电离辐射致认知功能损伤大鼠模型的研究
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(中国辐射防护研究院,药物毒理与放射损伤药物山西省重点实验室, 太原 030006)

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R-33


Establishment of a rat model of cognitive impairment induced by ionizing radiation
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(Key Laboratory of Drug Toxicology and Drug for Radiation Injury, China Institute for Radiation Protection,Taiyuan 030006,China)

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    摘要:

    目的 对大鼠头部进行不同剂量照射,选取合适剂量建立电离辐射致认知功能损伤动物模型,为研究电离辐射致认知功能损伤发生机制?辐射防护剂研制等提供研究基础?方法 将SPF 级雄性大鼠20 只随机分为对照组(C 组)?10 Gy 照射组(10 Gy IR 组)?20 Gy 照射组(20 Gy IR 组)和30 Gy 照射组(30 Gy IR 组),每组各5只,C 组不作处理,其余3 组分别利用10 Gy?20 Gy 和30 Gy 电子线对不同组别SD 大鼠头部进行单次照射,照射后30 d 计算动物死亡率,利用Morris 水迷宫对大鼠空间记忆能力进行检测,利用化学比色法检测大鼠脑皮质匀浆液中还原型谷胱甘肽( glutathione, GSH)?丙二醛( malondialdehyde, MDA), 酶联免疫吸附法( enzyme-linked immunoSorbent assay, ELISA)检测8 羟基脱氧鸟苷(8-hydroxydeoxyguanosine, 8-OHdG)含量,对经HE 染色的大脑病理切片进行检测观察?结果 30 Gy IR 组动物出现背毛不整齐,流涎等症状,头颈部发生严重的脱毛现象?照后30 d 时,C 组未见动物死亡,10 Gy IR 组和20 Gy IR 组动物死亡率为20%,30 Gy IR 组动物死亡率为40%?Morris水迷宫定位航行实验结果显示30 Gy IR 组在1~5 d 的训练期中,潜伏期时间均高于其余三组且差异具有统计学意义( P <0. 05)?空间搜索实验显示30 Gy IR 组较其余各组原平台象限停留时间显著缩短( P <0. 05);穿越平台次数显著减少( P <0. 05);10 Gy IR 组?20 Gy IR 组?30 Gy IR 组原平台象限游泳距离较C 组均显著缩短( P <0. 05);氧化应激相关指标检测结果显示30 Gy IR 组与C 组相比GSH?8-OHdG?MDA 浓度差异有统计学意义( P <0. 05),20 Gy IR 组与C 组MDA 浓度差异有统计学意义( P <0. 05);HE 染色切片发现20 Gy IR 组和30 Gy IR 组组织结构破坏较10 Gy IR 组更加严重,出现组织细胞凋亡?坏死等显著的病理改变?结论 本次实验结果提示对头部进行30 Gy 单次照射可以成功建立大鼠认知功能障碍动物模型?

    Abstract:

    Objective To investigate the effects of exposure of the head of rats to different doses of ionizing radiation, to establish an appropriate animal model of cognitive function damage caused by such radiation, and to provide a research basis for studying the mechanism of cognitive function damage due to ionizing radiation and the development of radiation-protective agents. Methods Twenty male SPF rats were randomly and equally divided into a control group (C group), a 10 Gy irradiation group (10 Gy IR group), a 20 Gy irradiation group (20 Gy IR group), and a 30 Gy irradiation group (30 Gy IR group). The group C was left untreated, while the other three groups were irradiated with 10,20, and 30 Gy electron beams, focused on the heads of the SD rats. The mortality of the animals was calculated using survival rates at 30 days after the irradiation and spatial memory was assessed using the Morris water maze. The concentrations of glutathione (GSH) and malondialdehyde (MDA) in rat brain cortical homogenate were determined by chemical colorimetry. An enzyme-linked immunosorbent assay (ELISA) was used to detect 8-hydroxydeoxyguanosine (8-OHdG). Brain pathological sections were analyzed by HE staining. Results The rats in the 30 Gy IR group exhibited irregular back hair, excessive salivation, and other symptoms, along with severe hair loss on the head and neck. At 30 days after the irradiation, no animal died in group C. The mortality rates of both the 10 and 20 Gy IR groups were 20%, while that of the 30 Gy IR group was 40%. The results regarding positioning and navigation in the Morris water maze experiment showed that the latency of the 30 Gy IR group in the 1-5-day training period was significantly higher than in the other three groups ( P <0. 05). The spatial search experiment showed that the residence time in the original platform quadrants of the 30 Gy IR group was significantly shorter than in the other groups ( P <0. 05); the number of platform crosses in this group was also significantly reduced ( P <0. 05). The swimming distances in the original platform for the 10, 20, and 30 Gy IR groups were significantly shorter than for the group C ( P < 0. 05). Moreover, the results of oxidative stress-related indicators showed that the concentrations of GSH, 8-OHdG, and MDA in the 30 Gy IR group were significantly different from those in group C ( P <0. 05). The difference of MDA concentration between the 20 Gy IR group and group C was statistically significant ( P <0. 05). Pathological examination also showed that the tissue damage of the 20 and 30 Gy IR groups was more severe than that of the 10 Gy IR group, and there were significant pathological changes such as cell apoptosis and tissue necrosis. Conclusions This experiment shows that a rat model of cognitive dysfunction can be created by a single 30 Gy electron beam irradiation to the head.

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刘梦雅,袁慧,高洁,王永丽,尹晶晶,王新钢,刘欢,李建国,秦秀军.电离辐射致认知功能损伤大鼠模型的研究[J].中国比较医学杂志,2019,29(2):72~77.

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  • 收稿日期:2018-08-20
  • 在线发布日期: 2019-03-14
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