瑞舒伐他订对大鼠脑出血后的Caspase-3,-7和c-IAP-1表达变化影响的研究

发布时间：2018-04-18 11:02

本文选题：脑出血 + Caspase-7　；参考：《右江民族医学院》2017年硕士论文

【摘要】：目的:通过观察在瑞舒伐他汀干预下大鼠实验性脑出血(Intracerebral hemorrhage,ICH)血肿周围脑组织中半胱氨酸蛋白酶3(cysteinylaspartate specific proteinases 3,Caspase-3)、半胱氨酸蛋白酶7(cysteinylaspartate specific proteinases 7,Caspase-7)和凋亡抑制蛋白1(cellular inhibitor of apoptosis proteins 1,c-IAP-1)的表达变化,探讨瑞舒伐他汀在脑出血发生后可能出现的神经保护作用及其机制,对临床用药起到一定指导作用。方法:(1)将60只SD大鼠按照随机分组的方法分为假手术组(A组,20只),脑出血组(B组,20只),瑞舒伐他汀干预组(C组,20只)。每组分成6h、24h、72h、5d四个时间点,每个时间点使用5只大鼠进行造模并实验。采用自体凝固血注入右侧尾状核法建立脑出血模型。(2)采用Garcia法对三组大鼠术后4个时间点分别进行神经缺损评分。(3)免疫组化法对三组大鼠血肿周围脑组织切片染色并在显微镜下观察分析。(4)RT-qPCR技术测定三组大鼠脑出血术后4个时间点血肿周围脑组织Caspase-3,-7及c-IAP-1表达状况。结果:(1)Garcia评分评估发现6小时时间点脑出血组和瑞舒伐他汀干预组相比差异无统计学意义,其余三个时间点瑞舒伐他汀干预组(C组)神经缺损情况较脑出血组(B组)减轻,P0.05,有统计学意义。两组均在72h神经缺损症状达到峰值,之后症状缓解。假手术组(A组)与其余两组在术后4个时间点相比发现假手术组(A组)神经缺损较轻,神经功能远高于其余两组,差异均有统计学意义。(2)免疫组化法染色后显微镜下可观察到假手术组(A组)4个时间点脑出血后c-IAP-1和Caspase-3,-7阳性细胞少量表达,脑出血组(B组)和瑞舒伐他汀干预组(C组)较假手术组(A组)每个时间点阳性细胞均出现显著的增高。除脑出血后6小时这一时间点外,其余各个时间点的瑞舒伐他汀干预组(C组)要比脑出血模型组(B组)c-IAP-1阳性细胞表达增多,Caspase-3,-7阳性细胞表达减少。瑞舒伐他汀能降低Caspase-3,-7及提高c-IAP-1水平(3)RT-qPCR法显示,假手术组组与脑出血模型组和瑞舒伐他汀组组术后各个时间点相比c-IAP-1及Caspase-3,-7表达水平低。6小时时间点脑出血组和瑞舒伐他汀干预组相比差异无统计学意义,其余时间点之间脑出血组和瑞舒伐他汀干预组相比,c-IAP-1表达降低,Caspase-3,-7表达增高,差异有统计学意义。结论:(1)瑞舒伐他汀可以降低脑出血后神经功能缺损症状。(2)瑞舒伐他汀能够抑制Caspase-3和Caspase-7活化,进而提高c-IAP-1的表达,c-IAP-1反作用于Caspase-3和Caspase-7,从而进一步抑制Caspase-3和Caspase-7表达。提示瑞舒伐他汀能够降低由细胞凋亡影响引起的继发性的脑损伤,对神经功能起到一定的保护作用。
[Abstract]:Objective: to observe the changes of the expression of Caspase-3, 7(cysteinylaspartate specific proteinases 7caspase-7 and 1(cellular inhibitor of apoptosis proteins 1c-IAP-1 in the perihematoma tissue of rat experimental intracerebral hemorrhage (ICH) induced by recuvastatin, and to observe the expression of 3(cysteinylaspartate specific proteinases 3 Caspase-3, 7(cysteinylaspartate specific proteinases 7caspase-7 and 1(cellular inhibitor of apoptosis proteins 1c-IAP-1.To explore the neuroprotective effect and mechanism of resuvastatin in patients with cerebral hemorrhage.Methods 60 Sprague-Dawley rats were randomly divided into sham-operated group (n = 20), intracerebral hemorrhage group (n = 20) and control group (n = 20).The rats in each group were divided into four time points (6 h, 24 h, 72 h and 5 d), and 5 rats were used at each time point to model and experiment.Establishment of intracerebral hemorrhage model by injecting autologous coagulation blood into right caudate nucleus. (2) Garcia method was used to evaluate the neurological defect of rats at 4 time points after operation.) Immunohistochemical method was used to stain the brain tissue sections around hematoma in three groups of rats.The expression of Caspase-3 + -7 and c-IAP-1 in brain tissue around intracerebral hemorrhage in three groups were detected by RT-PCR at four time points after intracerebral hemorrhage.Results there was no significant difference between the 6 hour cerebral hemorrhage group and the recuvastatin intervention group.At the other three time points, the nerve defect in the control group was significantly lower than that in the cerebral hemorrhage group (P 0.05).The symptoms of nerve defect reached the peak at 72 hours in both groups, and then relieved.Compared with the other two groups at 4 time points after operation, the sham operation group (group A) showed that the nerve defect in group A was lighter and the nerve function was much higher than that in the other two groups.The differences were statistically significant. (2) the expression of c-IAP-1 and Caspase-3 + -7 were observed under microscope in sham-operation group (group A) after intracerebral hemorrhage at four time points after immunohistochemical staining.The positive cells in group B (group B) and group C (group C) were significantly higher than those in group A (sham operation group) at each time point.Except for the time point of 6 hours after intracerebral hemorrhage, the expression of c-IAP-1 positive cells increased and the expression of Caspase-3 + -7 was decreased in the other time points of Risuvastatin intervention group (group C) than that in the model group of intracerebral hemorrhage (group B).Rosuvastatin could decrease Caspase-3 -7 and increase the level of c-IAP-1 by 3RT-qPCR.There was no significant difference in the expression of c-IAP-1 and Caspase-3 ~ (-7) between the sham operation group and the cerebral hemorrhage model group and the rosuvastatin group at different time points after operation compared with those of the cerebral hemorrhage group and the rosuvastatin intervention group.The expression of c-IAP-1 in ICH group and Risuvastatin group was significantly lower than that in control group at other time points.Conclusion Risuvastatin can reduce the neurological deficit symptoms after intracerebral hemorrhage. Resuvastatin can inhibit the activation of Caspase-3 and Caspase-7, and then increase the expression of c-IAP-1. C-IAP-1 counteracts Caspase-3 and Caspase-7, thus further inhibiting the expression of Caspase-3 and Caspase-7.These results suggest that rosuvastatin can reduce the secondary brain injury induced by apoptosis and play a protective role in neurologic function.
【学位授予单位】：右江民族医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.34

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