药物性亚冬眠对大鼠急性脑梗塞的影响

发布时间：2018-05-05 20:51

本文选题：药物性亚冬眠 + 冬眠合剂　；参考：《郑州大学》2013年硕士论文

【摘要】：目的通过观察药物性亚冬眠对大鼠急性脑梗塞脑组织中基质金属蛋白激酶-9(Matrix Metalloproteinase-9, MMP-9)和血小板活化因子(Platelet-Activating Factor,PAF)表达的变化,探讨药物性亚冬眠对大鼠急性脑梗塞的影响及可能的作用机制。方法成年雄性SD大鼠共63只,随机分成3组：对照组(只分离颈内、外动脉,不做栓塞处理)、模型组(通过改良Longa线栓法来制备大鼠大脑中动脉局灶性脑梗塞模型)及亚冬眠组(改良Longa线栓法造模成功后,给予冬眠合剂肌注),其中对照组及模型组在与亚冬眠组相同时间肌注等量生理盐水。三组分别于缺血后4h、12h、24h进行神经功能评分,各时间点取7只大鼠心脏灌洗后快速断头取脑进行HE染色,观察局部脑梗死的大体形态,并应用免疫组化Sp法检测PAF、MMP-9的表达。结果 1.神经功能缺损评分情况：模型组的大鼠神经功能缺损最为严重,其各时间点缺损评分情况与对照组相比较有显著性差异(P0.05);对照组大鼠无神经功能缺损,评分为0；而亚冬眠组大鼠神经功能缺损评分情况较模型组降低,其各时间点与模型组及对照组比较均有显著性差异(P0.05); 2.HE染色大体观察结果：模型组与亚冬眠组大鼠病灶脑组织的肿胀最为明显,亚冬眠组较模型组情况有所减轻,而对照组未见脑组织肿胀情况。进行HE染色之后在光镜下观察：对照组大鼠脑的海马区神经细胞能整齐排列,且有清晰的轮廓,密集,神经元胞浆丰富,淡染,胞核多位于细胞中央,核仁清晰；模型组海马区神经细胞轮廓模糊,排列不规则紊乱,数量减少,且间质水肿疏松,胞浆浓缩红染,分界不清,胞核皱缩或溶解甚至碎裂；亚冬眠组神经细胞与模型组相比结构尚清晰,间质水肿明显减轻,胞核与胞质尚能分清。 3.PAF、MMP-9免疫组化结果：各组均可见PAF、MMP-9阳性细胞表达,都在胞质中,呈现棕黄色染色,阳性细胞个数越多、染色越深,提示免疫反应越强,LOD值越大,则因子表达越多；在三组组内比较可见：各时间点PAF于脑缺血4h内已达较高水平,12h、24h值相对于4h未见明显差异(P0.05);MMP-9于脑缺血4h内表达已开始增高,于12h、24h持续升高。三组间比较可见：PAF、MMP-9于模型组及亚冬眠组各时间点与对照组比较均明显增高(P0.05);亚冬眠组各时间点与模型组相比,表达明显减少(P0.05)。结论药物性亚冬眠对大鼠急性脑梗塞损伤有脑保护作用,其作用机制可能通过减少钙离子过量内流、炎性介质的生成,进而减轻由此造成的代谢紊乱连锁反应的发生有关。
[Abstract]:Purpose To investigate the effect of drug subhibernation on the expression of matrix metalloproteinase-9 (MMP-9) and platelet activating factor Platelet-Activating factor-PAF (PAF) in rats with acute cerebral infarction, the effects of drug subhibernation on acute cerebral infarction and its possible mechanism were investigated. Method 63 adult male Sprague-Dawley rats were randomly divided into three groups: control group (internal and external carotid artery separation, Without embolization, the model group (model of focal cerebral infarction in the middle cerebral artery of rats by modified Longa thread occlusion method) and the subhibernating group (modified Longa thread embolization method were successfully established). The control group and model group were intramuscularly injected with normal saline at the same time as the subhibernation group. The neurological function scores were evaluated at 4 hours and 12 hours after ischemia in the three groups. The brain of 7 rats were quickly severed after heart lavage at each time point for HE staining. The gross morphology of local cerebral infarction was observed and the expression of PAFMP-9 was detected by immunohistochemical Sp method. Result 1. The neurological deficit score in the model group was the most serious, and there was significant difference between the model group and the control group (P 0.05). The score of nerve function defect in the subhibernation group was lower than that in the model group, and there was significant difference between the model group and the control group at each time point (P 0.05). The results of 2.HE staining showed that the swelling of brain tissue in the model group and the subhibernation group was the most obvious, and that in the subhibernating group was less than that in the model group, but there was no swelling in the brain tissue in the control group. After HE staining, the hippocampal neurons in the control group were neatly arranged, with clear outline, dense, rich cytoplasm and light staining. The nucleus was located in the center of the cell and the nucleolus was clear. In the model group, the contour of hippocampal neurons was fuzzy, the number of neurons was irregular, the number was decreased, the interstitial edema was loose, the cytoplasm was concentrated and red stain, the boundary was not clear, the nucleus was crumpled or dissolved or even broken. Compared with the model group, the nerve cells in the subhibernation group had clear structure, the interstitial edema was alleviated, and the nucleus and cytoplasm could be distinguished. 3. Immunohistochemical results of PAF- MMP 9: PAF- MMP 9 positive cells were expressed in cytoplasm. The more the number of positive cells was, the deeper the staining was, which indicated that the stronger the immune response was, the greater the expression of factors was. The comparison among the three groups showed that the expression of PAF at each time point reached a higher level within 4 h after cerebral ischemia than at 4 h after cerebral ischemia, and the expression of MMP 9 began to increase within 4 h after cerebral ischemia and continued to increase at 12 h after ischemia. Compared with the control group, the expression of MMP 9 in the model group and the subhibernation group was significantly higher than that in the control group, and the expression of MMP 9 in the subhibernation group was significantly lower than that in the model group. Conclusion Drug subhibernation has brain protective effect on acute cerebral infarction injury in rats. The mechanism may be related to reducing the formation of inflammatory mediators and reducing the chain reaction of metabolic disorder caused by calcium excess influx.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R743.33

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