电凝法制作C57/BL6J小鼠局灶性脑缺血模型的评价  

Validation of a C57/BL6J mouse model of focal cerebral ischemia established by electrocoagulation of the middle cerebral artery

作　　者：康赟赟 唐东宁 张健[1,2] 夏青[1,2] KANG Yunyun;TANG Dongning;ZHANG Jian;XIA Qing(Research Center of Experimental Acupuncture,Tianjin University of Chinese Medicine,Tianjin 301617,China;School of Medical Technology,Tianjin University of Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学实验针灸学研究中心,天津301617 [2]天津中医药大学医学技术学院,天津301617

出　　处：《南方医科大学学报》2024年第1期100-107,共8页Journal of Southern Medical University

基　　金：国家自然科学基金青年科学基金(82105021)。

摘　　要：目的通过改良电凝法制作小鼠局灶性大脑中动脉阻塞(MCAO)模型的方法、筛选适用于模型小鼠的评价指标,为开展脑缺血相关的长期研究提供模型参考。方法选用C57/BL6J雄性小鼠46只,随机分为模型组(n=34)和对照组(n=12)。采用凝血器永久性凝断小鼠右侧大脑中动脉制备大脑中动脉闭塞模型。通过激光散斑血流成像仪监测造模前、造模后20min、造模后1d的脑血流灌注量;通过Longa评分、mNSS评分、平衡木行走实验、圆筒实验、转角实验评价小鼠造模后1、7、14d的神经功能;采用2,3,5-氯化三苯基四氮唑(TTC)染色测量缺血灶梗死范围,以评估模型是否成功及各指标之间的相关性。Western blotting检测缺血侧皮质中质纤维酸性蛋白(GFAP)、脑源性神经营养因子(BDNF)、双肾上腺皮质激素(DCX)蛋白表达。结果激光散斑血流成像仪监测显示,大脑中动脉电凝灼烧后缺血侧的脑血流量明显降低;与假手术组相比,模型组造模后1、7、14d的Longa评分和mNSS评分明显升高(P<0.05);平衡木行走实验结果显示,与假手术组相比,模型组造模后1、7、14d评分均降低(P<0.05);圆筒实验结果显示,与术前相比,造模后1、7、14d模型组的前肢使用不对称性均增加(P<0.05);转角实验结果显示,与术前相比,造模后1、7、14d模型组的侧向指数均有提高(P<0.05)。TTC染色显示,假手术组大脑组织无损伤,模型组的梗死灶在手术侧大脑的感觉运动皮层区和尾壳核。Westernblotting结果显示,与假手术组相比,模型组缺血侧脑皮层中GFAP的蛋白表达水平呈高表达,模型组1d(P<0.05)、模型组7d(P<0.01)、模型组14d(P<0.01)的GFAP蛋白表达水平升高;DCX的蛋白表达水平在造模后1、7、14d升高(P<0.05);BDNF的蛋白表达水平升高,模型组1d的BDNF蛋白表达水平在缺血后显著升高(P<0.05)。结论将电凝位置由MCA与大脑下静脉交界的远端位置改为MCA与嗅束交界内侧2mm段�Objective To modify the method for establishing mouse models of middle cerebral artery occlusion(MCAO)-induced focal cerebral ischemia using electrocoagulation.Methods Forty-six C57/BL6J male mice were divided into MCAO model group(n=34)and sham-operated group(n=12).In the model group,MCAO was induced by permanent coagulation of the right middle cerebral artery(MCA)using a coagulator,and cerebral blood flow perfusion was monitored before and at 20 min and 1 day after modeling.Neurological deficits of the mice at 1,7,and 14 days after modeling were evaluated using Longa score,mNSS score,beam walking test,cylinder test and corner test.TTC staining was used to measure the cerebral infarct size,and Western blotting was performed to detect the expressions of BDNF,GFAP and DCX proteins in the ischemic cortex.Results The mice in the model group showed significantly reduced cerebral blood flow in the MCA on the ischemic side and obvious neurological deficits with increased forelimb use asymmetry on days 1,7 and 14 after modeling(P<0.05).In the cerebral cortex on the ischemic side of the model mice,the expressions of GFAP and DCX increased significantly at 1,7,and 14 days(P<0.05)and the expression of BDNF increased at 1 day after modeling ischemia(P<0.05).Conclusion We successfully prepared mouse models of MCAO using a modified method by changing the electrocoagulation location from the distal location of the junction between the MCA and the inferior cerebral vein to a 2 mm segment medial to the junction between the MCA and the olfactory bundle.

关 键 词：电凝法 大脑中动脉 脑缺血小鼠 评价指标 

分 类 号：R743.31[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]