摘要
目的探讨纳米银对人冠状动脉内皮细胞(HCAEC)凋亡的影响及其可能的机制。方法取生长状态良好并处于对数生长期的HCAEC,分别加入浓度为0、2.5、5、10、20、40、80μg/m L的纳米银溶液,作用24 h。采用MTT法检测HCAEC凋亡率,ELISA法检测培养液上清血红素氧合酶1(HO-1)、血小板内皮细胞黏附分子1(PECAM-1)表达,火焰原子吸收光谱法检测细胞内、外银含量。结果加入2.5、5、10、20、40、80μg/m L纳米银溶液的HCAEC凋亡率分别为8.51%、21.03%、42.38%、64.62%、78.35%、74.83%。随着加入纳米银溶液浓度升高,HCAEC培养液上清HO-1表达逐渐升高,各浓度间比较P均<0.05;加入40、80μg/m L纳米银溶液的HCAEC培养液上清HO-1表达比较P>0.05。HCAEC加入10、20、40、80μg/m L纳米银溶液后细胞培养液上清PECAM-1表达均高于0、2.5、5μg/m L纳米银溶液,且加入40、80μg/m L纳米银溶液后PECAM-1表达升高更明显(P均<0.05)。加入2.5、5、10、20μg/m L纳米银溶液的HCAEC细胞内银含量低于细胞外(P均<0.01),加入40、80μg/m L纳米银溶液的HCAEC细胞内、外银含量比较差异均无统计学意义(P均>0.05)。结论纳米银可促进HCAEC凋亡,其细胞毒性作用呈浓度依赖性;通过上调HO-1、PECAM-1表达造成细胞损伤可能是其作用机制。
引文
[1]Chaloupka K,Malam Y,Seifalian AM.Nanosilver as a new generation of nanoproduct in biomedical applications[J].Trends Biotechnol,2010,28(11):580-588.
[2]Lem KW,Choudhury A,Lakhani AA,et al.Use of nanosilver in consumer products[J].Recent Pat Nanotechnol,2012,6(1):60-72.
[3]Sintubin L,Verstraete W,Boon N.Biologically produced nanosilver:current state and future perspectives[J].Biotechnol Bioeng,2012,109(10):2422-2436.
[4]Sumit A,Nikhil T,Arun B,et al.Silver nanoparticles protect human keratinocytes against UVB radiation-induced DNA damage and apoptosis:potential for prevention of skin carcinogenesis[J].Nanomedicine,2015,11(5):1265-1275.
[5]Kim SH,Ko JW,Koh SK,et al.Silver nanoparticles induce apoptotic cell death in cultured cerebral cortical neurons[J].Mol Cell Toxicol,2014,10(2):173-179.
[6]Zhang T,Wang L,Chen Q,et al.Cytotoxic potential of silver nanoparticles[J].Yonsei Med J,2014,55(2):283-291.
[7]Simkhovich BZ,Kleinman MT,Kloner RA.Air pollution and cardiovascular injury epidemiology,toxicology,and mechanisms[J].J Am Coll Cardiol,2008,52(9):719-726.
[8]Chairuangkitti P,Lawanprasert S,Roytrakul S,et al.Silver nanoparticles induce toxicity in A549 cells via ROS-dependent and ROS-independent pathways[J].Toxicol In Vitro,2013,27(1):330-338.
[9]Soto KF,Carrasco A,Powell TG,et al.Comparative in vitro cytotoxicity assessment of some manufactured nanoparticulate materials characterized by transmissionelectron microscopy[J].J Nanoparticle Res,2005,7(2):145-169.
[10]Braydich SL,Hussain SSchlager JJ,Hofmann MC.In vitro cytotoxicity of nanoparticles in mammalian germline stem cells[J].Toxicol Sci,2005,88(2):412-419.
[11]Hussain SM,Hess KL,Gearhart JM,et al.In vitro toxicity of nanoparticles in BRL 3A rat liver cells[J].Toxicol In Vitro,2005,19(7):975-983.
[12]于永生,施畅,徐丽明,等.纳米银对中国仓鼠肺成纤维细胞的细胞毒性和遗传毒性[J].癌变畸变突变,2014,26(6):428-433.
[13]Sambale F,Wagner S,Stahl F,et al.Investigations of the toxic effect of silver nanoparticles on mammalian cell lines[J].J Nanomaterials,2015,2015(1):1-9.
[14]巩凡,张帮勇,黄艳梅,等.纳米银在Hep G2和L02细胞中的生物分布及凋亡作用研究[J].环境与健康杂志,2014,31(10):893-896.
[15]黄艳梅,巩凡,张婷,等.纳米银对人肝癌细胞及正常肝细胞增殖与凋亡的影响[J].环境与健康杂志,2013,30(9):770-772.
[16]Qingwei JI,Bangwei WU,Zeng Q,et al.HO-1 attenuates atherosclerosis via regulating the imbalance of Th17/Treg in Apo E-/-mice[J].J Clin Cardiol,2016,2(1):1-4.
[17]Luo Y,Sun G,Dong X,et al.Isorhamnetin Attenuates atherosclerosis by inhibiting macrophage apoptosis via PI3K/AKT activation and HO-1 induction[J].Plos One,2015,10(3):120259.
[18]Pechlaner R,Willeit P,Summerer M,et al.Heme oxygenase-1gene promoter microsatellite polymorphism is associated with progressive atherosclerosis and incident cardiovascular disease[J].Arterioscler Thromb Vasc Biol,2015,35(1):229-236.
[19]Chistiakov DA,Orekhov AN,Bobryshev YV.Endothelial PECAM-1 and its function in vascular physiology and atherogenic pathology[J].Exp Mol Pathol,2016,100(3):409-415.
[20]Popovic'D,Starcˇevic'JN,Letonja M,et al.PECAM-1gene polymorphism(rs668)and subclinical markers of carotid atherosclerosis in patients with type 2 diabetes mellitus[J].Balkan J Med Genet,2016,19(1):63-70.
[21]Sei-ichiro T,Yuan Z and Bo Q.Heine oxygenase-l mediated cytoprotection against fiver ischemia and reperfusion injury:inhibition of type-I interferon signaling[J].Transplantation,2007,83(3):1628-1634.
