中华放射医学与防护杂志

吴卓君,黄志飞,李姜怡,等.不同剂量电离辐射对小鼠肠道NADPH氧化酶表达的影响[J].中华放射医学与防护杂志,2025,45(5):399-407.Wu Zhuojun,Huang Zhifei,Li Jiangyi,et al.Impacts of ionizing radiation dose on NADPH oxidase expression in mouse intestine[J].Chin J Radiol Med Prot,2025,45(5):399-407

不同剂量电离辐射对小鼠肠道NADPH氧化酶表达的影响

Impacts of ionizing radiation dose on NADPH oxidase expression in mouse intestine

投稿时间：2024-10-30

DOI：10.3760/cma.j.cn112271-20241030-00417

中文关键词: 电离辐射双重氧化酶2(DUOX2) 小肠烟酰胺腺嘌呤二核苷酸磷酸(NADPH) 氧化酶

英文关键词:Ionizing radiation Dual oxidase 2 (DUOX2) Intestine Nicotinamide adenine dinucleotide phosphate (NADPH)

基金项目:

作者	单位	E-mail
吴卓君	空军军医大学, 西京消化病医院, 国家消化系统疾病临床医学研究中心和消化系肿瘤整合防治全国重点实验室, 西安 710032
黄志飞	空军军医大学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	空军军医大学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
郭玲	空军军医大学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
丁桂荣	空军军医大学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:

目的探讨不同剂量电离辐射对小鼠肠道烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶和活性氧水平变化的影响。方法将6~8周龄的C57BL/6小鼠随机数表法分为0 Gy组、0.1 Gy组、0.2 Gy组和0.5 Gy照射组,每组10只。使用⁶⁰Co γ射线对小鼠进行单次全身辐射,剂量分别为0、0.1、0.2和0.5 Gy,吸收剂量率为13 mGy/min。照射后20周收集小鼠空肠,回肠及结肠组织,采用免疫组织化学、蛋白免疫印记和反转录聚合酶链反应检测NADPH氧化酶mRNA和蛋白表达情况,并测定肠组织中过氧化氢(H₂O₂)水平以及双重氧化酶2 (dual oxidase 2,DUOX2) 的启动器NF-κB (Nuclear Factor-kappa B) 的表达情况。结果与0 Gy组小鼠相比,0.5 Gy组小鼠空肠中绒毛长度变短,回肠绒毛融合,结肠中隐窝间隙变大,差异有统计学意义(t=2.48,P < 0.05),其余剂量照射组与0 Gy组相比差异无统计学意义(P >0.05)。与0 Gy组相比,0.1 Gy组小鼠空肠、回肠、结肠中H₂O₂的表达升高 (t=4.12,3.12,3.12,P < 0.05);0.5 Gy组与0 Gy相比,空肠和结肠中H₂O₂的表达升高近1倍 (t=8.67,8.69,P < 0.05)。辐照后20周,与0 Gy组相比,DUOX2蛋白在空肠、回肠、结肠中表达水平明显升高 (t=3.03,10.29,2.74,P < 0.05) 。与0 Gy组相比,0.1 Gy组小鼠空肠、回肠和结肠组织 DUOX2 mRNA水平显著升高,差异具有统计学意义 (t=12.75,4.12,11.14,P < 0.05); 与0 Gy组相比,空肠中0.2 Gy组NOX4 mRNA相对表达量升高 (t=4.54,P < 0.05);与0 Gy组相比,回肠中0.1 Gy组NOX4 mRNA升高 (t=4.13,P < 0.05)。与0 Gy组相比,空肠、回肠、结肠中0.1 Gy组DUOX2的启动器核因子κB(nuclear factor kappa-B,NF-κB)的表达有上调趋势,差异均有统计学意义 (t=8.73,8.18,7.02,P < 0.05)。结论小剂量射线照射对小肠具有远后效应,具体表现在0.5 Gy照射可导致小鼠空肠、回肠和结肠形态学结构轻微改变, 0.1 Gy照射可促进肠道NADPH氧化酶DUOX2表达增加。

英文摘要:

Objective To investigate the effects of different doses of ionizing radiation on the changes in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and reactive oxygen species (ROS) levels in the intestines of mice. Methods C57BL/6 mice aged 6-8 weeks were randomly assigned to four groups (0 Gy, 0.1 Gy, 0.2 Gy, and 0.5 Gy; n=10/group) and subjected to single whole-body irradiation using a ⁶⁰Co γ-ray source at a dose rate of 13 mGy/min. At 20 weeks post-irradiation, jejunal, ileal, and colonic tissues were collected. Immunohistochemistry, Western blotting, and reverse transcription-polymerase chain reaction (RT-PCR) were employed to assess mRNA and protein expression of NADPH oxidase components. Hydrogen peroxide (H₂O₂) levels and the expression of Nuclear Factor-kappa B (NF-κB), a transcriptional regulator of Dual Oxidase 2 (DUOX2), were also measured. Results Compared with the 0 Gy group, mice in the 0.5 Gy group exhibited shortened villus length in the jejunum, villus fusion in the ileum, and increased crypt spacing in the colon, with statistically significant differences (t=2.48, P < 0.05). No significant differences were observed in other dose groups compared to the 0 Gy group (P > 0.05).The expression of H₂O₂ in the jejunum, ileum, and colon of the 0.1 Gy group was significantly elevated compared to the 0 Gy group (t=4.12, 3.12, 3.12; P < 0.05). In the 0.5 Gy group, H₂O₂ expression in the jejunum and colon increased nearly twofold relative to the 0 Gy group (t=8.67, 8.69; P < 0.05).At 20 weeks post-irradiation, DUOX2 protein expression levels in the jejunum, ileum, and colon were markedly higher in irradiated mice than in the 0 Gy group (t=3.03, 10.29, 2.74; P < 0.05). DUOX2 mRNA levels in the jejunum, ileum, and colon of the 0.1 Gy group were significantly upregulated compared to the 0 Gy group (t=12.75, 4.12, 11.14; P < 0.05). Additionally, NOX4 mRNA expression increased in the jejunum of the 0.2 Gy group (t=4.54, P < 0.05) and in the ileum of the 0.1 Gy group (t=4.13, P < 0.05).The nuclear factor kappa-B (NF-κB), a transcriptional regulator of DUOX2, showed an upward trend in expression in the jejunum, ileum, and colon of the 0.1 Gy group compared to the 0 Gy group, with statistically significant differences (t=8.73, 8.18, 7.02; P < 0.05). Conclusion Low-dose radiation induces long-term effects on the intestinal tract. Specifically, 0.5 Gy irradiation causes mild morphological alterations in the jejunum, ileum, and colon, while 0.1 Gy irradiation promotes the upregulation of DUOX2, a NADPH oxidase, in intestinal tissues.

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