铜绿假单胞菌外毒素A诱导MLE-12细胞氧化应激损伤并激活自噬  

Paeruginosa exotoxin A induced oxidative stress triggers autophagy in mouse lung epithelial 12 cells

作　　者：王婷婷[1] 康思静 胡天玉[3] 

机构地区：[1]中国医科大学附属第一医院老年医学科,辽宁沈阳110001 [2]中国医科大学附属第一医院血液内科,辽宁沈阳110001 [3]中国医科大学附属盛京医院第二肿瘤病房,辽宁沈阳110001

出　　处：《中国微生态学杂志》2015年第10期1148-1151,共4页Chinese Journal of Microecology

摘　　要：目的观察铜绿假单胞菌外毒素A(PEA)对MLE-12细胞的氧化应激损伤及其自噬活动的影响。方法体外培养MLE-12细胞至对数生长期,分别采用PEA 0、200、400、800和1 000ng/mL处理细胞1h和2h,之后进一步采用PEA 1 000ng/mL处理细胞15min、30min、1h、2h和3h,应用过氧化氢试剂盒检测H2O2的生成量,Western-blot法检测微管相关蛋白1轻链3(LC3)的表达,分析LC3II与β-actin之间的比值变化。结果与对照组细胞相比较,随着PEA浓度的增高,细胞的H2O2检出量逐渐增加,PEA处理2h后,各组细胞H2O2检出量分别是对照组的1.73、2.09、3.82和5.06倍(P<0.01)。与此同时,细胞中自噬标记物LC3II的表达量显著增加。此外,高浓度PEA(1 000ng/mL)处理细胞后,随着时间的延长,H2O2检出量明显升高,同时伴随着细胞自噬活动的增强。结论 PEA可以诱导MLE-12细胞产生氧化应激损伤,并激活细胞的自噬活动。Objective To investigate the influences of Paeruginosa exotoxin A （PEA） on oxidative stress inju- ry and autophagy activity of MLE-12 cells. Methods MLE-12 cells in logarithmic phase were treated with PEA at different concentrations for 1 or 2 hours. Furthermore, the MLE-12 cells were exposed to PEA at the concentration of 1,000 ng/mL for different length of time. The production of H2 O2 was quantified with Hydrogen Peroxide Assay Kit. The expression of LC3 protein was examined by Western blot and LC3 Ⅱ/β-actin ratio was measured, respectively. Results Compared with the control group, the PEA treated ceils showed significant increases of H2 O2 with the increased concentration of PEA. The production of H2O2 were increased by 1.73 times, 2.09 times, 3.83 times and 5.06 times respectively compared with the control group after stimulated by PEA for 2 hours （P〈0.01）. At the same time, the expression of autophagy marker LC3 Ⅱ was also significantly increased. Moreover, there was a significant rise in both H202 and LC3Ⅱ after the cells were treated with high concentration of PEA （1,000 ng/mL） along with the time ex- tended. Conclusion PEA can induce oxidative stress injuries and activate cell autophagy activity in MLE-12 cells.

关 键 词：铜绿假单胞菌外毒素A 氧化应激 自噬 

分 类 号：Q782[生物学—分子生物学]