电离辐射激活海马星形胶质细胞诱导小鼠认知功能障碍
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摘要
探讨电离辐射引起小鼠认知功能障碍的可能机制。24只雌性昆明小鼠随机分为对照组和辐照组,辐照组小鼠接受137Csγ射线单次全身辐照至吸收剂量4 Gy。35 d后采用Y迷宫实验检测小鼠认知功能;免疫组织化学法检测小鼠海马区星形胶质细胞(Astrocytes,Ast)标记物胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)阳性细胞数的表达变化;Western blot方法检测海马GFAP蛋白的表达变化;荧光定量PCR检测炎性因子白细胞介素IL-1α、IL-1β、IL-6 mRNA和海马GFAP的表达情况。结果表明:与对照组相比,辐照组小鼠在Y迷宫实验中自发交替率明显降低(78.40±4.16 vs. 67.53±3.08,p<0.05);海马区GFAP阳性细胞数明显增加(133.4±9.6 vs. 280.3±32.7,p<0.01);GFAP蛋白及mRNA表达均显著上调(1.000±0.036vs. 1.300±0.102,p<0.05;1.000±0.031 vs. 1.203±0.078,p<0.05);IL-1α、IL-1β和IL-6 mRNA的表达均明显上调(1.000±0.133 vs. 1.677±0.164,p<0.01;1.000±0.132 vs. 1.488±0.105,p<0.05;1.000±0.218 vs.2.181±0.188,p<0.01)。研究提示,电离辐射可能是通过激活海马Ast诱导炎症因子释放,从而引起认知功能障碍。
In order to explore the possible mechanisms underlying cognitive dysfunction induced by ionizing radiation in mice, twenty-four female Kunming mice were randomly divided into a control group and irradiation group(IR). The IR group mice were irradiated with137 Cs γ-rays at an absorbed dose of 4 Gy. After 35 d, the Y-maze test was used to evaluate the cognitive function of the mice. The expression of the astrocyte marker, glial fibrillary acidic protein(GFAP), was detected using immunohistochemical staining. The expression of GFAP protein in the hippocampus was assayed using western blot. The expression of GFAP, interleukin(IL)-1α, IL-1β, and IL-6 mRNA in the hippocampus was detected using reverse transcription polymerase chain reaction. The results showed that compared with the control group, the spontaneous alternation rate of the IR mice was significantly reduced in the Ymaze test(78.40 ± 4.16 vs. 67.53 ± 3.08, p < 0.05); and the number of GFAP-positive cells in the hippocampus was significantly higher(133.4 ± 9.6 vs. 280.3 ± 32.7, p < 0.01); GFAP protein levels(1.000 ± 0.036 vs. 1.300 ± 0.102, p<0.05) and GFAP mRNA expression(1.000 ± 0.0307 vs. 1.203 ± 0.078, p < 0.05) were significantly upregulated;and the expression of inflammatory factors IL-1α, IL-1β, and IL-6 mRNA was upregulated(1.000 ± 0.133 vs 1.677 ±0.164, p < 0.01; 1.000 ± 0.132 vs. 1.488 ± 0.105, p < 0.05; 1.000 ± 0.218 vs 2.181 ± 0.188, p < 0.01). It is suggested that ionizing radiation induces cognitive dysfunction by activating astrocytes in the hippocampus.
In order to explore the possible mechanisms underlying cognitive dysfunction induced by ionizing radiation in mice, twenty-four female Kunming mice were randomly divided into a control group and irradiation group(IR). The IR group mice were irradiated with137 Cs γ-rays at an absorbed dose of 4 Gy. After 35 d, the Y-maze test was used to evaluate the cognitive function of the mice. The expression of the astrocyte marker, glial fibrillary acidic protein(GFAP), was detected using immunohistochemical staining. The expression of GFAP protein in the hippocampus was assayed using western blot. The expression of GFAP, interleukin(IL)-1α, IL-1β, and IL-6 mRNA in the hippocampus was detected using reverse transcription polymerase chain reaction. The results showed that compared with the control group, the spontaneous alternation rate of the IR mice was significantly reduced in the Ymaze test(78.40 ± 4.16 vs. 67.53 ± 3.08, p < 0.05); and the number of GFAP-positive cells in the hippocampus was significantly higher(133.4 ± 9.6 vs. 280.3 ± 32.7, p < 0.01); GFAP protein levels(1.000 ± 0.036 vs. 1.300 ± 0.102, p<0.05) and GFAP mRNA expression(1.000 ± 0.0307 vs. 1.203 ± 0.078, p < 0.05) were significantly upregulated;and the expression of inflammatory factors IL-1α, IL-1β, and IL-6 mRNA was upregulated(1.000 ± 0.133 vs 1.677 ±0.164, p < 0.01; 1.000 ± 0.132 vs. 1.488 ± 0.105, p < 0.05; 1.000 ± 0.218 vs 2.181 ± 0.188, p < 0.01). It is suggested that ionizing radiation induces cognitive dysfunction by activating astrocytes in the hippocampus.
引文
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16 Alexander T C,Butcher H,Krager K,et al.Behavioral effects of focal irradiation in a juvenile murine model[J].Radiation Research,2018,189(6):605-617.DOI:10.1667/RR14847.1.
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19 Colangelo A M,Cirillo G,Lavitrano M L,et al.Targeting reactive astrogliosis by novel biotechnological strategies[J].Biotechnology Advances,2012,30(1):261-271.DOI:10.1016/j.biotechadv.2011.06.016.
20谈丹丹,洪道俊,徐仁伵,等.神经胶质细胞和神经退行性疾病[J].中国老年学杂志,2013,33(2):464-466.DOI:10.3969/j.issn.1005-9202.2013.02.114.TAN Dandan,HONG Daojun,XU Rensi,et al.Glial cells and neurodegenerative diseases[J].Chinese Journal of Gerontology,2013,33(2):464-466.DOI:10.3969/j.issn.1005-9202.2013.02.114.
21 Franciosi S,Choi H B,Kim S U,et al.IL-8 enhancement of amyloid-beta(Abeta 1-42)-induced expression and production of pro-inflammatory cytokines and COX-2 in cultured human microglia[J].Journal of Neuroimmunology,2005,159(1-2):66-74.DOI:10.1016/j.jneuroim.2004.10.006.
22 Mattson M P.Pathways towards and away from Alzheimer's disease[J].Nature,2004,430(7000):631-639.DOI:10.1038/nature02621.
23 Garber C,Vasek M J,Vollmer L L,et al.Astrocytes decrease adult neurogenesis during virus-induced memory dysfunction via IL-1[J].Nature Immunology,2018,19(2):151-161.DOI:10.1038/s41590-017-0021-y.
2 Tang Y,Luo D,Rong X,et al.Psychological disorders,cognitive dysfunction and quality of life in nasopharyngeal carcinoma patients with radiationinduced brain injury[J].PLoS One,2012,7(6):e36529.DOI:10.1371/journal.pone.0036529.
3 Eichenbaum H.Hippocampus:cognitive processes and neural representations that underlie declarative memory[J].Neuron,2004,44(1):109-120.
4 Zou Y,Corniola R,Leu D,et al.Extracellular superoxide dismutase is important for hippocampal neurogenesis and preservation of cognitive functions after irradiation[J].Proceeding of the National Academy of Sciences of the United States of America,2012,109(52):21522-21527.DOI:10.1073/pnas.1216913110.
5 Lee W H,Sonntag W E,Lee Y W.Aging attenuates radiation-induced expression of pro-inflammatory mediators in rat brain[J].Neuroscience Letters,2010,476(2):89-93.DOI:10.1016/j.neulet.2010.04.009.
6 Shastri A,Bonifati D M,Kishore U.Innate immunity and neuroinflammation[J].Mediators of Inflammation,2013,2013:342931.DOI:10.1155/2013/342931.
7田乃元,吴旭,卢岩.星形胶质细胞与认知功能的研究进展[J].解剖科学进展,2014(3):283-287.DOI:10.16695/j.cnki.1006-2947.2014.03.001.TIAN Naiyuan,WU Xu,LU Yan.Progress of the astrocytes and cognitive function[J].Progress of Anatomical Sciences,2014(3):283-287.DOI:10.16695/j.cnki.1006-2947.2014.03.001.
8 Garwood C J,Pooler A M,Atherton J,et al.Astrocytes are important mediators of Aβ-induced neurotoxicity and tau phosphorylation in primary culture[J].Cell Death and Disease,2011,2:e167.DOI:10.1038/cddis.2011.50.
9胡雅梦,刘琅嬛,刘宝婵,等.黄芪甲苷对肝细胞的辐射防护作用[J].辐射研究与辐射工艺学报,2015,33(6):060202.DOI:10.11889/j.1000-3436.2015.rrj.33.060202.HU Yameng,LIU Langhuan,LIU Baochan,et al.Radioprotective effects of astragaloside IV on the liver cell[J].Journal of Radiation Research and Radiation Processing,2015,33(6):060202.DOI:10.11889/j.1000-3436.2015.rrj.33.060202.
10乃爱桃,王贞,曹文宇,等.丰富环境对辐射诱导小鼠认知障碍的保护作用及可能机制[J].中华放射医学与防护杂志,2018,38(6):401-406.DOI:10.3760/cma.j.issn.0254-5098.2018.06.001.NAI Aitao,WANG Zhen,CAO Wenyu,et al.The protective effect of environmental enrichment on radiation induced cognitive dysfunction and underlying mechanism[J].Chinese Journal of Radiological Medicine and Protection,2018,38(6):401-406.DOI:10.3760/cma.j.issn.0254-5098.2018.06.001.
11 Cao W,Duan J,Wang X,et al.Early enriched environment induces an increased conversion of proBDNF to BDNF in the adult rat's hippocampus[J].Behavioural Brain Research,2014,265:76-83.DOI:10.1016/j.bbr.2014.02.022.
12田丹,王彦,王芹,等.LGK-974对人肝癌细胞系HepG2的辐射增敏作用[J].辐射研究与辐射工艺学报,2016,34(6):060203.DOI:10.11889/j.1000-3436.2016.rrj.34.060203.TIAN Dan,WANG Yan,WANG Qin,et al.Effects of LGK-974 in enhancing the radiosensitivity of HepG2hepatoma cells[J].Journal of Radiation Research and Radiation Processing,2016,34(6):060203.DOI:10.11889/j.1000-3436.2016.rrj.34.060203.
13李曙芳,黄立群,王永丽,等.重度骨髓型急性放射病小鼠模型的建立[J].毒理学杂志,2015,29(6):450-453.DOI:10.16421/j.cnki.1002-3127.2015.06.014.LI Shufang,HUANG Liqun,WANG Yongli,et al.Establishment of a mouse model of severe myeloid acute radiation disease[J].Journal of Toxicology,2015,29(6):450-453.DOI:10.16421/j.cnki.1002-3127.2015.06.014.
14王利,翟瑞仁,逄朝霞,等.BALB/c小鼠重度骨髓型急性放射病模型的建立[J].肿瘤研究与临床,2012,24(11):725-727.DOI:10.3760/cma.j.issn.1006-9801.2012.11.002.WANG Li,ZHAI Ruiren,PANG Zhaoxia,et al.Establishment of an animal model of severe bone marrow type acute radiation sickness in the BALB/c mice[J].Journal of Toxicology,2012,24(11):725-727.DOI:10.3760/cma.j.issn.1006-9801.2012.11.002.
15乃爱桃,王贞,唐丽云,等.伽马射线辐照对小鼠认知功能及神经元的影响[J].辐射研究与辐射工艺学报,2018,36(5):050202.DOI:10.11889/j.1000-3436.2018.rrj.36.050202.NAI Aitao,WANG Zhen,TANG Liyun,et al.Effects of gamma-ray irradiation on cognitive function and neuronal death in mice[J].Journal of Radiation Research and Radiation Processing,2018,36(5):050202.DOI:10.11889/j.1000-3436.2018.rrj.36.050202.
16 Alexander T C,Butcher H,Krager K,et al.Behavioral effects of focal irradiation in a juvenile murine model[J].Radiation Research,2018,189(6):605-617.DOI:10.1667/RR14847.1.
17 Achanta P,Fuss M,Martinez J J.Ionizing radiation impairs the formation of trace fear memories and reduces hippocampal neurogenesis[J].Behavioral Neuroscience,2009,123(5):1036-1045.DOI:10.1037/a0016870.
18 Voskuhl R R,Peterson R S,Song B,et al.Reactive astrocytes form scar-like perivascular barriers to leukocytes during adaptive immune inflammation of the CNS[J].Journal of Neuroscience,2009,29(37):11511-11522.DOI:10.1523/JNEUROSCI.1514-09.2009.
19 Colangelo A M,Cirillo G,Lavitrano M L,et al.Targeting reactive astrogliosis by novel biotechnological strategies[J].Biotechnology Advances,2012,30(1):261-271.DOI:10.1016/j.biotechadv.2011.06.016.
20谈丹丹,洪道俊,徐仁伵,等.神经胶质细胞和神经退行性疾病[J].中国老年学杂志,2013,33(2):464-466.DOI:10.3969/j.issn.1005-9202.2013.02.114.TAN Dandan,HONG Daojun,XU Rensi,et al.Glial cells and neurodegenerative diseases[J].Chinese Journal of Gerontology,2013,33(2):464-466.DOI:10.3969/j.issn.1005-9202.2013.02.114.
21 Franciosi S,Choi H B,Kim S U,et al.IL-8 enhancement of amyloid-beta(Abeta 1-42)-induced expression and production of pro-inflammatory cytokines and COX-2 in cultured human microglia[J].Journal of Neuroimmunology,2005,159(1-2):66-74.DOI:10.1016/j.jneuroim.2004.10.006.
22 Mattson M P.Pathways towards and away from Alzheimer's disease[J].Nature,2004,430(7000):631-639.DOI:10.1038/nature02621.
23 Garber C,Vasek M J,Vollmer L L,et al.Astrocytes decrease adult neurogenesis during virus-induced memory dysfunction via IL-1[J].Nature Immunology,2018,19(2):151-161.DOI:10.1038/s41590-017-0021-y.