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Medicine
心肌缺血再灌注损伤的改进啮齿动物模型
心肌缺血再灌注损伤的改进啮齿动物模型
JoVE Journal
Medicine
This content is Free Access.
JoVE Journal Medicine
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury

心肌缺血再灌注损伤的改进啮齿动物模型

Full Text
6,543 Views
07:23 min
March 7, 2022

DOI: 10.3791/63510-v

Hua-Qin Tong*1,2,3, Man-Lu Fan*3, Tong Sun3, Hao-Wen Zhang4, Jie Han5, Meng-Xi Wang3, Jian-Dong Chen5, Wei-Xin Sun6, Xiao-Hu Chen1,2, Mian-Hua Wu7

1Department of Cardiology,Affiliated Hospital of Nanjing University of Chinese Medicine, 2Department of Cardiology,Jiangsu Province Hospital of Chinese Medicine, 3First College of Clinical Medicine, Biological Technology Center for Innovation in Chinese Medicine,Nanjing University of Chinese Medicine, 4School of Health Preservation and Rehabilitation,Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, 5Department of Cardiology,Jiangsu Provincial Hospital of Chinese Medicine, 6Department of Cardiology,Yancheng TCM Hospital Affiliated to Nanjing University of Chinese Medicine, 7Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor,Nanjing University of Chinese Medicine

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

Summary

通过使用自制的牵开器,聚氯乙烯管和独特的打结方法,改善了大鼠心脏的心肌缺血再灌注模型。心电图、三苯基氯化四唑鎓和组织学染色以及生存百分比分析结果表明,改进的模型组比已有的模型组具有更高的成功率和存活率。

Transcript

这种心肌缺血再灌注模型通过自制的牵开器,聚氯乙烯管和独特的打结方法得到改进。这种技术与更高的存活率和更大的冲击大小有关。该方法将提高大鼠心肌缺血再灌注损伤的生存率,为未来心肌缺血再灌注损伤的临床治疗带来启示。

麻醉大鼠后,将两个回形针的中间部分拉直以形成S形,并拉下每个S的宽部分以形成一个小的牵开器。将一根直径为两毫米的聚氯乙烯管切成七毫米长的片。将10厘米长的4-0缝合线插入PVC管并绑住其末端。

然后用眼科剪刀在PVC管的中间切一个凹槽,在结扎期间放置缝合线。通过将大鼠以仰卧姿势放置在手术板上,用胶带固定大鼠的四肢。用脱毛霜剃除颈部和左侧前胸部区域,并用75%酒精和碘伏磨砂膏清洁皮肤。

然后用眼科剪刀沿正中颈线纵向切割颈部皮肤。使用眼科镊子分离颈部肌肉,并在每侧放置一个牵开器以进一步缩回它们。暴露气管后,确定第四和第五气管环之间的空间。

在第四和第五气管环之间用针尖的钝边,在第四和第五气管环之间做一个平行于环状软骨的三毫米切口。通过切口将吸盘插入气管中,并对大鼠进行机械通气,以每分钟80次呼吸和每公斤8毫升的潮气量维持正常呼吸。接下来,从xiphoid到第二个左肋间间隙的中间做一个四到五厘米长的切口,同时以45度角握住脚手架。

使用眼科镊子轻轻缓慢地分离胸大肌和锯齿前肌进入肋间间隙。使用眼科剪刀在左肋骨、第三肋骨和第四肋骨之间横向切开 1.5 厘米的切口。为了防止受伤,将浸泡在胸腔肺部上方生理盐水溶液中的棉球。

使用眼用镊子解剖心包。然后用镊子抬起左心耳,识别主动脉根部存在的冠状动脉口。在左肺和耳廓之间的部分,使用6-0手术缝合线将LAD和预先准备好的短管连接在一起。

使用滑结将它们绑起来,并将滑结放在PVC管的凹槽中。然后使用第二个滑动结拧紧结扎管和LAD 45分钟。在缺血期间,在心电图上记录左心室前部的颜色变化和ST段抬高。

使用动脉夹夹住胸部肌肉和皮肤,用湿润的盐水纱布覆盖伤口,然后松开滑结,并在45分钟后取出预先准备好的短管。接受现有或改善心肌缺血和再灌注损伤程序的大鼠有心肌梗塞,而假组的大鼠没有。此外,现有和实验性心肌缺血和再灌注损伤模型组在心肌梗死大小方面存在显著差异。

实验模型组的心肌梗死大小大于现有模型组。苏木精,曙红和Masson染色表明,实验组和现有模型组的心肌细胞在被大量嗜中性粒细胞浸润时都经历了严重的损伤和核细胞增多。现有和实验性心肌缺血和再灌注损伤模型组的大鼠比假组显示心电图STT段升高。

实验模型与假组或现有模型与假组存在显著差异。此外,实验模型组中的TT段比现有模型组的TT段更高。心肌缺血和再灌注损伤模型组的存活率差异有统计学意义。

再灌注期间的死亡率为40%,而在实验模型组中,没有一只大鼠在手术过程中死亡,这表明当前改进的模型具有更高的存活率。最重要的是,我们应该把管子放进结里。同时,将滑结放入管子的凹槽中。

这种方法还可以深入了解心肌缺血再置换损伤和心肌感染领域。

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