大鼠中敲除 SOD1 导致肌萎缩侧索硬化症(ALS)表型
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1. 中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,呼吸和共病全国重点实验室,国家卫生健康委员会比较医学重点实验室,教育部病原体感染防控教育部重点实验室,国家动物模型技术创新中心,国家人类疾病动物模型资源库,北京 100021;2. 中国医学科学院医学实验动物研究所,灵长类研究中心,北京 100021;3. 细胞生态海河实验室,天津 300301

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Establishment of a ketogenic diet model and behavioral evaluation in epileptic mice
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1.State Key Laboratory of Respiratory Health and Multimorbidity, NHC Key Laboratory of Human Disease Comparative Medicine, Key Laboratory of Pathogen Infection Prevention and Control Ministry of Education,Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences & Comparative Medicine Center of Peking Union Medical College, Beijing 100021, China; 2. Medical Primate Research Center,Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Beijing 100021, China;3. Haihe Laboratory of Cell Ecosystem, Tianjin 300301, China

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    摘要:

    目的 建立超氧化物歧化酶 1 ( SOD1) 基因敲除大鼠,解析其行为学表型。 方法 采用CRISPR/ Cas9 技术构建 SOD1 敲除型大鼠,通过 PCR 及 Sanger 测序进行基因型鉴定,Western Blot 验证 SOD1蛋白表达情况。 与野生型大鼠交配,评估基因型遗传稳定性。 动态监测体质量变化、生存率,系统评估组织病变、行为学异常、脊髓运动神经元数量变化及炎症变化情况。 结果 基因型鉴定确认 SOD1 敲除型大鼠模型制备成功。 SOD1 基因缺失影响大鼠发育及体质量。 行为学显示敲除型大鼠表现出渐进性的运动共济失调,最终导致后肢瘫痪,伴随有脊髓 L4 区运动神经元丧失,小胶质细胞显著增多、腓肠肌肌细胞数量与横截面积减少等肌萎缩侧索硬化症(ALS)疾病特征。 结论 本研究成功构建 SOD1 敲除型大鼠模型,该模型能重现ALS 疾病的一些核心病理特征,为深入研究 SOD1 基因在 ALS 中的功能提供了重要动物模型。

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    Objective To establish a mouse model of temporal lobe epilepsy and to investigate the effects of the ketogenic diet (KD). Specifically, we aimed to compare the differences in body mass, fur condition, and blood ketone levels between mice on the KD and those on a regular diet; and to further explore the impact of KD on seizure susceptibility, as well as depression- and anxiety-like behaviors. Methods Thirty-one 4-weeks-old male SPF-grade ICR mice ( 20 ~ 22 g) underwent acclimatization for 1 week. Eleven mice were assigned randomly to control conventional diet (CON + ND, n= 5) and KD (CON + KD, n= 6) groups. The remaining 20 mice were subjected to pilocarpine-induced status epilepticus (SE). The SE model was validated using Racine scale scoring (≥ stage 4),and successfully modeled mice were allocated randomly to epileptic standard diet (SE + ND) and epileptic ketogenic diet group (SE + KD) groups. Seizure frequency was recorded via video monitoring, and anxiety- and depression-like behaviors were assessed using open field, elevated plus maze, and forced swimming tests ( FST). Results ( 1) Establishment of epilepsy models: of the 20 mice subjected to modeling, four failed to reach Racine stage 4 and two died. The remaining 14 successfully modeled mice were allocated randomly to SE + KD (n= 7) and SE + ND (n=7) groups. During intervention, two mice died in the SE + KD group, while no mortality occurred in the SE + ND group. (2)Establishment of the ketogenic diet model: epileptic mice had lower baseline body mass than controls, but there was no significant difference in weight change between the dietary interventions ( P>0. 05). Blood ketone levels in the CON + KD group were consistently ≥ 1. 0 mmol / L at days 1, 7, 14, and 28. By day 2 of ketogenic feeding, nine mice developed fur soiling with fecal matter and diarrhea, progressing to rectal prolapse in five severe cases by day 3. CON + ND and SE + ND mice retained glossy coats and formed stools throughout. (3) Behavioral tests: SE + KD mice had fewer seizures compared with SE + ND mice. In the FST, CON + KD exhibited shorter immobility times than CON + ND mice. Conclusions Standardized KD intervention effectively maintained therapeutic ketosis (blood ketones ≥ 1. 0 mmol / L) without significant weight impact. Notably, a KD suppressed epileptic discharges and ameliorated anxiety- and depression-like behaviors.

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李新月,潘烁,张磊,贺佳跃,刘宁,张旭,高珊,齐晓龙,马元武.大鼠中敲除 SOD1 导致肌萎缩侧索硬化症(ALS)表型[J].中国实验动物学报,2025,33(11):1598~1609.

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  • 收稿日期:2025-08-13
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  • 在线发布日期: 2026-01-08
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