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

发布时间：2018-01-03 19:42

本文关键词：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　出处：《中国微生态学杂志》2015年10期 　论文类型：期刊论文

【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(P0.01)。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。
[Abstract]:Objective To observe the effect of Pseudomonas aeruginosa exotoxin A (PEA) effect on MLE-12 cell oxidative stress and autophagy activity. MLE-12 cells were cultured to the logarithmic growth phase in vitro, respectively using PEA 0200400800 and 1H 1 cells treated with 000ng/mL and 2H, then further using PEA 1 cells treated with 000ng/mL 15min, 30min, 1H, 2H and 3H the generation of hydrogen peroxide, kit for detection of H2O2, detection of microtubule associated protein 1 light chain 3 (Western-blot LC3) expression ratio analysis of LC3II and beta -actin. Results compared with the control group cells, with the increase of the concentration of PEA, cell H2O2 detection was gradually increased after 2H treatment, PEA H2O2, the cells were detected respectively is 1.73,2.09,3.82 and 5.06 times higher than the control group (P0.01). At the same time, the expression of autophagy marker LC3II in cells increased significantly. In addition, the high concentration of PEA (1 000ng/mL) after treating cells, With the prolongation of time, the detection of H2O2 increased significantly, accompanied by the enhancement of autophagy. Conclusion PEA can induce oxidative stress damage and activate autophagy in MLE-12 cells.

【作者单位】：中国医科大学附属第一医院老年医学科;中国医科大学附属第一医院血液内科;中国医科大学附属盛京医院第二肿瘤病房;
【分类号】：R363
【正文快照】： 铜绿假单胞菌(Pseudomonas aeruginosa,PA)是一种革兰阴性条件致病菌,常见于肺间质纤维化、肿瘤、免疫功能缺陷患者的肺部感染。由于其复杂的发病机制及对抗生素的广泛耐药性,PA感染的临床治疗面临着长期挑战。细胞自噬作为机体的一种防御机制,在感染、免疫过程以及炎症、肿瘤

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