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Neuroscience
用于研究创伤性脑损伤出血和炎症的刺伤小鼠模型
用于研究创伤性脑损伤出血和炎症的刺伤小鼠模型
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Stab-Wound Mouse Model for Studying Hemorrhage and Inflammation in Traumatic Brain Injury

用于研究创伤性脑损伤出血和炎症的刺伤小鼠模型

Full Text
729 Views
04:03 min
February 21, 2025

DOI: 10.3791/67797-v

Kei Hashimoto1, Mari Nakashima1, Yasunori Miyamoto1, Hiroko Ikeshima-Kataoka2

1Institute for Human Life Science,Ochanomizu University, 2Kitasato University School of Frontier Engineering

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

Summary

简化的创伤性脑损伤 (TBI) 模型促进了治疗方法的发展。该协议概述了使用针头创建刺伤小鼠皮层,从而能够分析出血和炎症。刺伤性 TBI 小鼠模型的优势在于无需专用设备即可执行。

Transcript

我们有兴趣开发一种止血和抑制 TVI 炎症的方法,因此我们建立了一种方案,只需一根针即可轻松制作 TVI 质量模型。我们的刺伤 TVI 模型是有益的,因为它可以润滑出血和灰细胞活化。与其他技术相比,我们的协议有三个主要优势。

不需要特定的设备或工具。它可以轻松快速地完成,而且伤口非常小,不会影响鼠标的行为,因此任何人都可以以高重现性执行此作。我们的实验室将专注于神经元、灰细胞和神经胶质细胞以及创伤性脑损伤修复过程中寄生虫的通讯。

沟通在维修中起着至关重要的作用。然而,到目前为止,人们还没有完全理解它。我们的协议对于分析通信非常有用。

首先,将麻醉的小鼠放在纸巾的腹侧。通过脚趾捏试验确认鼠标已深度麻醉。通过交替轮次 Betadine 和 70% 乙醇擦洗来消毒手术部位,每次 3 次。

现在,用钝钳抓住枕骨皮肤,做一个 1 到 1.5 毫米宽的切口,露出枕骨,而不会损坏颅骨或任何器官。然后轻轻缓慢地打开切口,通过颅骨观察大脑皮层和小脑之间的边界。要进行小鼠开颅手术,请用针在枕骨的右半球开一个小孔。

轻轻旋转针头,使针刺伤口成为插入点,注意不要损伤脑实质。对于刺伤,从插入点插入合适的针头,并沿喙尾轴刺入大脑皮层。最后,使用带有半圆形针头的尼龙 3-0 缝合线缝合皮肤切口。

手术后的伤口通过显微镜检查确认。IGG 外渗在受伤后 1 天达到峰值,随后进行性恢复,并在 3 、 5 和 7 天染色强度降低。Evan 的蓝色染料在受伤后立即渗漏到脑实质中,其浓度在 1 小时时达到峰值。

染料浓度在 1 d 和 3 d 后逐渐降低。IBA1 阳性小胶质细胞和 GFAP 阳性星形胶质细胞的积累高峰分别发生在损伤后 5 天和 3 天,它们的存在减少了 7 天。炎性细胞因子,包括肿瘤坏死因子 α、白细胞介素 6 和白细胞介素 1 β 在受伤后 1 天显示 mRNA 表达峰值,而转化生长因子 β 1 在受伤后 3 天达到峰值。

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创伤性脑损伤 TBI 小鼠模型 刺伤模型 出血 炎症 神经元丢失 原发性损伤 继发性损伤 脑挫伤 颅骨骨折 细胞因子产生 神经胶质细胞活化 治疗剂

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