波动性葡萄糖对胰岛β细胞INS-1的影响  

Effect of fluetuant high glucose to pancreatic β-Cell lines INS-1

作　　者：龙可[1] 薛耀明[1] 沙建平[1] 桑丹[1] 林占[1] 

机构地区：[1]南方医科大学南方医院内分泌代谢科,广州510515

出　　处：《中华胰腺病杂志》2009年第2期118-120,共3页Chinese Journal of Pancreatology

摘　　要：目的探讨波动性葡萄糖对培养的胰岛β细胞株INS-1细胞的损伤机制。方法实验分5组。对照组：葡萄糖浓度为5．5mmol／L；持续高糖组：葡萄糖浓度为16．7mmol／L；波动性高糖组：16．7mmo／L葡萄糖培养2h后更换葡萄糖浓度为5．5mmo／L，继续培养3h，每天重复3次，夜间9h维持在5．5mmo／L葡萄糖的培养基中；N-乙酰半胱氨酸（NAC，1．0mmo／L）＋高糖组；NAC＋波动性高糖组。流式细胞仪检测活性氧簇（ROS）的水平；四氮唑蓝定量检测细胞内葡萄糖-6-磷酸脱氢酶（G6PD）的活性；同时检测还原型辅酶Ⅱ（NADPH）的含量。结果干预72h后，对照组、高糖组、波动性高糖组、NAC＋高糖组和NAC＋波动性高糖组细胞的ROS活性分别为37．77±2．31、86．97±7．97、124．27±10．04、60．92±2．61和51．47±3．36；G6PD活性分别为1．25±0．03、1．09±0．02、1．03±0．01、1．12±0．02和1．21±0．01；NADPH含量分别为（0．123±0．003）mmol／mgprot、（0．112±0．004）mmol／mgprot、（0．099±0．002）mmol／mgprot、（0．116±0．005）mmol／mgprot和（0．120±0．002）mmol／mgprot。波动性高糖组细胞ROS活性较单纯高糖组显著增加（P〈0．01），G6PD活性和NADPH含量较单纯高糖组显著减少（P〈0．01）；加NAC共孵育使细胞的变化程度减小。结论波动性高浓度葡萄糖使培养的INS-1细胞氧化应激水平增高，其机制可能是由于抗氧化酶G6PD活性降低导致了氧化还原的失衡。Objective To investigate the damage mechanism of fluctuant high glucose on the INS-1 cells （pancreatic β-cell lines）. Methods The cells were divided into five groups： the control groups （A group： 5.5 mmol/L of glucose）, the continuing high glucose group （ B group： 16.7 mmol/L of glucose） , the fluctuant glucose group （ C group： 16.7 mmol/L of glucose for cultivation for 2 h, then the concentration changed to 5.5 mmol/L for cultivation for 3 h, which was repeated 3 times per day; the cells were kept in the medium containing 5.5 mmol/L of glucose during night time for 9 h） , the continuing high glucose plus NAC （1.0 mmo/L） group （D group）, the fluctuant glucose plus NAC group （E group）. The intracellular reactive oxygen species （ROS） was observed by the flow cytometry. The activity of glucose-6-phosphate dehydrogenase （G6PD） was estimated by the tetrazolium linked cytochemical method. Results 72 h after intervention, the levels of ROSwere 37.77 ± 2.31, 86.97 ±7.97, 124.27±10.04, 60.92 ±2.61 and 51.47±3.36, respectively, in A - E group; the activities of G6PD were 1.25 ±0.03, 1.09 ±0.02, 1.03 ±0.01, 1.12 ± 0.02 and 1.21 ± 0.01, respectively; the levels of NADPH were （0. 123 ±0.003） mmol/mg prot, （0. 112 ± 0.004） retool/rag prot, （0. 099 ± 0. 002 ） retool/mg prot, （ 0. 116 ±0. 005 ） retool/rag prot and （ 0. 120±0.002） mmol/mg prot, respectively. The level of ROS in the cells of the fluctuant glucose group were significantly higher than that in the continuing high glucose group （P 〈 0.01 ）. The G6PD activity and NADPH was significantly lower in fluetuant high glucose group than those in the continuing high glucose group （ P 〈0.01）. NAC co-cultivation decreased the extent of cell＇s change. Conclusions Exposure of INS-1 to high glucose lead to increased oxidative stress, possible mechanism included decreased G6PD activity and subsequent imbalance between oxidation and reduction.

关 键 词：胰腺Β细胞 活性氧 葡萄糖-6-磷酸脱氢酶 波动性高糖 

分 类 号：R587.1[医药卫生—内分泌] R446.19[医药卫生—内科学]