模拟急性辐射综合征的Beclin1基因敲除小鼠模型

Q1 Health Professions

Radiation Medicine and Protection Pub Date : 2025-08-01 DOI:10.1016/j.radmp.2025.06.004

Wen Wei , Lei Li , Zhenzhen Liu , Xueqin Gao , Xueqing Wang , Jianrong Wang , Yixuan Fang , Na Yuan

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引用次数: 0

摘要

目的探讨Beclin-1 （Becn1）蛋白在辐射暴露小鼠中的作用。方法采用sa基因靶向策略制备Becn1-floxed小鼠，与Ubc-iCre小鼠杂交，制备条件基因敲除小鼠（又称Becn1f/f；Ubc-iCre小鼠）。然后用他莫昔芬（TMXF）诱导产生Becn1敲除小鼠。Becn1基因敲除小鼠和受致命辐射的小鼠在各自的中位生存时间前一天被安乐死。取其心、肝、脾、肺、肾、肠等脏器组织进行检查。此外，通过遗传、组织学和功能分析比较Becn1敲除小鼠和致死辐照小鼠。结果全身Becn1基因敲除的小鼠与暴露于致死剂量γ射线照射的小鼠表现出相似的表型，包括生存时间缩短（KO组中位生存时间为8-9天，而照射组中位生存时间为8-11天）、各组织器官形态和病理改变、造血系统破坏和DNA损伤。苏木精和伊红（H&；E）染色切片显示Becn1基因敲除和致死照射小鼠的类似病理变化，如脾脏结构破坏，白色髓减少，胸腺滤泡变性，股骨内有核细胞明显减少，肠绒毛广泛脱落。这些小鼠的造血干细胞和祖细胞（HSPCs）增殖和分化能力受损，出现类似的DNA损伤指标，如骨髓、心脏、脾脏和胸腺中活性氧（ROS）水平升高和γ-H2AX表达增加。值得注意的是，小鼠的Becn1蛋白在辐射照射后6小时内迅速降解。结论成年小鼠Becn1的全身双等位基因缺失模拟了致死辐射的效应，表明Becn1是一种高辐射敏感蛋白。

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A Beclin1 knockout mouse model mimicking acute radiation syndrome

Objective

To determine the role of the Beclin-1 (Becn1) protein in radiation-exposed mice.

Methods

A gene targeting strategy was employed to generate Becn1-floxed mice, which were then crossed with Ubc-iCre mice to create preconditional gene knockout mice (also referred to as Becn1^f/f;Ubc-iCre mice). Then, tamoxifen (TMXF) induction was used to generate Becn1 knockout mice. Both Becn1 knockout and lethally irradiated mice were euthanized a day before their respective median survival time. Their organs and tissues including the heart, liver, spleen, lung, kidney, and intestine were collected for examination. Furthermore, the Becn1 knockout and lethally irradiated mice were compared through genetic, histological, and functional analyses.

Results

Mice subjected to systemic Becn1 gene knockout and those exposed to a lethal dose of γ-ray irradiation exhibited similar phenotypes, including reduced survival time (median survival: 8–9 d for KO vs. 8–11 d for irradiated), morphological and pathological changes in various tissues and organs, hematopoietic system disruptions, and DNA damage. Hematoxylin and eosin (H&E)-stained sections showed analogous pathological changes in both the Becn1 knockout and lethally irradiated mice, such as the disrupted splenic architecture with decreased white pulp, degenerating thymic follicles, significantly reduced nucleated cells within the femur, and extensively denuding intestinal villi. These mice demonstrated impaired proliferation and differentiation capacities of hematopoietic stem and progenitor cells (HSPCs), presenting similar DNA damage indicators, such as heightened reactive oxygen species (ROS) levels and increased γ-H2AX expression in the bone marrow, heart, spleen, and thymus. Notably, the Becn1 protein in the mice underwent rapid degradation within 6 h after radiation exposure.