小鼠酒精性心肌病模型饲料和模型复制方法

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专题

不同动物酒精性心肌病模型饲料
和模型复制方法

小鼠酒精性心肌病(ACM)
模型饲料和模型复制方法
Alcoholic Cardiomyopathy Murine Model (ACM mouse Model) and Diet

在“酒精性心肌病模型饲料和造模方法”中，我们已经介绍了ACM（alcoholic cardiomyopathy)建立的总体情况，现在我们详细介绍使用小鼠建立ACM。如果想了解大鼠如何建立ACM，请见：大鼠酒精性心肌病模型饲料和模型复制方法。

酒精性心肌病模型需要给予酒精的时间较长，严格地论证造模方法非常关键，马虎不得。因此，在选择动物和模型饲料方面，建议研究者在阅读后与南通特洛菲饲料科技有限公司进行技术交流，尽可能争取获得满意的模型。

实验技术

www.trophic.cn

2014年1月21日出版

小鼠酒精性心肌病模型复制方法

Alcoholic Cardiomyopathy Murine Model (ACM mouse Model) and Diet

南通特洛菲饲料科技有限公司技术部

【摘要】本文介绍了应用小鼠进行酒精性心肌病（mouse alcoholic cardiomyopathy）的造模情况、优点和注意事项，C57或Balb/c小鼠喂酒精液体饲料容易建立ACM，而用转基因鼠例如ADH过表达小鼠或基因敲除小鼠例如金属硫蛋白基因敲除小鼠（metallothionein-knockout mice，MT-KO mice)喂酒精液体饲料可以进行特定机制的研究。

【关键词】心肌病模型；模型方法；大鼠；实验动物

酒精性心肌病（ACM，alcoholic cardiomyopathy）是一种扩张型心肌病（DCM，dilated cardiomyopathy），表现为心肌肥厚，心室肌收缩力下降，心输出量减少，心肌炎性细胞浸润，心脏纤维化。在人类患者可能与以下因素有关：（1）酒精（或酒精代谢产物乙醛：acetaldehyde）对心肌的毒性；（2）饮酒习惯导致饮食摄入热量和营养素（例如硫胺素B1,锌）不足而影响心脏功能；（3）酒精饮料中某些成分（例如钴摄入过多）的毒性。

但是，当建立“酒精性”（alcoholic）心肌病模型时，强调的就是(1)，消除（2）和（3），因为（2）和（3）不属于“酒精性”。即便有ACM表现，但包含（2）或（3）都是不可用的。

虽然小鼠的心脏小，但小鼠应用于建立酒精性心肌病的文献并不少。用小鼠进行ACM有几方面的优点：一是成本低，二是研究中可能用到的抗体来源丰富，几乎不受限制，三是不同基因背景的小鼠容易获得。

那么，小鼠建立酒精性心肌病的情况怎样呢，怎样复制和应当注意哪些方面？

一、小鼠酒精性心肌病造模的情况

早在1977年，当年有两篇研究论文使用C57B1小鼠进行了酒精液体饲料喂养试验，观察到喂15周后小鼠的心肌线粒体增多、肥大和融合成巨线粒体，甘油三酯聚积，闰盘断裂。25周时最明显的表现是间质水肿，肌纤维膜下脂肪聚积，间质纤维化。

不久前的一项研究中，给成年C57小鼠喂酒精液体饲料15周，表现出心肌肥厚，心肌收缩速度降低，细胞内钙浓度降低，细胞凋亡指标中Caspase-3表达上调，Bcl-2表达下调但JNK没有显著改变。

最近一项研究采用饮水中加酒精灌胃的方法为期90天，观察到Balb/c小鼠和C57小鼠的心功能指标发生显著改变，并且发生心肌纤维化。

二、转基因或基因敲除小鼠酒精性心肌病造模的情况

1. ADH（alcohol dehydrogenase）过表达的转基因小鼠。

该模型小鼠用于研究 acetaldehyde (ACA) 在酒精性心肌病中的作用。

ADH过表达的小鼠心肌酒精代谢增强，酒精摄入后心肌产生ACA（acetaldehyde）增多，因此，是一种研究酒精对心肌毒性和酒精性心肌病中ACA作用及其机制的动物模型。研究观察到ACA能够强力抑制心肌收缩能力。

2. 金属硫蛋白基因敲除小鼠（metallothionein-knockout mice，MT-KO mice)。

金属硫蛋白MT-I和MT-II亚单位的作用被消除后动物能够正常生长和繁殖，生理机能似乎没有明显改变。用酒精液体饲料喂养2个月心肌发生炎性细胞浸润、肥厚和纤维化，而野型小鼠（对照组）心肌没有异常表现，锌补充能够阻止MT-KO小鼠的纤维化，但对心肌肥厚没有作用。

3. 血管紧张素II的I型受体敲除小鼠（AT1 gene knockout mice，AT1-KO mice)

血管紧张素II对心肌功能发挥重要作用，对其受体（I型）敲除的小鼠进行酒精液体饲料喂养，观察ANG II在ACM中的作用和机理。

4. 胰岛素样生长因子I（IGF-1）过表达或IGF-1敲除小鼠

胰岛素样生长因子-1（insulin-like growth factor 1）进行敲除或过表达，观察这样的小鼠酒精液体饲料喂养后ACM的发生和发展及其机理。

三、心脏功能评估方法

小鼠心脏发生了哪些改变和造模是否达到预期要求，评估的内容方法包括：

1. 心动超声定量分析心脏解剖和心功能测定。

2. 离体心脏灌流法观察心脏功能。

3. 离体细胞培养分析。

4. 心脏重量、形态学观察和测量。

5. 生化和分子生物学方法分析。

关于上述方法的具体说明，请见“酒精性心肌病模型饲料和造模方法”。

这里需要说明的是，心动超声是比较理想的在体评估心脏功能的方法，可以观察心输出量，搏出量，心率，等等）和心脏解剖（心脏大小、心室大小、心室壁的厚度，室中膈的厚度等等）。然而，小鼠的心脏小，需要超声波仪器设备，特别是需要购置配套的特殊探头，因此，对实验室装备要求高。正因为如此，有的研究论文中没有采用，并不意味着不重要。

如果想了解大鼠如何建立ACM，请见：大鼠酒精性心肌病模型饲料和模型复制方法。如果想了解使用其他动物建立ACM的情况，请见：酒精性心肌病模型饲料和造模方法。

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小鼠酒精性心肌病(ACM)模型饲料和模型复制方法Alcoholic Cardiomyopathy Murine Model (ACM mouse Model) and Diet