rhG-CSF对大鼠脑缺血超时间窗溶栓后MMP-9表达及出血转化的影响

发布时间：2018-05-27 15:08

本文选题：脑梗死 + 模型　；参考：《山西医科大学》2016年硕士论文

【摘要】：第一部分目的:选取合适线栓直径,建立符合后续实验研究需要的实验模型。方法:选取体重250±20g健康雄性SD大鼠20只,随机分为A、B两组(n=10),实验操作采用改良线栓法造成大鼠右侧大脑中动脉闭塞(MCAO)模型,A组线栓直径0.26mm,B组0.30mm,于6小时后拔出线栓再灌注,24小时后行Longa神经功能缺损评分及TTC法测脑梗死体积并计算百分比。结果描述主要以sx±表示,采用t检验及卡方检验,P0.05认为差异具有统计学意义。结果:1.两组大鼠缺血再灌注后均表现出神经功能缺损:走路时一侧行走不利,身体倾斜向一侧或原地划圈打转,精神低靡,活动较前明显减少,自主饮食摄水减少。Longa评分A组(2.4±0.69)分,B组(3.1±0.73)分,存在明显统计学差异(P0.05)。2.A组10只大鼠缺血再灌注后全部存活,B组3只大鼠表现为意识丧失,1只死亡,1-3分表明造模成功,经卡方检验,两组之间造模成功率A组高于B组。3.脑切片TTC染色后可见正常脑组织部分呈红色,而梗死区域呈现白色。A组(35.89±3.33)%,B组(39.69±4.44)%,结果显示:两组大鼠脑梗死体积百分比A组小于B组。结论:体重250±20g大鼠线栓法缺血6小时再灌注造模时,选用直径0.26mm的鱼线所造模型成功率更高,可以造成稳定的局灶性脑梗死,为后续实验的顺利进行提供一定的模型基础。第二部分目的:探讨人重组粒细胞集落刺激因子(rh G-CSF)对大鼠脑缺血超时间窗(缺血6小时)溶栓治疗后基质金属蛋白酶-9(MMP-9)表达及并发症出血转化发生的影响,为扩大溶栓时间窗提供理论依据。方法:选取健康雄性SD大鼠36只,按第一部分制备MCAO模型,随机分为3组,即空白对照组、重组组织型纤溶酶原激活剂(rt-PA)组及联合治疗组(n=12),缺血后6h三组给予拔除线栓处理,同时给药,rt-PA组给予rt-PA 10 mg/kg,联合治疗组给予rt-PA(10 mg/kg)+rh G-CSF(300μg/kg),空白对照组仅给等容积生理盐水。再灌注24h后行神经功能缺损评分,免疫组化法测定MMP-9表达,光谱光度法测量脑出血量。统计描述主要以sx±表示,定量资料采用t检验和方差分析,定性资料采用卡方检验。P0.05认为差异具有统计学意义。结果:1.三组大鼠造模后均表现出神经功能缺损,分别给予各组处理后神经功能缺损评分空白对照组(2.25±0.622)分,rt-PA组(2.08±0.515)分,联合治疗组(1.33±0.651)分,经LSD-t检验,结果显示:联合治疗组与其他两组的Longa评分值均有明显的统计学差异(P0.05)。2.MMP-9表达:非缺血侧半球罕见MMP-9免疫阳性细胞表达,缺血半球侧则表达较多,呈棕黄色淡染。各组缺血侧与正常对侧情况比较结果表明:三组大鼠病变侧MMP-9表达高于正常对侧。缺血侧阳性细胞计数空白对照组24.33±2.733,rt-PA组30.00±2.280,联合治疗组20.83±1.722,LSD-t检验结果表明:rt-PA组MMP-9表达较空白组明显增加,联合治疗组MMP-9表达较rt-PA组下降。3.三组大鼠脑出血量比较结果显示:rt-PA组脑出血量较空白对照组明显增加,而联合治疗组较rt-PA组出血量明显减少(P0.05)。结论:rh G-CSF降低大鼠超时间窗(6h)溶栓后出血转化的发生,可能与rh G-CSF降低MMP-9表达有关。
[Abstract]:The first part was to select the appropriate diameter of the thread and establish the experimental model in accordance with the needs of the follow-up experiment. Methods: 20 rats with weight 250 20g healthy male SD rats were selected and randomly divided into A and B two groups (n=10). The experimental operation by modified thread embolus was used to cause the right middle cerebral artery occlusion (MCAO) model in rats, the diameter of the A group, the B group 0.30mm, and the A group. 6 hours after pulling out the thread thrombus and reperfusion, after 24 hours, the Longa nerve function defect score and the TTC method were used to measure the cerebral infarction volume and calculate the percentage. The results were described mainly by SX +, using t test and chi square test, P0.05 thought the difference was statistically significant. Results: 1. two groups of rats showed nerve function defect after blood reperfusion: walking On one side, the body was disadvantageous, the body sloped to one side or the original area, the spirit was low, the activity was reduced obviously, the.Longa score in A group (2.4 + 0.69) and B group (3.1 + 0.73) were reduced, and there was significant statistical difference (P0.05) in group.2.A, 10 rats were all survived after ischemia and reperfusion, and 3 rats in group B showed loss of consciousness, 1 Only death, 1-3 points showed the success of the model, after the chi square test, the success rate of the two groups was higher than the B group.3. brain slice TTC staining, the normal brain tissue was red, and the infarct area showed white.A group (35.89 + 3.33)%, B group (39.69 + 4.44)%. The results showed that the percentage of cerebral infarction volume A group in two groups was less than that of group B. Conclusion: weight 250 in group two. Conclusion: weight 250. Conclusion: body weight 250. Conclusion: weight 250 in group two. Conclusion: body weight 250 is less than that of group B. Conclusion: weight 250 of group of cerebral infarction in two groups is less than that of group B. Conclusion: weight 250 of group.3. is less than that of group B. Conclusion: weight 250 in group of two rats is smaller than that of group B. Conclusion: weight 250 of group.3. is less than that of group B. Conclusion: weight 250 In the 6 hour reperfusion model of 6 hour ischemia in the rats of + 0.26mm, the success rate of the model with the diameter of 0.26mm is higher, which can cause a stable focal cerebral infarction and provide a certain model basis for the smooth progress of the follow-up experiment. The purpose of this study is to explore the human recombinant granulocyte colony-stimulating factor (RH G-CSF) for the cerebral ischemia in rats The effect of time window (ischemia 6 hours) on the expression of matrix metalloproteinase -9 (MMP-9) and complication of hemorrhage transformation after thrombolytic therapy, in order to provide a theoretical basis for expanding the thrombolytic time window. Methods: 36 healthy male SD rats were selected and the MCAO model was prepared in the first part, which were randomly divided into 3 groups, that is, the blank control group and the recombinant tissue type plasminogen activator. The rt-PA group and the combined treatment group (n=12), after the ischemia, the 6h three groups were treated with the extraction line thrombus, at the same time, the drug was given, the rt-PA group was given rt-PA 10 mg/kg, the combined treatment group was given rt-PA (10 mg/kg) +rh G-CSF (300 mu g/kg), the blank control group was only given equal volume physiological saline. The amount of cerebral hemorrhage was measured by spectral photometric method. The statistical description was mainly expressed in SX. The quantitative data were analyzed by t test and variance analysis. The qualitative data were statistically significant by chi square test.P0.05. Results: 1. the three groups of rats showed nerve function defect after the model was made, and the scores of nerve function defect were given to each group respectively. The group (2.25 + 0.622), rt-PA group (2.08 + 0.515), combined treatment group (1.33 + 0.651) score and LSD-t test showed that the Longa score of the combined treatment group and the other two groups had significant statistical difference (P0.05).2.MMP-9 expression: the expression of MMP-9 immunoreactive cells in the non ischemic hemisphere was rare, and the ischemic hemisphere side was more expressed in brown. The comparison between the ischemic side and the normal contralateral situation showed that the expression of MMP-9 in the three groups was higher than that in the normal contralateral side. The positive cells in the ischemic side were 24.33 + 2.733 in the blank control group, 30 + 2.280 in the group rt-PA and 20.83 in the combined treatment group. The results of LSD-t test showed that the expression of MMP-9 in the group rt-PA was significantly higher than that in the blank group. The combination of the MMP-9 expression in the group rt-PA was significantly higher than that in the blank group. Compared with group rt-PA, the comparison of MMP-9 expression in the group of.3. three rats showed that the amount of cerebral hemorrhage in group rt-PA was significantly higher than that in the blank control group, while the amount of hemorrhage in the combined treatment group was significantly lower than that in the rt-PA group (P0.05). Conclusion: RH G-CSF reduced the occurrence of hemorrhage after thrombolytic thrombolysis in the super time window (6h) of rats, and may reduce the MMP-9 table with RH G-CSF. It's up to it.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R743.3

【相似文献】