中华放射医学与防护杂志

黄志飞,周桂强,李姜怡,等.⁶⁰Co γ射线急性照射对小鼠情绪的影响及机制研究[J].中华放射医学与防护杂志,2025,45(12):1228-1236.Huang Zhifei,Zhou Guiqiang,Li Jiangyi,et al.Effects and their mechanisms of acute radiation with ⁶⁰Co γ-rays on the emotional state in exposed mice[J].Chin J Radiol Med Prot,2025,45(12):1228-1236

⁶⁰Co γ射线急性照射对小鼠情绪的影响及机制研究

Effects and their mechanisms of acute radiation with ⁶⁰Co γ-rays on the emotional state in exposed mice

投稿时间：2025-01-17

DOI：10.3760/cma.j.cn112271-20250117-00023

中文关键词: γ射线小鼠焦虑样情绪细胞焦亡星形胶质细胞

英文关键词:γ-ray Mice Anxiety-like behavior Pyroptosis Astrocyte

基金项目:陕西省重点研发计划项目(2024SF-YBXM-437)

作者	单位	E-mail
黄志飞	山东第二医科大学公共卫生学院, 潍坊 261053 空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
周桂强	空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
李姜怡	山东第二医科大学公共卫生学院, 潍坊 261053 空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
秦佟洲	空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
刘力源	空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
郑博	空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
王星	空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
王浩南	空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032
王春平	山东第二医科大学公共卫生学院, 潍坊 261053
丁桂荣	空军军医大学军事预防医学系辐射防护医学教研室特殊作业环境危害评估与防治教育部重点实验室, 西安 710032	dingzhao@fmmu.edu.cn

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中文摘要:

目的探讨⁶⁰Co γ射线单次全身照射对小鼠情绪的影响及其机制。方法采用随机数表法将48只6~8周龄雄性C57BL/6小鼠随机分为对照组、4 Gy组和6 Gy组，每组16只。照射组小鼠接受剂量率为1.91 Gy/min的⁶⁰Co γ射线4和6 Gy单次全身照射，照射后1、3、6周开展行为学检测，并在最后一次行为学实验结束后取材。采用开放旷场实验和高架十字迷宫实验评估小鼠的焦虑情绪；强迫游泳实验和悬尾实验评估小鼠的抑郁情绪；HE和Nissl染色观察小鼠海马脑区组织形态结构的变化；免疫组织化学染色检测小鼠海马脑区星形胶质细胞标志物GFAP和小胶质细胞标志物Iba-1表达水平；免疫荧光染色检测小鼠海马脑区细胞焦亡标志物GSDMD和Cleaved-caspase 1表达水平；Western blot检测小鼠海马脑区GFAP、Iba-1、GSDMD和Cleaved-caspase 1表达水平。结果与对照组相比，照射组小鼠在照后6周内未发生抑郁样行为改变，但在照射后3周出现了焦虑样行为，一直持续到照后6周未见恢复趋势，与4 Gy组相比，6 Gy组效应更为明显。病理学结果显示，照射组小鼠海马CA3脑区神经元出现深染、核固缩等变化，损伤神经元数量显著增多(F = 50.04，P＜0.05)，且6 Gy组改变更显著(t = 2.95，P＜0.05)。与对照组相比，6 Gy组小鼠海马CA3脑区GFAP阳性细胞数量显著增多(t = 3.27，P＜0.05)，两个照射组小鼠海马CA3脑区GSDMD和Cleaved-caspase 1阳性细胞数量均显著增多(F = 55.06、15.92，P＜0.05)，且6 Gy组有增加趋势。Western blot结果显示，与对照组相比，6 Gy组小鼠海马脑区GFAP蛋白表达水平显著升高(t = 2.92，P＜0.05)；两个照射组小鼠海马GSDMD和Cleaved-caspase 1蛋白水平均显著升高(F = 5.76、7.02，P＜0.05)。结论 4和6 Gy ⁶⁰Co γ射线全身急性照射可诱导小鼠产生焦虑样行为，且剂量越大效应越明显，与情绪调控相关的海马脑区细胞焦亡和星形胶质细胞活化可能参与了γ射线诱导情绪障碍的作用机制。

英文摘要:

Objective To explore the effects and their mechanisms of single whole-body exposure to ⁶⁰Co γ-rays on the emotional state in mice.Methods Totally 48 male C57BL/6 mice aged 6-8 weeks were randomly divided into a control, a 4 Gy irradiation group, and a 6 Gy irradiation group using the random number table method, with six mice in each group. Mice in the 4 and 6 Gy irradiation groups received a single whole-body irradiation with ⁶⁰Co γ-rays at a dose rate of 1.91 Gy/min, with doses of 4 Gy and 6 Gy, respectively. Then, behavioral experiments were conducted at 1, 3, and 6 weeks post-irradiation, followed by tissue sampling. The anxiety-like behavior of the mice was assessed using open field test (OFT) and elevated plus maze (EPM). Their depressive-like behavior was assessed using the forced swim test (FST) and tail suspension test (TST), and the morphological and structural changes of their hippocampus were examined using hematoxylin and eosin (HE) and Nissl staining. Furthermore, the expression levels of the astrocyte marker of glial fibrillary acidic protein (GFAP) and the microglial marker of Iba-1 in the hippocampus of the mice were detected using immunohistochemical (IHC) staining. The expression levels of the pyroptosis markers of Gasdermin D (GSDMD) and Cleaved-caspase1 in the hippocampus were examined using immunofluorescence (IF) staining, and the expression levels of GFAP, Iba-1, GSDMD, and Cleaved-caspase1 in the hippocampus were detected by Western blot analysis.Results Compared to the control group, the irradiation groups showed no depressive-like behavior at 6 weeks post-irradiation, but manifested anxiety-like behavior at 3 weeks post-irradiation. Furthermore, the anxiety-like behavior persisted until 6 weeks post-irradiation, with no signs of recovery. Compared to the 4 Gy irradiation group, such an effect was more pronounced in the 6 Gy irradiation group. Pathological result indicate that mice in both irradiation groups showed neuron changes in the hippocampal CA3 region, such as deep staining and nuclear condensation, and the number of damaged neurons increased significantly (F = 50.04, P < 0.05), with more significant changes observed in the 6 Gy irradiation group (t = 2.95, P < 0.05). The IHC and IF staining result showed that, compared to the control group, the 6 Gy irradiation group presented a significant increase in the number of GFAP-positive cells in the hippocampal CA3 region (t = 3.27, P < 0.05). Both irradiation groups displayed significant increases in the numbers of GSDMD and Cleaved-caspase 1 positive cells in the hippocampal CA3 region (F = 55.06, 15.92, P < 0.05), with an upward trend identified in the 6 Gy irradiation group. The Western blot analysis result indicated that, compared to the control group, the 6 Gy irradiation group showed a significant rise in the protein expression level of hippocampal GFAP (t = 2.92, P < 0.05). Both irradiation groups exhibited significantly elevated protein expression levels of hippocampal GSDMD and Cleaved-caspase 1 (F = 5.76, 7.02, P < 0.05).Conclusions Acute whole-body exposure to 4 Gy and 6 Gy ⁶⁰Co γ-rays can induce anxiety-like behavior in mice, with a certain extent of dose-effect relationship. The hippocampal pyroptosis and astrocyte activation associated with emotional regulation might be involved in the mechanism underlying γ-ray-induced emotional disturbance.

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