SOD对青石棉诱导BEAS-2B细胞产生过氧化物歧化酶的作用

Crocidolite induces ROS generation and SOD blocking effect in BEAS-2B cells

目的:探讨SOD对青石棉诱导BEAS-2B细胞产生过氧化物歧化酶(ROS)的抑制作用。方法:采用台盼蓝吞噬实验和乳酸脱氢酶同工酶(LDH)试剂盒分别测定0mg/L、100mg/L、250mg/L、500mg/L青石棉作用后BE-AS-2B细胞成活率和培养上清LDH水平。分别采用双氢溴乙啶(DHE)和双氯荧光素双醋酸盐(DCF)荧光探针测定青石棉刺激BEAS-2B细胞后ROS的产生情况,同时使用200USOD与青石棉共孵育观察其对ROS产生的抑制作用。结果:100mg/L以下剂量的青石棉与BEAS-2B细胞共培养24h后细胞成活率为97%,未显示明显毒性;DHE荧光染色显示青石棉可诱导BEAS-2B细胞产生ROS,细胞呈深红染色,使用SOD后可抑制ROS产生,细胞染色减弱;且不同剂量青石棉作用BEAS-2B细胞不同时间后DCF染色差异无统计学意义(P>0.05)。结论:青石棉可诱导BEAS-2B细胞产生ROS,且该作用可被SOD抑制。

Aim: To observe the ROS generation and SOD inhibition in BEAS-2B cells induced by crocidolite and discuss the pathogenesis of crocidolite-related diseases. Methods: Trypan Blue phagocytosis was used to evaluate the morphology of the cells stimulated with 0 mg/L,100 mg/L,250 mg/L and 500 mg/L crocidolite. LDH kit was employed to detect the releasing of LDH in the cultured supernatant. ROS was detected using DHE and DCF fluorescence probes, at the same time, SOD reagent was employed to observe its blocking effect on ROS generation. Results: Trypan Blue phagocytosis test showed that more than 97% of the BEAS GAAB2 2B cells cocultured with crocidolite could uptake the staining below the dose of 100 mg/L. Cytoxicity assay showed no obviously toxic effect using crocidolite at the dose of 100 mg/L for 24 hours incubating with BEAS GAAB2 2B cells. DHE staining indicated that the deep staining in crocidolite treated cells was much stronger than that of the cells without exposure to crocidolite, however, the deep staining could be reduced by adding SOD to the cell plate. DCF staining showed that the OD values in cells exposed to different concentration of crocidolite for different time had no significant difference (P>0.05). Conclusion: ROS generation could be induced by crocidolite in BEAS-2B cells,which can be inhibited by SOD.