生酮饮食模型构建及行为学观察
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1. 南京医科大学附属儿童医院,南京 210008;2. 东南大学公共卫生学院,南京 210096;3. 南京医科大学基础医学院,南京 211166

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Establishment of a ketogenic diet model and behavioral evaluation in epileptic mice
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1. Children’s Hospital of Nanjing Medical University, Nanjing 210008, China; 2. School of Public Health,Southeast University, Nanjing 210096, China; 3. School of Basic Medical Sciences, Nanjing Medical University, Nanjing 211166, China

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    摘要:

    目的 本研究旨在建立颞叶癫痫小鼠模型,观察小鼠进食习惯,并在此基础上探究生酮饮食(ketogenic diet,KD)干预下小鼠体质量、毛发状态和血酮水平差异,进而探究 KD 对小鼠癫痫样发作及抑郁、焦虑相关行为学的影响。 方法 取 31 只 4 周龄 20 ~ 22 g 雄性 SPF 级 ICR 小鼠,取 11 只小鼠随机分配至常规饮食对照组(CON + ND 组,n= 5)和生酮饮食喂养组(CON + KD 组,n= 6);取 20 只小鼠通过腹腔注射匹罗卡品诱导癫痫持续状态(status epilepsy,SE),根据 Racine 分级评估每只小鼠癫痫发作级别,癫痫造模成功的小鼠随机平均分配至癫痫模型组(SE + ND 组)和癫痫模型生酮饮食干预组(SE + KD 组)。 实验周期设定为 28 d,CON + ND 组与 SE + ND 组接受标准饲料喂养,CON + KD 组与 SE + KD 组接受生酮饲料干预,构建规范的生酮饮食模型,监测小鼠体质量及尾静脉血酮值,采用视频监控系统记录癫痫发作次数,并通过旷场实验(open field test,OFT)、高架十字迷宫实验( elevated plus maze,EPM)及强迫游泳实验( forced swimming test,FST)比较不同组小鼠焦虑和抑郁样行为改变。 结果 (1)癫痫模型构建:20 只小鼠进行造模,4 只小鼠未达到 Racine 4 级,2 只小鼠死亡,14 只小鼠癫痫造模成功后随机平均分配至 SE + KD 组(n= 7)和 SE + ND 组(n= 7),SE + KD 组在生酮饮食喂养过程中 2 只小鼠死亡,SE + ND 组常规饲料喂养中无小鼠死亡;(2)生酮饮食模型构建:癫痫小鼠体质量低于正常小鼠,生酮饮食喂养和标准饲料喂养小鼠的体质量变化没有显著性差异(P>0. 05);CON + KD 组第 1、7、14、28 天血酮值均 ≥ 1. 0 mmol / L;CON + KD 组和 SE + KD 组小鼠在生酮饲料喂养第 2 天即出现毛发污染、排泄物附着及腹泻症状(n = 9),第 3 天腹泻严重的小鼠出现直肠脱垂(n =5);CON + ND 组和 SE + ND 组小鼠在常规饲料喂养期间保持毛发光泽且大便成形;(3)行为学实验:监测小鼠癫痫样行为发作频率,与 SE + ND 组比较,SE + KD 组发作次数减少;FST 中,与 CON + ND 组相比,CON +KD 组不动时间明显减少(P<0. 05)。 结论 规范化生酮饮食干预可有效维持治疗性酮症水平(血酮 ≥ 1. 0mmol / L),且对小鼠体质量未产生显著影响(P>0. 05)。 值得注意的是,生酮饮食显著抑制癫痫模型动物的异常放电活动(P<0. 05)。

    Abstract:

    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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蒋鑫蕊,李伟,涂福来,陈蕾,陈静,吴春风.生酮饮食模型构建及行为学观察[J].中国实验动物学报,2025,33(11):1610~1618.

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  • 收稿日期:2025-03-26
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  • 在线发布日期: 2026-01-08
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