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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 The transition metal elements contained in the fine particles of the air are closely related to the occurrence of respiratory diseases, but the effects of oxidative stress on the airway cells are still not very clear. In this paper, we investigated the effect of transition metal element Fe-induced alveolar epithelial cell oxidative stress on the expression of lung surface protein C (SP-C). Methods Alveolar type II epithelial MLE-12 cells were exposed to Fe for 24 h. MTT, DCF and flow cytometry were used to determine the cell viability, intracellular reactive oxygen species (ROS) Generation and apoptosis. The changes of SP-C mRNA and protein expression were analyzed by real-time PCR and Western blot. Results Fe significantly inhibited the viability of MLE-12 cells (P <0.01), increased ROS production (P <0.01) and up-regulated the expression of SP-C mRNA and protein. But its expression level was significantly reduced by catalase, acetyl-L-cysteine and desferrioxamine sulfonate. Conclusion Fe-induced ROS is involved in the expression of SP-C.