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目的探究IR对PC12细胞缺氧/复氧损伤的影响,及抗氧化剂N-乙酰-L-半胱氨酸(NAC)对IR PC12细胞缺氧/复氧损伤的保护作用。方法采用100nmol/L胰岛素诱导PC12细胞产生IR,Na_2S_2O_4建立PC12细胞缺氧/复氧模型,CCK-8法检测细胞活力,葡萄糖氧化酶法检测培养液上清中葡萄糖含量并计算葡萄糖消耗量,黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性,硫代巴比妥酸法测定丙二醛(MDA)含量,流式细胞术分别检测细胞凋亡和线粒体膜电位。结果高胰岛素可在不影响PC12细胞活力的情况下抑制胰岛素诱导的葡萄糖摄取(P<0.05);在IR PC12细胞中,SOD活性较低,MDA水平较高(P<0.05);IR可加重缺氧/复氧诱导的PC12细胞凋亡和线粒体膜电位去极化(P<0.05);NAC可抵消IR诱导的上述作用(P<0.05)。结论 IR可通过增强氧化应激加重PC12细胞缺氧/复氧损伤,而NAC可通过抑制氧化应激和稳定线粒体膜电位,减轻IR PC12细胞缺氧/复氧损伤。
Objective To investigate the effect of IR on hypoxia / reoxygenation injury in PC12 cells and the protective effect of N-acetyl-L-cysteine (NAC) on hypoxic / reoxygenation injury in IRPC12 cells. Methods The PC12 cells were induced by 100nmol / L insulin to induce IR and Na_2S_2O_4 to establish hypoxia / reoxygenation model of PC12 cells. Cell viability was detected by CCK-8 assay. Glucose oxidase activity was measured by glucose oxidase method and the glucose consumption was calculated. The activity of superoxide dismutase (SOD) was measured by purine oxidase method. The content of malondialdehyde (MDA) was determined by thiobarbituric acid method. The apoptosis and mitochondrial membrane potential were detected by flow cytometry. Results Insulin could inhibit insulin-induced glucose uptake (P <0.05) without affecting the viability of PC12 cells. In IR PC12 cells, SOD activity was lower and MDA level was higher (P <0.05) Oxygen / reoxygenation-induced PC12 cell apoptosis and mitochondrial membrane potential depolarization (P <0.05); NAC can counteract the above-mentioned effects induced by IR (P <0.05). Conclusion IR can increase the hypoxia / reoxygenation injury in PC12 cells by increasing oxidative stress. However, NAC attenuates hypoxia / reoxygenation injury in IRPC12 cells by inhibiting oxidative stress and stabilizing mitochondrial membrane potential.