PFOS对PC12细胞自噬与凋亡的影响Effect of PFOS on Autophagy and Apoptosis in PC12 Cells
刘晓晖;李振伟;刘畅;朱春艳;钱波;邵静;胡宏;
摘要(Abstract):
为了研究新型持久性有机污染物全氟辛烷磺酸(peruorooctane sulfonate,PFOS)的神经毒性及其机制,将PC12细胞暴露于31.25、62.5、125、250、500μmol·L-1的PFOS,利用噻唑蓝(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT)法检测细胞存活率、二氯二氢荧光素-乙酰乙酸酯(2’,7’-Dichlorofluorescin diacetate,DCFH-DA)荧光探针检测活性氧含量、Hoechst 33342/PI试剂盒检测细胞凋亡及坏死情况、溶酶体特异性荧光染料Lyso-Tracker Red监测细胞内溶酶体数量;利用实时荧光定量PCR(quantitative real-time reverse transcription-polymerase chain reaction,q RT-PCR)技术对自噬发生的代表性基因Atg5、Atg 12、Beclin 1的表达情况进行考察。结果发现,与对照组比较,250、500μmol·L-1PFOS显著降低PC12细胞存活率(P<0.05);62.5、125、250、500μmol·L-1PFOS诱导活性氧含量升高(P<0.05);62.5、125、250、500μmol·L-1PFOS诱导PC12细胞凋亡(P<0.05);125、250、500μmol·L-1PFOS升高溶酶体数量及Atg 5、Atg 12、Beclin 1基因表达(P<0.05)。结果表明,PFOS能够诱导PC12细胞死亡,其机制可能为干扰自噬与凋亡过程。
关键词(KeyWords): PFOS;PC12;神经毒性;凋亡;自噬
基金项目(Foundation): 国家自然科学基金(81273031;81302400);; 大连理工大学工业生态与环境工程教育部重点实验室开放基金(KLIEEE-12-04)
作者(Author): 刘晓晖;李振伟;刘畅;朱春艳;钱波;邵静;胡宏;
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参考文献(References):
- [1]Saikat S,Kreis I,Davies B,et al.The impact of PFOS on health in the general population:A review[J].Environmental Science:Processes&Impacts,2013,15(2):329-335
- [2]Eriksen K T,Raaschou-Nielsen O,Mc Laughlin J K,et al.Association between plasma PFOA and PFOS levels and total cholesterol in a middle-aged Danish population[J].Plo SOne,2013,8(2):e56969
- [3]Slotkin T,Mac Kli op E,Melnick R,et al.Developmental neurotoxicity of perfluorinated chemicals modeled in vitro[J].Environmental Health Perspectives,2008,116(6):716-722
- [4]Butenhoff J L,Chang S C,Ehresman D J,et al.Evaluation of potential reproductive and developmental toxicity of potassium perfluorohexanesulfonate in Sprague Dawley rats[J].Reproductive Toxicology,2009,27(3):331-341
- [5]Chen M,Ha E,Liao H,et al.Perfluorinated compound levels in cord blood and neurodevelopment at 2 years of age[J].Epidemiology,2013,24(6):800-808
- [6]Lee H G,Lee Y J,Yang J H.Perfluorooctane sulfonate induces apoptosis of cerebellar granule cells via a ROS-dependent protein kinase C signaling pathway[J].Neurotoxicology,2012,33(3):314-320
- [7]Li F,Li X,Luo Y,et al.Autophagy inhibition induces podocyte apoptosis by activating the pro-apoptotic pathway of endoplasmic reticulum stress[J].Experimental Cel Research,2014,322(2):290-301
- [8]许秋菊,侯莉莉,胡国强,等.麦冬皂苷B诱导人宫颈癌He La细胞自噬的机制[J].药学学报,2013,48(6):855-859Xu Q,Hou L,Hu G,et al.Molecular mechanism of ophiopogonin B induced cellular autophagy of human cervicalcancer He La cells[J].Acta Pharmaceutica Sinica,2013,48(6):855-859(in Chinese)
- [9]Muoz M A,Pacheco A,Becker M I,et al.Different cell death mechanisms are induced by a hydrophobic flavin in human tumor cells after visible light irradiation[J].Journal of Photochemistry and Photobiology B:Biology,2011,103(1):57-67
- [10]Ceccatelli S,DaréE,Moors M.Methylmercury-induced neurotoxicity and apoptosis[J].Chemico-Biological Interactions,2010,188(2):301-308
- [11]Xu G F,Zhou Q,Wan C H,et al.2,3,7,8-TCDD induces neurotoxicity and neuronal apoptosis in the rat brain cortex and PC12 cell line through the down-regulation of the Wnt/β-catenin signaling pathway[J].Neuro Toxicology,2013,37:63-73
- [12]Wang X F,Zhao G P,Liang J T,et al.PFOS-induced apoptosis through mitochondrion-dependent pathway in human-hamster hybrid cells[J].Mutation Research/Genetic Toxicology and Environmental Mutagenesis,2013,754(1-2):51-57
- [13]Yu Y,Duan J,Yu Y,et al.Silica nanoparticles induce autophagy and autophagic cell death in Hep G2 cells triggered by reactive oxygen species[J].Journal of Hazardous Materials,2014,270:176-186
- [14]Deng X,Zhang F,Rui W,et al.PM2.5-induced oxidative stress triggers autophagy in human lung epithelial A549 cells[J].Toxicology in Vitro,2013,27(6):1762-1770
- [15]邓琦程,王福萍,高爱华,等.Beclinl和Atg5在上皮性卵巢癌组织中的表达及临床意义[J].基础医学与临床,2013,33(12):1595-1599Deng Q,Wang F,Gao A,et al.Expression and clinical significance of Beclin 1 and Atg 5 in epithelial ovarian cancer tissue[J].Basic and Clinical Medicine,2013,33(12):1595-1599(in Chinese)
- [16]张发仁,许丽艳,沈忠英,等.自噬的概述[J].汕头大学医学院学报,2005,18(4):250-253
- [17]Eisenberg-Lerner A,Bialik S,Simon H,et al.Life and death partners:Apoptosis,autophagy and the cross-talk between them[J].Cel Death&Differentiation,2009,16(7):966-975
- [18]Yang L J,Chen Y,He J,et al.Betulinic acid inhibits autophagicux and induces apoptosis in human multiple myeloma cells in vitro[J].Acta Pharmacologica Sinica,2012,33(12):1542-1548
- [19]Yao X F,Cao J,Xu L M,et al.Perfluorooctane sulfonate blocked autophagy flux and induced lysosome membrane permeabilization in Hep G2 cells[J].Food and Chemical Toxicology,2014,67:96-104