柴油废气颗粒物诱导小鼠脑微血管内皮细胞凋亡的实验研究
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摘要
目的探讨柴油废气颗粒物(diesel exhaust particles,DEP)对小鼠脑微血管内皮细胞凋亡的影响,为探讨DEP对血脑屏障的损伤作用机制提供线索。方法小鼠脑微血管内皮细胞(bEnd.3)分别用不同浓度的DEP(0、12.5、25、50和100μg/ml)处理24和48 h;用MTT比色法检测细胞活力,流式细胞术检测细胞凋亡,实时荧光定量PCR(qPCR)检测Bcl-2、Bax和Caspase-3基因表达水平,Western blot法检测Bcl-2、Bax和Caspase-3蛋白表达水平。结果分别用不同浓度DEP处理bEnd.3细胞培养24和48 h后,其中0、12.5、25、50和100μg/ml剂量组的细胞活力、细胞凋亡数量各剂量组与对照组相比,细胞活力呈下降趋势,且细胞早期凋亡数量呈上升趋势,差异有统计学意义(P<0.05)。与对照组相比,12.5、25、50和100μg/ml DEP剂量组处理24和48 h后,Bcl-2 mRNA呈下降趋势,Bax、Caspase-3 mRNA呈上升趋势,差异有统计学意义(P<0.05)。与对照组相比,24和48 h 12.5、25、50和100μg/ml DEP剂量组处理24和48 h后,Bcl-2蛋白呈下降趋势,Bax、Caspase-3蛋白呈上升趋势,差异有统计学意义(P<0.05)。结论 DEP可能通过诱导小鼠脑微血管内皮凋亡,从而对血脑屏障产生损伤作用。
引文
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[15] 张巍,金瑛,朱文赫.肽基脯氨酰顺反异构酶抑制剂胡桃醌对宫颈鳞癌SiHa细胞凋亡的作用[J].中国病理生理杂志,2015,31(3):543-546.
[16] 石元英,邓立东,饶伟文.牡荆素对人肝细胞癌SMMC-7721细胞的增殖抑制作用及其机制的影响[J].中国医院药学杂志,2016,36(5):366-371.
[17] Langford MP,Mcgee DJ,Ta KH.Multiple caspases mediate acute renal cell apoptosis induced by bacterial cell wall components[J].Ren Fail,2011,33(2):192-206.
[2] Draper W.Quantitation of nitro and dinitropolycyclic aromatic hydrocarbons in diesel exhaust particulate matter[J].Chemos,1986,15(4):437-447.
[3] Schuetzle D.Sampling of vehicle emissions for chemical analysis and biological testing.Environ[J].Environ Health Perspect,1983,47(13):65-80.
[4] Laumbach RJ,Kipen HM.Does diesel exhaust cause lung cancer (yet)[J].Am J Respir Crit Care Med,2011,183(7):843-844.
[5] Bai Y,Suzuki AK.The cytotoxic effects of diesel exhaust particles on human pulmonary artery endothelial cells in vitro:role of active oxygen species[J].Free Radical Biol Med,2001,30(5):555-562.
[6] Debra T,Sehlelf PL,Lulin JH.The Diesel exhaust in min-ers study:A nested case-control study of lung cancer and diesel exhaust[J].J Natl Cancer Inst,2012,104(11):855-868.
[7] Hartz AM,Bauer B,Block ML,et al.Diesel exhaust particles induce oxidative stress,proinflammatory signaling,and P-glycoprotein up-regulation at the blood-brain barrier[J].FASEB J,2008,22(8):2723-2733.
[8] Zha YF,Fu XZ,Zhang S.Establishment of aninvivo blood-brainbarrier model by co-culturing rat brain microvascular endothelial cell,pericytes andastrocytes[J].Chin Pharmacol Bull,2015,31(5):730-735.
[9] Deeken JF,Loscher W.The bloodbrain barrier and cancer:transporters,treatment,and Trojan horses[J].Clin Cancer Res,2007,13(6):1663-1674.
[10] 宋晓彬,范春娥,王烨.小鼠脑微血管内皮细胞的分离鉴定与培养[J].南京师大学报,2017,3(6):138-143.
[11] 张畅斌,陆茵,李沧海.小鼠脑微血管内皮细胞的培养及其生物学行为初探[J].南京中医药大学学报,2012,28(2):139-144.
[12] Wang JS,Tseng CY,Chao MW.Diesel exhaust particles contribute to endothelia apoptosis via autophagy pathway[J].Tox Sci Advance Access,2017,156(1):72-83.
[13] Klein SG,Cambier S,Hennen J,et al.Endothelial responses of the alveolar barrier in vitro in a dose-controlled exposure to diesel exhaust particulate matter[J].Part Fibre Toxicol,2017,14(1):7.
[14] 王雪芬,王磊,陈树杰.苦参素对大鼠心肌缺血再灌注损伤的保护作用及其机制研究[J].中国中医急症,2016,25(6):988-992.
[15] 张巍,金瑛,朱文赫.肽基脯氨酰顺反异构酶抑制剂胡桃醌对宫颈鳞癌SiHa细胞凋亡的作用[J].中国病理生理杂志,2015,31(3):543-546.
[16] 石元英,邓立东,饶伟文.牡荆素对人肝细胞癌SMMC-7721细胞的增殖抑制作用及其机制的影响[J].中国医院药学杂志,2016,36(5):366-371.
[17] Langford MP,Mcgee DJ,Ta KH.Multiple caspases mediate acute renal cell apoptosis induced by bacterial cell wall components[J].Ren Fail,2011,33(2):192-206.
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