摘要
目的大气细颗粒所含的过渡金属元素与呼吸道疾病的发生密切相关,但其诱导的氧化应激对呼吸道细胞的影响仍不十分清楚。本文主要研究过渡金属元素Fe诱导的肺泡上皮细胞氧化应激对肺表面蛋白C(SP-C)表达的影响。方法肺泡II型上皮MLE-12细胞经Fe暴露处理24 h后,使用噻唑蓝(MTT)、2,7-二氯荧光素(DCF)和流式细胞测定细胞活力、细胞内活性氧(ROS)生成和细胞凋亡;采用实时定量PCR和Western blot分析细胞的SP-C mRNA和蛋白表达水平变化。结果 Fe明显抑制MLE-12细胞活力(P<0.01),升高细胞内的ROS生成水平(P<0.01),并上调细胞SP-C mRNA和蛋白的表达水平。但其表达水平被过氧化氢酶、乙酰-L-半胱氨酸和磺酸去铁敏预处理后明显降低。结论 Fe诱导生成的ROS参与SP-C的表达。
Objective To investigate the effect of transitional element iron（ Fe）-induced oxidative stress on the expression of surfactant protein C（ SP-C） in MLE-12 cells. Methods MLE-12 cells were treated with Fe Cl3 at a concentration of 100 μmol / L for 24 h.Cell viability was determined by MTT assay. Intracellular reactive oxygen species（ ROS） generation was detected using 2,7-dichlorofluorescein diacetate（ DCFH-DA）. The number of apoptotic cells was measured by flow cytometry. The expressions of SP-C mRNA and protein were detected by using real time-PCR and Western blot. Results Fe Cl3 significantly decreased cell viability and triggered an increase of intracellular reactive oxygen species（ ROS） and apoptosis（ P 〈 0. 01）. The data of real time-PCR and Western blot showed that Fe Cl3 treatment up-regulated the expressions of SP-C mRNA and protein in MLE-12 cells（ P 〈 0. 01）. However,the expressions of SP-C mRNA and protein were significantly down-regulated by pretreatment with catalase（ CAT）,N-acetylcysteine（ NAC） and deferoxamine（ DFO）. Conclusion These results suggest that Fe Cl3-induced ROS might function as signaling molecules to up-regulate SP-C expression.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2015年第1期15-18,共4页
Journal of Toxicology
基金
国家自然科学基金委面上项目(21377127
11275264
U1432245
J1103520)
