CAJ | 학술논문

探讨糖化终末产物(AGE)对小鼠胰岛细胞株MIN6细胞活力及活性氧(ROS)水平的影响.制备BSA-AGE,用不同浓度AGE(100、200、400 mg/L)干预MIN6细胞不同时间后,MTF比色法检测细胞活力变化.以活性氧捕获剂双氢-乙酰乙酸二氯荧光黄(DCFH-DA)孵育细胞,通过流式细胞仪检测细胞内二氯荧光黄(DCF)的荧光强度而测得细胞内活性氧水平,并测定胰岛素分泌的变化.随着AGE浓度的升高和作用时间的延长,细胞活力明显下降(P＜0.05).经DCFH-DA孵育后流式细胞仪检测显示,AGE处理组细胞内DCF平均荧光强度较对照组明显升高(P＜0.05).胰岛素分泌量随着AGE浓度的增高和时间的延长,呈下降趋势(P＞0.05).提示BSA-AGE抑制MIN6活力,使细胞内活性氧生成增加,诱导MIN6细胞氧化应激.
탐토당화종말산물(AGE)대소서이도세포주MIN6세포활력급활성양(ROS)수평적영향.제비BSA-AGE,용불동농도AGE(100、200、400 mg/L)간예MIN6세포불동시간후,MTF비색법검측세포활력변화.이활성양포획제쌍경-을선을산이록형광황(DCFH-DA)부육세포,통과류식세포의검측세포내이록형광황(DCF)적형광강도이측득세포내활성양수평,병측정이도소분비적변화.수착AGE농도적승고화작용시간적연장,세포활력명현하강(P＜0.05).경DCFH-DA부육후류식세포의검측현시,AGE처리조세포내DCF평균형광강도교대조조명현승고(P＜0.05).이도소분비량수착AGE농도적증고화시간적연장,정하강추세(P＞0.05).제시BSA-AGE억제MIN6활력,사세포내활성양생성증가,유도MIN6세포양화응격.
To explore the effect of advanced glycation end-products(AGEs)on cell viability and level of reactive oxygen species(ROS)in MIN6 cells. After intervention of various concentrations(100,200, and 400 mg/L)of AGEs for some time, cell viability was detected by MTT assay. 2', 7'-dichlorofluorescein diacetate(DCFH-DA)was used as a reactive oxygen species capture agent. The fluorescent intensity of 2', 7'-dichlorofluorescein(DCF), which was the product of cellular oxidation of DCFH-DA, was detected by flow cytometry. The level of ROS and insulin secretion was thus measured. Viability of MIN6 cells was inhibited by AGEs in a dose and time dependent manner(P＜0.05).Intracellular fluorescent intensity of DCF was markedly elevated in the AGEs groups as compared with that in the control group(P＜0.05).Insulin secretion was decreased in the AGEs groups than that in the control group(P＞0.05). The results suggest that AGEs inhibit the viability and induce oxidative stress in MIN6 cells by overproduction of ROS.