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Medicine
一种改良的小鼠心肌梗死诱导简易方法
一种改良的小鼠心肌梗死诱导简易方法
JoVE Journal
Medicine
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JoVE Journal Medicine
A Modified Simple Method for Induction of Myocardial Infarction in Mice

一种改良的小鼠心肌梗死诱导简易方法

Full Text
4,093 Views
04:29 min
December 3, 2021

DOI: 10.3791/63042-v

Chenglin Jiang*1,2, Jianqing Chen*1,2, Yichao Zhao*2, Dasheng Gao1, Hongju Wang1, Jun Pu1,2

1Department of Cardiology,The First Affiliated Hospital of Bengbu Medical College, 2Department of Cardiology, Ren Ji Hospital, School of Medicine,Shanghai Jiao Tong University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Summary

在充分麻醉下,小鼠心脏通过肋间腔外化,并使用大多数实验室中现成的材料结扎左前降支 (LAD) 成功诱导心肌梗死。

Transcript

本报告提出了一种诱导小鼠心肌梗死的独特方法。目前的方法简单、省时,并且使用大多数实验室中容易找到的手术管和设备。心肌梗死模型的建立为研究心肌梗死后重塑的既往机制和新疗法的变异提供了前提。

将脱毛膏涂抹在小鼠胸部,等待一分钟。然后用湿纱布轻轻擦去脱毛膏和头发。将鼠标固定在仰卧位的手术平台上。

按照文中所述对手术区域进行消毒。然后在消毒区域周围放置无菌窗帘。沿着剑突和腋窝的连接线切开0.5厘米的皮肤,用镊子钝性地分开胸大肌和胸小肌。

使用Micro-Mosquito止血器打开第四肋间空间。用左手食指将心脏推向第四肋间隙,将心脏外化。用左手固定心脏,并用 6/0 缝合线将左前降支结扎在距其原点 3 毫米处。

迅速将心脏放回胸腔。轻轻按压胸腔,将空气排出胸腔。对于术后疼痛管理,每 4 至 6 小时皮下注射一次丁丙诺啡,持续术后长达 72 小时。

将鼠标放在加热垫上以从麻醉中恢复。将安乐死的小鼠固定在手术平台上的仰卧位。在上腹部做一个大约三到四厘米的腹侧切口。

从胸腔两侧切掉肋骨,然后取下横膈膜。通过心室内注射用 10 毫升冷的 4 摄氏度 1X PBS 灌注心脏。然后通过切断主动脉根部来收集心脏,并立即将其储存在零下 80 摄氏度。

使用剃须刀片将冷冻的心脏在冰上切成一毫米厚的部分。将准备好的心脏切片在溶解在 37 摄氏度的 1X PBS 中的 1% TTC 溶液中孵育 10 至 15 分钟。然后使用数码相机拍摄切片。

冠状动脉结扎术后,TTC染色分析显示心肌梗死诱导成功,心肌梗死后瘢痕大小随时间变化。生存分析结果显示,雄性和雌性C57BL/6J小鼠在心肌梗死后7天内出现明显死亡。心室破裂是心肌梗死后死亡的常见原因。

心肌梗死后超声心动图评估显示成功诱导收缩功能障碍和心室重塑。当试图将心脏外化时,不要先按压胸腔,跟随我们的抵抗,表明需要调整方向以对抗加权的赞美。在此程序之后,可以进行病毒载体或细胞的心肌内注射,以测试基因疗法或细胞疗法在心脏病方面的疗效。

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改良简单方法 心肌梗死诱导 心肌梗死模型 心肌梗死后重塑 新型疗法 异丙肾上腺素治疗 冷冻损伤 冠状动脉结扎 缺血反应 慢性重塑 技术挑战 现成材料 小鼠胸皮 肋间隙 胸大肌 胸小肌 左前降支 (LAD) 6-0 缝合线 2 3 5-三苯基氯化四氮唑 (TTC) 疤痕大小变化 生存分析结果 心脏破裂 超声心动图评估

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