Abstract:Objective To investigate whether the metabolite of benzene,1,4-benzoquinone(1,4-BQ),can activate PINK1/Parkin-mediated mitocphagy and the role of reactive oxygen species(ROS)in 1,4-BQ induced mitophagy in vitro.Methods Human promyelocytic leukemia cells HL60 were used as the test cells,and were divided into control group,1,4-BQ group(10 μM 1,4-BQ treated cells for 24 h),NAC group(5 mM antioxidant N-acetyl cysteine treated cells for 24 h)and 1,4-BQ+NAC group(5 mM NAC preincubated for 1 h prior to the treatment with 10 μM 1,4-BQ for 24 h). The ultra structure of the cells were observed by transmission electron microscopy(TEM),and the expression of mitophagy related protein LC3,PINK1 and Parkin were detected by Western blot,and the intracellular ROS content was determined by DCFH-DA staining.Results The mitochondria in the control group showed a normal rod-shaped structure with clear mitochondrial cristae,while in the 1,4-BQ group,the mitochondria showed a swollen structure with less mitochondrial cristae,and typical double-membrane mitophagosomes were observed. LC3-Ⅱ/LC3-Ⅰ ratio,the expression of PINK1,Parkin protein and ROS content in 1,4-BQ group were increased compared with the control group(P<0.05),and this increase was markedly blocked by the co-treatment of 1,4-BQ and NAC(P<0.05).Conclusion The 1,4-BQ can induce PINK1/Parkin-mediated mitophagy and ROS plays a significant role in 1,4-BQ-induced mitophagy.
[1] 陈玲夫,杨幼萍,李红霞,等. 温岭市制鞋业苯系物检测及暴露人群血细胞检查结果[J] . 浙江预防医学,2015,27(8):832-834. [2] SUN R,CAO M,ZHANG J,et al. Benzene Exposure Alters Expression of Enzymes Involved in Fatty Acid β-Oxidation in Male C3H/He Mice[J] . International Journal of Environmental Research and Public Health,2016,13(12):1068. [3] CHEN Y,SUN P,GUO X,et al. MiR-34a,a promising novel biomarker for benzene toxicity,is involved in cell apoptosis triggered by 1,4-benzoquinone through targeting Bcl-2[J] . Environ Pollut,2017,221:256-265. [4] FENGA C,GANGEMI S,COSTA C. Benzene exposure is associated with epigenetic changes(Review)[J] . Mol Med Rep,2016,13(4):3401-3405. [5] 吴新颖,宋淑敏,孙宝春. 新装修住宅室内空气苯系物污染状况调查[J] . 浙江预防医学,2014,26(10):1034-1035. [6] MCHALE C M,ZHANG L,SMITH M T. Current understanding of the mechanism of benzene-induced leukemia in humans:implications for risk assessment[J] . Carcinogenesis,2012,33(2):240-252. [7] BRUN S,SCHALL N,JELTSCH-DAVID H,et al. Assessing autophagy in sciatic nerves of a rat model that develops inflammatory autoimmune peripheral neuropathies[J] . Cells,2017,6(3):30. [8] EIYAMA A,OKAMOTO K. PINK1/Parkin-mediated mitophagy in mammalian cells[J] . Curr Opin Cell Biol,2015,33:95-101. [9] 覃琼玉,张亚亚,孙书强,等. 1,4-苯醌诱导 HL60 细胞自噬及活性氧在自噬发生中的作用[J] . 中华劳动卫生职业病杂志,2016,34(5):325-328. [10] 张迎梅,邱倩,漆永梅. 线粒体自噬的研究方法[J] . 兰州大学学报(自然科学版),2013,49(5):693-699. [11] 唐艳芬,阳学风. 细胞自噬与肝脏疾病关系的研究进展[J] . 中国癌症防治杂志,2017,9(2):164-166. [12] GATICA D,CHIONG M,LAVANDERO S,et al. Molecular mechanisms of autophagy in the cardiovascular system[J] . Circ Res,2015,116(3):456-467. [13] CHEN Y,SUN P,BAI W,et al. MiR-133a regarded as a potential biomarker for benzene toxicity through targeting Caspase-9 to inhibit apoptosis induced by benzene metabolite(1,4-Benzoquinone)[J] . Sci Total Environ,2016,571:883-891. [14] IVANKOVIC D,CHAU K Y,SCHAPIRA A H,et al. Mitochondrial and lysosomal biogenesis are activated following PINK1/parkin-mediated mitophagy[J] . J Neurochem,2016,136(2):388-402. [15] WEI L,WANG J,CHEN A,et al. Involvement of PINK1/ parkin-mediated mitophagy in ZnO nanoparticle-induced toxicity in BV-2 cells[J] . International Journal of Nanomedicine,2017, 12:1891-1903. [16] RAJ A,NACHIAPPAN V. Exposure to benzene metabolites causes oxidative damage in Saccharomyces cerevisiae[J] . Antonie Van Leeuwenhoek,2016,109(6):841-854. [17] UZMA N,KUMAR B S,HAZARI M A. Exposure to benzene induces oxidative stress,alters the immune response and expression of p53 in gasoline filling workers[J] . Am J Ind Med,2010,53(12):1264-1270. [18] BADHAM H J,RENAUD S J,WAN J,et al. Benzene-initiated oxidative stress:Effects on embryonic signaling pathways[J] . Chem Biol Interact,2010,184(1-2):218-221.