TREM2基因的敲低加重纳米氧化铝导致的神经发育障碍
Knockdown of TREM2 Gene Aggravates Neural-developmental Disorders Induced by Aluminum Oxide Nanoparticles
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摘要: 纳米氧化铝（alumina nanoparticles，AlNPs）作为一种新的纳米科技材料被广泛应用，其潜在的毒性也被广泛研究。Ⅱ型髓系细胞触发受体（triggering receptor expressed on myeloid cells 2，TREM2）高度特异表达于中枢神经系统内的小胶质细胞，对脑稳态的维持及神经系统疾病发展均发挥着重要作用。为了探究trem2基因在AlNPs致神经发育障碍中的作用，将6 hpf （hours post-fertilization）的斑马鱼胚胎分为对照组（control）、阴性对照组（negative con）、TREM2敲低组（TREM2-）、纳米氧化铝组（AlNPs）、TREM2敲低+纳米氧化铝组（TREM2-/+AlNPs）。在受精1 h内用显微注射技术敲低trem2基因，并染毒144 h。染毒结束后对各组进行形态学和生殖毒性观察；用运动轨迹跟踪仪记录斑马鱼的行为轨迹。检测trem2基因、神经发育基因α1-tubulin、mbp和syn2a的表达量；以及神经递质和氧化应激情况。结果发现，各组幼鱼在死亡率、孵化率和畸形率方面无显著差异（P>0.05）。运动行为检测发现，TREM2-组、AlNPs组、TREM2-/+AlNPs组幼鱼在黑暗状态下的平均速度、移动距离、外圈停留时间百分比显著降低而绝对转角增大（P<0.05），TREM2-/+AlNPs组有明显的惊恐逃避反射（P<0.01）。此外，TREM2-组、AlNPs组和TREM2-/+AlNPs组的神经发育相关基因α1-tubulin、mbp和syn2a，乙酰胆碱酯酶（acetylcholinesterase，AChE）活性以及超氧化物歧化酶（superoxide dismutase，SOD）活性显著降低（P<0.05）。上述结果表明，AlNPs可以导致幼鱼神经发育障碍，敲低trem2基因会加重AlNPs导致的神经发育毒性，进一步说明trem2基因在神经发育中的重要作用。Abstract: As a new nanotechnology material, alumina nanoparticles (AlNPs) are widely used in our life. However, its potential toxicity has not been studied extensively. Triggering receptor expressed on myeloid cells 2 (TREM2) is highly specific expressed on microglia in the central nervous system and plays an essential role in brain homeostasis and the development of neurological diseases. To explore the role of trem2 gene in the neural-developmental toxicity induced by aluminum oxide nanoparticles, zebrafish embryos were divided into control group (control), negative control group (negative con), TREM2 knockdown group (TREM2-), AlNPs group (AlNPs), TREM2 knockdown+AlNPs group (TREM2-/+AlNPs) at 6 hours post-fertilization (hpf). The microinjection of TREM2 RNAi was completed in zebrafish eggs within 1 hour after fertilization, and they were exposed to AlNPs suspension to 144 hpf. Then, the morphology and reproduction of larvae were observed, and the behavior was tracked by an animal motion tracking instrument. The transcriptional expressions of trem2 gene and neural-developmental genes (α1-tubulin, mbp and syn2a) were detected. In addition, neurotransmitter and oxidative stress levels were measured. Our results showed that there was no significant difference in mortality, hatching rate, and deformity rate among groups (P>0.05). The average speed, moving distance, and time spent in the outer zone decreased while absolute turn angle increased in the zebrafish of the TREM2- group, AlNPs group and TREM2-/+AlNPs group (P<0.05), and panic response in the TREM2-/+AlNPs group significantly declined (P<0.01). Furthermore, the expression of α1-tubulin, mbp, and syn2a genes, and the levels of acetylcholinesterase (AChE) and superoxide dismutase (SOD) decreased (P<0.05) in TREM2- group, AlNPs group, and TREM2-/+AlNPs group. Our results suggest that AlNPs could generate neural-developmental toxicity in zebrafish, and the knockdown of trem2 gene could aggravate neurodevelopment toxicity. In summary, trem2 gene plays a vital role in neural-development.
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Key words:
- AlNPs /
- zebrafish /
- neurotoxicity /
- TREM2 /
- neural-development
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马小艺, 陈海斌. 纳米材料在生物医学领域的应用与前景展望[J]. 中国医药导报, 2006, 3(32):13-15 咸才军, 郭保文, 关延涛. 纳米材料及其技术在涂料产业中的应用[J]. 新型建筑材料, 2001, 28(5):3-5 杨龙平, 章建浩, 黄明明, 等. 纳米材料在食品包装中的应用及安全性评价[J]. 包装工程, 2015, 36(1):19-23 , 56 Yang L P, Zhang J H, Huang M M, et al. Application and safety evaluation of nanomaterial in food packaging[J]. Packaging Engineering, 2015, 36(1):19-23, 56(in Chinese)
Shrivastava R, Raza S, Yadav A, et al. Effects of sub-acute exposure to TiO2, ZnO and Al2O3 nanoparticles on oxidative stress and histological changes in mouse liver and brain[J]. Drug and Chemical Toxicology, 2014, 37(3):336-347 丁勇. 纳米氧化铝致小鼠神经发育毒性及其机制初探[D]. 太原:山西医科大学, 2015:29-32 Ding Y. The research of the developmental neurotoxicity and its mechanism induced by nano-alumina[D]. Taiyuan:Shanxi Medical University, 2015:29 -32(in Chinese)
刘焕亮, 林本成, 房彦军, 等. 纳米氧化铝致中枢神经系统和多巴胺能神经元的毒性效应研究[C]. 银川:中国毒理学会第四届中青年学者科技论坛, 2014:18-19 Gu J, Guo M, Huang C X, et al. Titanium dioxide nanoparticle affects motor behavior, neurodevelopment and axonal growth in zebrafish (Danio rerio) larvae[J]. The Science of the Total Environment, 2021, 754:142315 Wang F, Jiao C P, Liu J W, et al. Oxidative mechanisms contribute to nanosize silican dioxide-induced developmental neurotoxicity in PC12 cells[J]. Toxicology in Vitro, 2011, 25(8):1548-1556 Kleinberger G, Yamanishi Y, Su árez-Calvet M, et al.TREM2mutations implicated in neurodegeneration impair cell surface transport and phagocytosis[J]. Science Translational Medicine, 2014, 6(243):243ra86 朱婷, 孙桂波, 孟祥宝, 等. 小胶质细胞/巨噬细胞的极化及在脑卒中修复中的作用[J]. 中国药理学通报, 2019, 35(8):1046-1050 Zhu T, Sun G B, Meng X B, et al. Polarization of microglia/macrophages and its role in ischemic stroke repair[J]. Chinese Pharmacological Bulletin, 2019, 35(8):1046-1050(in Chinese)
Jadhav V S, Lin P B C, Pennington T, et al. Trem2 Y38C mutation and loss of Trem2 impairs neuronal synapses in adult mice[J]. Molecular Neurodegeneration, 2020, 15(1):62 Coe T S, Hamilton P B, Griffiths A M, et al. Genetic variation in strains of zebrafish (Danio rerio) and the implications for ecotoxicology studies[J]. Ecotoxicology, 2009, 18(1):144-150 Cao Z G, Su M L, Wang H L, et al. Carboxyl graphene oxide nanoparticles induce neurodevelopmental defects and locomotor disorders in zebrafish larvae[J]. Chemosphere, 2021, 270:128611 Dai Y J, Jia Y F, Chen N, et al. Zebrafish as a model system to study toxicology[J]. Environmental Toxicology and Chemistry, 2014, 33(1):11-17 Driever W, Stemple D, Schier A, et al. Zebrafish:Genetic tools for studying vertebrate development[J]. Trends in Genetics, 1994, 10(5):152-159 Liu K L, Petree C, Requena T, et al. Expanding the CRISPR toolbox in zebrafish for studying development and disease[J]. Frontiers in Cell and Developmental Biology, 2019, 7:13 Barros T P, Alderton W K, Reynolds H M, et al. Zebrafish:An emerging technology for in vivo pharmacological assessment to identify potential safety liabilities in early drug discovery[J]. British Journal of Pharmacology, 2008, 154(7):1400-1413 van Dycke J, Cuvry A, Knickmann J, et al. Infection of zebrafish larvae with human norovirus and evaluation of the in vivo efficacy of small-molecule inhibitors[J]. Nature Protocols, 2021, 16(4):1830-1849 Zhu J S, Xia R, Liu Z W, et al. Fenvalerate triggers Parkinson-like symptom during zebrafish development through initiation of autophagy and p38 MAPK/mTOR signaling pathway[J]. Chemosphere, 2020, 243:125336 Wu C H, Lu C W, Hsu T H, et al. Neurotoxicity of fipronil affects sensory and motor systems in zebrafish[J]. Pesticide Biochemistry and Physiology, 2021, 177:104896 Kim J, Kim C Y, Song J H, et al. Trimethyltin chloride inhibits neuronal cell differentiation in zebrafish embryo neurodevelopment[J]. Neurotoxicology and Teratology, 2016, 54:29-35 陈晓菲. 纳米ZnO对斑马鱼肝损伤及相关基因影响的研究[D]. 宁波:宁波大学, 2013:19-25 Chen X F. Damage effect of nano-ZnO on gene change of liver tissue of zebrafish (Danio rerio)[D]. Ningbo:Ningbo University, 2013:19 -25(in Chinese)
陈昱, 舒凡, 许迎春, 等. 纳米氧化钕对斑马鱼胚胎发育的毒性效应[J]. 环境与健康杂志, 2020, 37(2):122-125 , 188 Chen Y, Shu F, Xu Y C, et al. Toxic effects of nano-neodymium oxide on embryo development of zebrafish[J]. Journal of Environment and Health, 2020, 37(2):122-125, 188(in Chinese)
李晓波. 金属氧化物纳米颗粒的神经毒性研究[D]. 武汉:华中科技大学, 2008:38-51 Li X B. Neurotoxicity of metal oxide nanoparticles[D]. Wuhan:Huazhong University of Science and Technology, 2008:38 -51(in Chinese)
张勤丽, 李美琴, 徐丽, 等. 纳米氧化铝对动物神经行为改变的影响及神经细胞生存状态研究[C]. 大连:全国生化/工业与卫生毒理学学术会议论文集, 2010:268 葛翠翠, 李伟庆, 张勤丽, 等. 不同粒径纳米氧化铝对体外培养神经细胞凋亡的影响[J]. 环境与职业医学, 2012, 29(2):72-76 Ge C C, Li W Q, Zhang Q L, et al. In vitro study on the effects of nano-aluminum oxide particles with various sizes on apoptosis of neuron cells[J]. Journal of Environmental & Occupational Medicine, 2012, 29(2):72-76(in Chinese)
陈金, 范蓉, 张萍, 等. 纳米氧化铝对斑马鱼幼鱼的神经毒性及mTOR基因的作用[J]. 环境与职业医学, 2019, 36(5):431-437 Chen J, Fan R, Zhang P, et al. Al2O3 nanoparticles induced neurotoxicity and role of mTOR gene in zebrafish larvae[J]. Journal of Environmental and Occupational Medicine, 2019, 36(5):431-437(in Chinese)
王艳红. mTOR基因在纳米氧化铝诱导的斑马鱼生长发育障碍中的作用研究[D]. 太原:山西医科大学, 2021:16-31 Wang Y H. Role of mTOR gene in the growth and development of zebrafish induced by aluminum oxide nanoparticles[D]. Taiyuan:Shanxi Medical University, 2021:16 -31(in Chinese)
陈金. 纳米氧化铝胚胎暴露致成年斑马鱼的学习和记忆能力进行性损伤[D]. 太原:山西医科大学, 2020:15-32 Chen J. Progressive impairment of learning and memory in adult zebrafish treated by Al2O3
nanoparticles when in embryos[D]. Taiyuan:Shanxi Medical University, 2020:15-32(in Chinese)黄涛. PIK3C3/Vps34基因调控的自噬在纳米氧化铝致斑马鱼幼鱼神经发育毒性中的作用[D]. 太原:山西医科大学, 2021:13-26 Huang T. Role of autophagy regulated by PIK3C3 /Vps34 gene in neurodevelopmental toxicity of zebrafish larvae induced by AlNPs[D]. Taiyuan:Shanxi Medical University, 2021:13-26(in Chinese)
Wang Y M, Cella M, Mallinson K, et al.TREM2lipid sensing sustains the microglial response in an Alzheimer's disease model[J]. Cell, 2015, 160(6):1061-1071 李凌杰, 于晓琳, 刘瑞田. 胶质细胞介导的神经突触修剪在阿尔茨海默病中的作用[J]. 药学学报, 2021, 56(2):383-390 Li L J, Yu X L, Liu R T. Synaptic pruning mediated by glia in Alzheimer's disease[J]. Acta Pharmaceutica Sinica, 2021, 56(2):383-390(in Chinese)
Neumann H, Takahashi K. Essential role of the microglial triggering receptor expressed on myeloid cells-2(TREM2) for central nervous tissue immune homeostasis[J]. Journal of Neuroimmunology, 2007, 184(1-2):92-99 Paloneva J, Autti T, Raininko R, et al. CNS manifestations of Nasu-Hakola disease:A frontal dementia with bone cysts[J]. Neurology, 2001, 56(11):1552-1558 Paloneva J, Manninen T, Christman G, et al. Mutations in two genes encoding different subunits of a receptor signaling complex result in an identical disease phenotype[J]. The American Journal of Human Genetics, 2002, 71(3):656-662 Paloneva J, Kestilä M, Wu J, et al. Loss-of-function mutations in TYROBP (DAP12) result in a presenile dementia with bone cysts[J]. Nature Genetics, 2000, 25(3):357-361 Guo Y, Wei X B, Yan H, et al.TREM2deficiency aggravates α-synuclein-induced neurodegeneration and neuroinflammation in Parkinson's disease models[J]. FASEB Journal:Official Publication of the Federation of American Societies for Experimental Biology, 2019, 33(11):12164-12174 胡占英, 张靖溥. 长春新碱对斑马鱼神经发育和行为的影响[J]. 毒理学杂志, 2014, 28(2):98-103 Hu Z Y, Zhang J P. Effects of vincristine on zebrafish neurodevelopment and behavior[J]. Journal of Toxicology, 2014, 28(2):98-103(in Chinese)
Song Y Y, Kirkpatrick L L, Schilling A B, et al. Transglutaminase and polyamination of tubulin:Posttranslational modification for stabilizing axonal microtubules[J]. Neuron, 2013, 78(1):109-123 Jung S H, Kim S, Chung A Y, et al. Visualization of myelination in GFP-transgenic zebrafish[J]. Developmental Dynamics:An Official Publication of the American Association of Anatomists, 2010, 239(2):592-597 Fan C Y, Cowden J, Simmons S O, et al. Gene expression changes in developing zebrafish as potential markers for rapid developmental neurotoxicity screening[J]. Neurotoxicology and Teratology, 2010, 32(1):91-98 何梦婷, 王伟, 许洁, 等. 氧化应激参与溴乙酰胺致斑马鱼胚胎神经发育毒性[J]. 环境与职业医学, 2021, 38(6):586-592 He M T, Wang W, Xu J, et al. Role of oxidative stress in neurodevelopmental toxicity of bromoacetamide in zebrafish embryos[J]. Journal of Environmental and Occupational Medicine, 2021, 38(6):586-592(in Chinese)
Fu J J, Guo Y Y, Yang L H, et al. Nano-TiO2 enhanced bioaccumulation and developmental neurotoxicity of bisphenol A in zebrafish larvae[J]. Environmental Research, 2020, 187:109682 张立敏, 顾超, 安红梅. 氧化应激介导的细胞凋亡在阿尔茨海默病中的作用[J]. 医学综述, 2021, 27(9):1685-1690 Zhang L M, Gu C, An H M. Role of oxidative stress-mediated apoptosis in Alzheimer's disease[J]. Medical Recapitulate, 2021, 27(9):1685-1690(in Chinese)
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