孕期亚硝酸盐暴露对子鼠视皮质突触改变的影响

doi:10.16098/j.issn.0529-1356.2015.06.002

摘要/Abstract

摘要：

目的建立亚硝酸盐暴露模型，探讨孕期亚硝酸盐暴露对子鼠视皮质突触数量的影响及其改变机制。方法采用免疫荧光染色法观察对照组和模型组（低剂量组与高剂量组）子鼠出生后0d(P0)、P7、P14、P30的的视皮质突触素(SYN)和γ突触核蛋白(γ-synuclein)的改变，利用Western blotting和水迷宫实验对SYN蛋白的表达进行进一步验证。结果横向比较（剂量）发现，亚硝酸盐暴露组SYN减少(P<0.05)而γ-synuclein 阳性细胞增多(P<0.05)，且与亚硝酸盐存在剂量依赖关系(P<0.05)。纵向比较(年龄)发现，P0、P7、P14、P30 SYN表达随着年龄增长而增多(P<0.05),γ-synuclein表达减少 (P<0.05)，剂量(MS=2.00, F=55.22,P<0.05)与年龄(MS=1.11, F=30.68, P<0.05)之间存在交互作用且剂量的影响作用更大。Western blotting和水迷宫实验结果与免疫荧光统计结果一致。结论孕期亚硝酸盐暴露会影响子代视皮质突触的发育，减少突触数量，且对其影响与亚硝酸盐暴露剂量存在依赖关系，而γ突触核蛋白的过度表达可能是突触丢失的主要原因。

关键词: 孕期亚硝酸盐暴露, 突触, γ突触核蛋白, 视皮质, 免疫印迹法, 小鼠

Abstract:

Objective To understand the nitrite’s toxicity to the quantitative alternations of synapses and explore its possible mechanism in mouse visual cortex using prenatal nitrite exposure(PNE) models. Methods The pregnant dams were exposure to nitrite from E0 (embryonic day 0) to parturition and divided into control group, moderate dose group (60mg/kg) and high dose groups (120mg/kg). Pups with different treatments at P0, P7, P14 and P30 were harvested. Synaptophysin expression and γ-synucein expression in visual cortex were visualized with immunocytochemistry technique and Western blotting. Synaptophysin positive puncta and γ-synuclein positive neurons were measured, and their correlation was tested with statistical analysis. Results The pups’learning and memory reduced after nitrite exposure. The synaptophysin positive buttons reduced after prenatal nitrate exposure with dose dependency (P<0.05), and the γ-synuclein positive cells increased with dose dependency (P<0.05) as well. There was negative correlation between synaptophysin expression and γ-synuclein expression. Western blotting supported the results from immunofluorescent labeling. Conclusion Prenatal nitrite exposure can induce the synapse loss with dose dependency and long-term effect, and the abnormal aggregation of γ-synuclein may contribute to the synaptic loss.

Key words: Prenatal nitrite exposure, Synapse, γ-synuclein, , Visual cortex, , Western blotting, Mouse

李瑞玲陈永芳王颖董红静时书勤邓锦波* 邓洁心* . 孕期亚硝酸盐暴露对子鼠视皮质突触改变的影响[J]. 解剖学报, 2015, 46(6): 729-736.

LI Rui-ling CHEN Yong-fang WANG Ying DONG Hong-jing SHI Shu-qin DENG Jin-bo*DENG Jie-xin*. Synapse loss in visual cortex of mouse after prenatal nitrite exposure[J]. AAS, 2015, 46(6): 729-736.

参考文献

［1］Saigal R, Goyal LK, Agrawal A, et al. Methaemoglobinaemia as a result of nitrite poisoning［J］. J Assoc Physicians India, 2014, 62(8): 737-738.
［2］Lu X, Lai Z, Du MJ. Protection of Salvia miltiorrhiza against acute sodium nitrite poisoning in mice［J］. Indian J Anim Res, 2014, 48(5): 464-468.
［3］Ruddell WSJ, Bone ES, Hill MJ, et al. Gastric-juice nitrite: a risk factor for cancer in the hypochlorhydric stomach［J］. Lancet, 1976, 308(7994): 1037-1039.
［4］Mirvish SS. Role of N-nitroso compounds (NOC) and N-nitrosation in etiology of gastric, esophageal, nasopharyngeal and bladder cancer and contribution to cancer of known exposures to NOC［J］. Cancer Lett, 1995, 93(1): 17-48.
［5］Brender JD, Olive JM, Felkner M, et al. Dietary nitrites and nitrates, nitrosatable drugs, and neural tube defects［J］. Epidemiology, 2004, 15(3): 330-336.
［6］Moriya M, Mitsumori K, Kato K, et al. Carcinogenicity of N-nitroso-ethylenethiourea in female mice［J］. Cancer Lett, 1979, 7(6): 339-342.
［7］Xi Y, Zhang JSh, Zang JF, et al. Stereological study on the synapse loss in visual cortex of mouse after prenatal alcohol exposure［J］. Acta Pharmaceutica Sinica, 2010, 45(6): 705-710.(in Chinese)
席艳, 张俊士, 臧建峰, 等. 孕期酒精暴露对子鼠视皮质突触数量影响的体视学研究［J］.药学学报, 2010, 45(6): 705-710.
［8］Inui N, Nishi Y, Taketomi M, et al. Transplacental action of sodium nitrite on embryonic cells of Syrian golden hamster［J］. Mutat Res, 1979, 66(2): 149-158.
［9］Brender JD, Werler MM, Kelley KE, et al. Nitrosatable drug exposure during early pregnancy and neural tube defects in offspring national birth defects prevention study［J］. Am J Epidemiol, 2011, 174(11): 1286-1295.
［10］Thiel G. Synapsin Ⅰ, synapsin Ⅱ, and synaptophysin: marker proteins of synaptic vesicles［J］. Brain Pathol, 1993, 3(1): 87-95.
［11］Hüls S, Hgen T, Vassallo N, et al. AMPA-receptor-mediated excitatory synaptic transmission is enhanced by iron-induced α-synuclein oligomers［J］. J Neurochem, 2011, 117(5): 868-878.
［12］Amtul Z, Atta-Ur-Rahman. Neural plasticity and memory: molecular mechanism［J］. Rev Neurosci, 2015, 26(3): 253-268.
［13］Galaburda A, Sanides F. Cytoarchitectonic organization of the human auditory cortex［J］.J Comp Neurol, 1980, 190(3): 597-610. 
［14］Chen R, Ruan GF, Fan ZhH. An investigation about acknowledgement of nitrite using in Jinan province, China［J］. Food and Nutrition in China, 2012, 18(3): 16-20.(in Chinese)
陈然, 阮光锋, 范志红. 济南市含亚硝酸盐餐饮配料使用和认知的调查［J］. 中国食物与营养, 2012, 18(3): 16-20. 
［15］Brender JD, Olive JM, Felkner M, et al. Dietary nitrites and nitrates, nitrosatable drugs, and neural tube defects［J］. Epidemiology, 2004, 15(3): 330-336.
［16］Olshan AF, Faustman EM. Nitrosatable drug exposure during pregnancy and adverse pregnancy outcome［J］. Int J Epidemiol, 1989, 18(4): 891-899. 
［17］Cedergren MI, Selbing AJ, Lfman O, et al. Chlorination byproducts and nitrate in drinking water and risk for congenital cardiac defects［J］. Environ Res, 2002, 89(2): 124-130. 
［18］Alder J, Xie ZP, Valtorta F, et al. Antibodies to synaptophysin interfere with transmitter secretion at neuromuscular synapses［J］. Neuron, 1992, 9(4): 759-768.
［19］Alder J, Lu B, Valtorta F, et al. Calciumdependent transmitter secretion reconstituted in Xenopus oocytes: requirement for synaptophysin［J］. Science, 1992, 257(5070): 657-661.
［20］Paula-Lima AC, Brito-Moreira J, Ferreira ST. Deregulation of excitatory neurotransmission underlying synapse failure in Alzheimer’s disease［J］. J Neurochem, 2013, 126(2): 191-202.
［21］Bennett MR, Farnell L, Gibson WG. Fiber pathway pathology, synapse loss and decline of cortical function in schizophrenia［J］. PLoS One, 2013, 8(4): e60518.
［22］Cheng F, Vivacqua G, Yu S. The role of α-synuclein in neurotransmission and synaptic plasticity［J］. J Chem Neuroanat, 2011, 42(4): 242-248.
［23］Krüger R, Müller T, Riess O. Involvement of alpha-synuclein in Parkinson’s disease and other neurodegenerative disorders［J］. J Neural Transm, 2000, 107(1): 31-40.
［24］Ginns EI, Mak SKK, Ko N, et al. Neuroinflammation and α-synuclein accumulation in response to glucocerebrosidase deficiency are accompanied by synaptic dysfunction［J］. Mol Genet Metab, 2014, 111(2): 152-162.
［25］Lavedan C. The synuclein family［J］. Genome Res, 1998, 8(9): 871-880.
［26］Zhang H, Maitta RW, Bhattacharyya PK, et al. γ-Synuclein is a promising new marker for staining reactive follicular dendritic cells, follicular dendritic cell sarcoma, Kaposi sarcoma, and benign and malignant vascular tumors［J］. Am J Surg Pathol, 2011, 35(12): 1857-1865.
［27］Buchman VL, Hunter HJ, Pin?n LG, et al. Persyn, a member of the synuclein family, has a distinct pattern of expression in the developing nervous system［J］. J Neurosci, 1998, 18(22): 9335-9341.
［28］Su JJ, Xie HJ, Zhao WW, et al. Study on γ-synuclein gene in patients with idiopathic Parkinson’s disease［J］. Chinese Journal of Medical Genetics, 2003, 20(5): 444-446. (in Chinese)
苏敬敬, 谢惠君, 赵武伟, 等. 散在帕金森病患者γ-synuclein基因的研究［J］. 中华医学遗传学杂志, 2003, 20(5): 444-446.

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