Improved suture occlusion method to prepare focal cerebral ischemia reperfusion models in Sprague-Dawley rats

doi:10.3969/j.issn.2095-4344.2014.02.011

Abstract

Abstract:

BACKGROUND: During the progress of focal cerebral ischemia-reperfusion in rat with transient suture occlusion, bleeding combating and the line being plugged into the cerebral middle artery smoothly are important for successful modeling and research focus.
OBJECTIVE: To improve Longa suture occlusion method for focal cerebral ischemia reperfusion model in Sprague-Dawley rats in order to establish a simple surgical procedure with low rate of intraoperative bleeding and high rate of successful models.
METHODS: The two ends of the external carotid artery were clipped with electric knife, in addition to ligation of surgery lines intraoperatively; with the help of pincett, a fishing line was plugged into the carotid artery along the anteriomedialis artery wall (the direction of the internal carotid artery from the pterygopalatine arterial bifurcation). Postoperative behavior scores in rats, infarct volume calculation and histopathological evaluation under optical microscope were done.
RESULTS AND CONCLUSION: Transverse sections of the external carotid artery were changed from a “O” shape to a “-” shape after being clipped with electric knife. Subsequently, line nodes were not easy to fall off any more, effectively preventing intraoperative bleeding. With the power of pincett, the rate of plugging the fishing line into artery was significantly increased. Rat’s neurological score and infarct volumes were significantly increased, and brain tissue pathological injury worsened. The modified suture occlusion method for focal cerebral ischemia reperfusion models effectively prevented line nodes off, successfully increased the rate of suture occlusion and models in Sprague-Dawley rats were developed successfully.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: models, animal, reperfusion injury, carotid artery, external, rats, Sprague-Dawley

CLC Number:

R318

Sun Nian-xia, Gao Wei-juan, Yi Zhong-liang. Improved suture occlusion method to prepare focal cerebral ischemia reperfusion models in Sprague-Dawley rats[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(2): 225-230.

Figures/Tables 1

2.1 实验动物数量分析假手术组大鼠全部纳入结果分析，按造模成功标准改良模型组26只大鼠纳入结果分析，Longa模型组17只大鼠纳入结果分析，改良模型组成模率(87%)与Longa模型组成模率(60%)比较差异有显著性意义(P < 0.05)(图2A)。 2.2 TTC染色和脑梗死体积改良模型组脑梗死体积占大脑总体积的(13.66±6.01)%，Longa模型组脑梗死体积占大脑总体积的(12.89±5.74)%，均与假手术组(0%)比较差异有显著性意义(P < 0.001)，两模型组间差异无显著性意义(P > 0.05)(图2B)。假手术组大鼠冠状切面左右脑半球染色均匀、对称，灰质呈红色，白质呈浅红或瓷白色，无呈苍白色的梗死区(图2C1)。模型组大鼠脑组织缺血侧可见皮质和(或)皮质下呈苍白色的梗死区域，包括基底核的外侧、顶叶和颞叶皮质、海马、纹状体等区域，与对侧半球呈红色的正常脑组织对比鲜明，白色区与红色区域之间界限不明显(图2C2，C3)。 2.3 组织病理学观察结果光镜下假手术组海马区各层细胞排列规则、整齐，核和核仁清晰可见，胞质内尼氏体为紫蓝色颗粒或块状结构，神经突起自然排列(图3A)。模型组术侧海马区各层细胞排列紊乱，部分神经细胞消失，残存神经细胞胞膜皱缩内陷，神经纤维异常弯曲，胞质内尼氏体消失、模糊，核固缩均质化，核仁易位(图3B，C)。假手术组各叶皮质细胞层次清晰，核均匀，核仁清晰、圆形、居中，胞质内尼氏体丰富、深染，胞体有多个突起(图3D)。模型组缺血侧顶叶、颞叶皮质梗死区和正常区之间无明显界限，梗死区神经细胞出现缺血性损害，部分神经细胞消失，残存神经细胞结构紊乱，纤维弯曲，胞膜外周出现空泡，胞膜皱缩凹陷，胞浆内尼氏体模糊、淡染，核固缩、边集，可见胶质细胞吞噬坏死神经元的现象，间质水肿，见图(图3E，F)。"

References

[1] 陈茉弦,敖丽娟,李琦,等.脑卒中动物模型的建立与比较[J].中国康复医学杂志,2011,26(8):779-782.

[2] Ma J, Zhao L, Nowak TS Jr.Selective, reversible occlusion of the middle cerebral artery in rats by an intraluminal approach. Optimized filament design and methodology.J Neurosci Methods. 2006;156(1-2):76-83.

[3] 胡凌,高培毅.线栓法制备大鼠局灶性脑缺血模型影响因素的探讨[J].中国医学影像技术,2004,20(10):1624-1626.

[4] 王耀明,董萼塘,肖学宏.MRI评价改进的大鼠可逆性局灶性脑缺血模型[J].卒中与神经疾病,1999,6(3):171-173.

[5] 戴炯,文立,熊文浩,等.改良线栓法制备大鼠局灶性脑缺血/再灌注模型[J].上海交通大学学报:医学版,2007,27(10): 1218-1222.

[6] 龚彪,李长清,黄剑.SD大鼠线栓法局灶性脑缺血/再灌注模型的改进[J].重庆医学,2006,35(4):313-315.

[7] Liu C, Weaver J, Liu KJ.Rapid conditioning with oxygen oscillation: neuroprotection by intermittent normobaric hyperoxia after transient focal cerebral ischemia in rats.Stroke. 2012;43(1):220-226.

[8] Heydenreich N, Nolte MW, Göb E,et al. C1-inhibitor protects from brain ischemia-reperfusion injury by combined antiinflammatory and antithrombotic mechanisms.Stroke. 2012;43(9):2457-2467.

[9] Lu T, Luo Y, Sun H,et al. Electroacupuncture improves behavioral recovery and increases SCF/c-kit expression in a rat model of focal cerebral ischemia/reperfusion.Neurol Sci. 2013;34(4):487-495.

[10] Longa EZ, Weinstein PR, Carlson S,et al. Reversible middle cerebral artery occlusion without craniectomy in rats.Stroke. 1989;20(1):84-91.

[11] 黄茸茸,李卫平,李维祖.黄芪提取物对大鼠局灶性脑缺血再灌注血脑屏障损伤的影响[J].中国现代应用药学,2010,27(12): 1064- 1068.

[12] 涂悦,姚旻,陈炳伟,等.不同浓度TTC染色对高血压性脑卒中病灶面积的评估[J].武警医学院学报,2008,17(1):30-32,49.

[13] Koizumi J,Yoshida Y,Nakazawa T,et al. Experimental studies of ischemic brain edema. 1. A new experimental model of cerebral embolism infarcts in the ischemic area. Japan J Stroke. 1986;(8):1-8.

[14] Longa EZ, Weinstein PR, Carlson S,et al. Reversible middle cerebral artery occlusion without craniectomy in rats.Stroke. 1989;20(1):84-91.

[15] 彭拥军,陈理,郭景春,等.改良大鼠脑缺血再灌注模型制作方法[J].辽宁中医药大学学报,2010,12(11):48-50.

[16] 王棕可,胡跃强.大鼠脑缺血再灌注模型改良的探讨[J].中国中医急症,2011,20(8):1286-1287.

[17] 牛建国,文玉军,张莲香,等.大鼠大脑中动脉阻塞再灌注模型改良的实验研究[J].宁夏医学院学报,2008,30(1):7-10,13.

[18] 朱继,万东,唐文渊.改良线栓法制备大鼠局灶脑缺血模型[J].第四军医大学学报,2008,29(8):685-687.

[19] 沈斌, Xu Haoliang.线栓法制备SD大鼠局灶性脑缺血/再灌注模型的改良研究[J].首都医科大学学报,2007,28(6):782-784.

[20] 吴浩,吉训明,赵喜庆,等.改良大鼠MCAO模型的建立[J].中华实验外科杂志,2006,23(12):1553-1554.

[21] 李浩,贾建平.大鼠大脑中动脉线栓法的改良与评价[J].中风与神经疾病杂志,2006,23(6):666-668.

[22] 吴松笛, 耿晓英,丁桃英,等.改良线栓法制作大鼠局灶性脑缺血再灌注模型的实验研究[J].陕西医学杂志,2006,35(10): 1316- 1318.

[23] 徐立新,袁芳.改良的大鼠脑局部缺血模型[J].中国康复理论与实践,2005,11(1):23-24.

[24] 吉训明,凌锋,赵喜庆,等.改良可逆性局灶性脑缺血模型建立[J].介入放射学杂志,2005,14(2):178-181.

[25] 胡凌,高培毅.改良线栓法制备大鼠脑局部低灌注模型的研究[J].中国医学影像技术,2004,20(8):1182-1185.

[26] 孙宇,严国锋,姜虹,等.改良线栓法大鼠大脑中动脉阻塞模型的建立[J].中国比较医学杂志,2008,18(8):8-10.

[27] Kuge Y, Minematsu K, Yamaguchi T, et al. Nylon monofilament for intraluminal middle cerebral artery occlusion in rats.Stroke. 1995;26(9):1655-1657.

[28] 徐旭东,刘文彦,孙秀丽,等.大鼠局灶性脑缺血动物模型的实验研究[J].济宁医学院学报,2000,23(1):9-11.

[29] Abrahám H, Somogyvári-Vigh A, Maderdrut JL,et al. Filament size influences temperature changes and brain damage following middle cerebral artery occlusion in rats.Exp Brain Res. 2002;142(1):131-138.

[30] 李忠仁,崔龙.TC指数在改良LONGA线栓法局灶性脑缺血/再灌注动物模型制备中的应用[J].中国中西医结合杂志,2006,26(S1): 18-20.

[31] 汪飞,储照虎,赵守才.线栓法大鼠大脑中动脉闭塞模型的制备[J].基层医学论坛,2011,15(1):14-17.

[32] 赵凯,张国华,贾宇臣,等.线栓法制备大鼠局灶性脑缺血/再灌注模型的改良及探讨[J].中国卒中杂志,2012,7(12):916-921.

[33] 刘亢丁.实验性局灶性脑缺血再灌注动物模型的改进及评价[J].中风与神经疾病杂志,1997,14(2):87.

[34] 潘江,章薇,葛卫军,等.改良永久性线栓法制作大鼠大脑中动脉梗阻模型的探讨[J].湖南中医药大学学报,2011,31(3):24-26,44.

[35] 吴远华,朱广旗,胡蓉,等.线栓法大鼠脑缺血再灌注模型改良与评价[J].中国实用神经疾病杂志,2010,13(18):4-6.

[36] 李向荣,韩江全,何敏.改良法制作SD大鼠局灶性脑缺血/再灌注模型探讨[J].遵义医学院学报,2008,31(5):486-487.

[37] 杨德兵,张利宏,马存根.大鼠大脑中动脉缺血再灌注模型的改良[J].中国民族民间医药,2009,18(16):27-28.

[38] 陈春富,李劲松.线栓法大鼠脑缺血模型的研究[J].中风与神经疾病杂志,1996,13(1):18-19.

[39] Robinson RG.Differential behavioral and biochemical effects of right and left hemispheric cerebral infarction in the rat. Science. 1979;205(4407):707-710.