一种基于血管内皮损伤的局灶性脑梗死血瘀证大鼠模型的建立

【作者】 陈懿；

【导师】 邓冰湘； 葛金文；

【作者基本信息】 湖南中医学院 ， 中西结合基础， 2002， 硕士

【摘要】 目的：本实验旨在通过损伤脑血管内皮细胞来建立一种新型的大鼠局灶性脑梗死血瘀证模型。 方法：选择健康SD大鼠58只，随机分为模型组（model group）大鼠28只、假手术组（simulation group）大鼠21只和正常对照组（control group）大鼠9只。模型组向左颈内动脉注入月桂酸钠150ug，假手术组注入等体积生理盐水，正常对照组不作任何处理。 模型组、假手术组在术后6、12和24小时三个时相各处死大鼠7只，正常对照组在实验开始后处死大鼠6只，用于行为体征观察、血浆tPA和PAI活性检测、TXB₂和6-Keto-PGF_1α含量测定以及脑组织梗死灶大小评估，脑组织病理形态学观察。模型组在术后6小时另处死大鼠7只，正常对照组实验开始后另处死大鼠3只，用于血液流变学指标检测。 结果：①缺血6小时后可出现不同程度的神经功能缺失体征，但在12小时或24小时观察，发现其症状逐渐减弱，功能逐渐恢复。 ②通过TTC染色可见，大鼠正常脑组织呈红色，在缺血6小时后、大脑皮质外侧部、皮质下的尾状核、内囊、下丘脑的外侧部表现出明显的缺血梗死区，梗死区呈苍白色。缺血12、24小时后，梗死区范围未见扩大。 ③肉眼观察，梗死侧比对侧苍白，脑组织较软。光镜下观察：缺血6小时后，皮质及海马区肿胀明显，组织间隙增宽，并有部分神经元出现坏死，细胞核固缩，破裂。缺血12、24小时后，缺血区与6小时相比变化不大。 ④颈内动脉注射后各时间点，假手术组和正常对照组各项指标无差异；但模型组在注入月桂酸钠后 6d’时血浆 tPA活性明显低于假手术组和正常对照组（P叩．05），而＊I活性则明显高于其它各组(P硼．0U，tPA与 PAI活性f匕值在术后 6，J、时和 1卜J、时较低，与正常对照组和假手术组比较有差异（P叩．0 5）；术后2 4，J、时 f匕值升高，与 6、12，1、时间差异显著（P叩。01）。模型组术后6，］、时uB；含量最高，与其他各组比较有差异(P叩．0v；而 6《；大小PGF；。在各组间没有明显的差异。 ；⑤模型组注入月桂酸纳6小时后，血液粘度、全血还原粘度及血浆粘度升高，与正常组相比较差异非常显著（P叩．OS）；但红细胞压积在两组间比较无差异O叩．“L 结论：一定量的月桂酸钠可诱导大民局灶性脑梗死血瘀证发生，表现为脑组织缺血坏死，血管内皮一血小板功能紊乱，纤溶功能明显改变，血液流变性异常。该模型的建立可望为研究血管内皮细胞在脑梗死发生发展中的作用及中医药防治脑梗死提供一种重要的实验工具。更多还原

【Abstract】 Objective: A new rat model of cerebral infarction with blood stasis syndrome was developed to elucidate the contribution of vascular endothelial cell during focal cerebral cerebral infarction.Methods: fifty-eight SD rats were randomly divided into model group with 28 rats in which sodium laurate was injected into the left internal carotid artery of the anesthetized rats, simulation group with 21 rats in which sodium laurate was substituted with 0.9% sodium chloride solution, and control group with 9 rats(no treament). 6, 12, 24 hours after operation, 7 rats were respectively executed for index observation in model and simulation group, as well as 6 rats in control group, such as triplienyltetrazoliurn chloride(TTC) dyeing, neurological dificit, plasma tissue-type plasminogen activator(tPA) activity, plasminogen activator inhibitor(PAI) activity, thromboxaneB2 (TXB2) content, 6-keto.-PGF1 n content, histopathological analyses. In addition, 7 rats of model group at 6 hour after operation and 3 rats of control group were used for hemorheology inspection.Results: (1)In a postural reflex test, rats were tested for degree of abnormal posture. Neurological score was the highest at 6 hour after operation, descending significantly in the later period. (2)6hours after operation, infarct brain regions were not stained, showing white determined by TTC in the lateral cortex, subcortical caudatum, internal capsule and lateral hypothalarnus, and the infarct size did not expand with experiment course. On the other hand, no-ischemic regions were stained red. (3) Infarct region wasobserved pale and soft compared with contralater hemisphere by naked eye. The results were gained under optical microscope, including cortex neuron and hippocampus swelling, cells distance extending, cells neurosis, et al. (4)The activity of tPA at 6 hour after operation was significantly lower than that in simulation and control groups(P<0.05), while PAI activity significantly higher than others’(P<0.05). 6 hour after operation plasma TXB2 content was the highest compared with other groups(P<0.05), but no distinction concerning plasma 6-Keto-PGF1 content was detected in all groups. (5) There was obvious difference between control and model groups on blood viscosity, reduction viscosity and plasmaviscosity.Conclusion: Focal cerebral infarction blood stasis syndrome in rats could be induced by some sodium laurate, showing ischemic cerebrum necrosis., function disorder of vascular endothelium-platelet, fibrinolysis and hemorheology abnormity. This model could be an important method for researching the contribution of vascular endothelial cell during cerebral infarct development, prevention and cure by tradition Chinese medicine.更多还原