铁死亡[J]. 食品科学, 0, (): 0-0.

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