阿托伐他汀对兔蛛网膜下腔出血后早期脑损伤的保护作用及其机制的实验研究
发布时间：2018-08-09 11:10
【摘要】：目的:采用兔蛛网膜下腔出血后的动物模型,研究阿托伐他汀对兔蛛网膜下腔出血后早期海马及脑干神经元是否具有保护作用。方法:完全随机将32只成年健康雄性新西兰大白兔(2~2.5kg)分成SAH组(n=8)、对照组(n=8)、SAH+安慰剂组(n=8)及SAH+阿托伐他汀治疗组(n=8)。应用开颅枕大池一次注血法模拟蛛网膜下腔出血动物模型。分别于出血后4h、16h、28h、40h分别灌服剂量为5mg/kg/次的阿托伐他汀,48h后处死新西兰大白兔。留取海马、脑干做标本。SAH建模成功后观察新西兰大白兔的行为功能变化,进行神经功能评分;分别采用TUNEL法检测海马、脑干神经元的凋亡;同时采用免疫组化法测定Bcl-2、Bax因子表达的变化;结果:行为学观察发现与对照组相比,SAH组新西兰大白兔活动及饮食明显减少,其平均神经功能评分低,有显著统计学意义(P0.01);阿托伐他汀治疗组神经功能评分优于安慰剂组,差别有统计学意义(P0.05);凋亡细胞检测显示对照组中很少发现阳性细胞,SAH组阳性细胞明显增多(P0.01),同时阿托伐他汀组阳性细胞数明显少于安慰剂组,安慰剂组与SAH组阳性细胞数无差别(P0.05);凋亡相关因子免疫组化检测发现:SAH组与对照组比较,脑干及海马区Bax、Bcl-2表达明显增加(P0.05),阿托伐他汀组较安慰剂组Bax表达下降(P0.05),Bcl-2表达增加(P0.05)。安慰剂组较SAH组改善不显著(P0.05)。结论:1.蛛网膜下腔出血后48可观察到兔出现明显神经功能障碍并且脑干及海马区神经元出现明显凋亡;2.阿托伐他汀可明显抑制神经元的凋亡,对早期脑干、海马区神经元具有保护作用。目的:利用兔蛛网膜下腔出血动物模型模型,研究SAH早期脑水肿与AQP4表达的关系,并探讨阿托伐他汀对蛛网膜下腔出血早期脑水肿的影响及其机制。方法:32只成年健康雄性新西兰大白兔(2~2.5kg)被随机分成SAH组(n=8)、对照组(n=8)、SAH+安慰剂组(n=8)及SAH+阿托伐他汀治疗组(n=8)。应用开颅枕大池一次注血法模拟蛛网膜下腔出血动物模型。分别于出血后4h、16h、28h、40h分别灌服阿托伐他汀,剂量为5mg/kg/次,48h后处死新西兰大白兔。留取整个大脑半球做标本。采用干湿重法测定脑组织含水量;采用Western blot法测定AQP4的表达情况以及通过尼氏染色了解神经元凋亡情况;结果:SAH后早期脑水肿明显,SAH后48小时脑组织含水量达82.16%,同时测得AQP4的含量较高;与对照组比较脑组织含水量及AQP4的表达明显增加(P0.01);阿托伐他汀治疗组的含水量为80.13%,AQP4的含量较安慰剂组明显下降(P0.01);而安慰剂组与对照组比较无明显差别(P0.05);SAH组神经元凋亡较对照组明显增多而与安慰剂组比较无差异,而SAH+阿托伐他汀组神经元凋亡较安慰剂组少,有统计学意义(P0.05)。结论:1.蛛网膜下腔出血后早期即可产生脑水肿,且和AQP4的表达及神经元凋亡呈正相关;2.应用阿托伐他汀治疗蛛网膜下腔出血可明显降低AQP4的表达且可改善血脑屏障减轻早期脑水肿。目的:利用蛛网膜下腔出血的动物模型,探讨阿托伐他汀对蛛网膜下腔出血早期脑血管是否具有保护作用。方法:将24只成年健康雄性新西兰大白兔(2~2.5kg)随机分成SAH组(n=8)、对照组(n=8)及SAH+阿托伐他汀治疗组(n=8)。应用开颅枕大池一次注血法模拟蛛网膜下腔出血动物模型,对照组给予开颅去除颅骨但不注动脉血,SAH组开颅注入自体动脉血1ml/kg,阿托伐他汀组给予口服阿托伐他汀5mg/kg/次,分别于出血后4h、16h、28h、40h分别灌服阿托伐他汀,48h后处死新西兰大白兔,留取基底动脉、颞叶皮层、脑干等组织做标本,免疫组化法测定测定基底动脉中v WF及TM的表达情况,RT-PCR测定血管内皮细胞中v WF及TM的m RNA的表达,HE染色测定基底动脉血管内径周长与管壁厚度比值即T值(D/A)。结果:RT-PCR结果显示对照组的v WF及TM的m RNA的表达水平较弱,SAH组较对照组比较其v WF及TM的m RNA的表达明显增加(P0.01);阿托伐他汀治疗组v WF及TM的m RNA的表达明显低于SAH组(P0.01)。免疫组化结果显示对照组中v WF及TM的表达阳性率较低;与对照组相比,SAH组v WF及TM的阳性率明显增加;阿托伐他汀治疗组v WF及TM的表达阳性率低于SAH组。HE染色结果显示SAH组较对照组比较,基底动脉内径周长与管壁厚度比(T值)明显降低(P0.01);阿托伐他汀组T值明显高于SAH组(P0.01)。结论:1.SAH早期即可引起脑血管痉挛以及血管内皮细胞自我调节功能障碍;2.阿托伐他汀可以保护血管内皮细胞,减轻血管痉挛;3.阿托伐他汀保护血管内皮细胞以及促进血管内皮细胞增生的机制可能与抑制v WF及TM的表达有关。
[Abstract]:Objective: To study the protective effect of atorvastatin on the early hippocampal and brainstem neurons in rabbits after subarachnoid hemorrhage after subarachnoid hemorrhage in rabbits. Methods: 32 adult healthy New Zealand white rabbits (2~2.5kg) were randomly divided into SAH group (n=8), control group (n=8), SAH+ placebo group (n=8) and SAH+ opioid. The treatment group of atorvastatin (n=8) was used to simulate the animal model of subarachnoid hemorrhage with one time injection of cranioccipital big pool and blood injection. After bleeding, 4h, 16h, 28h, 40H were administered to atorvastatin at a dose of 5mg/kg/ times respectively. After 48h, New Zealand white rabbits were executed. The hippocampus was retained and the brain stem was successfully established to observe the behavior work of New Zealand white rabbits. TUNEL method was used to detect the apoptosis of the hippocampus and the brain stem neurons, and the changes in the expression of Bcl-2 and Bax were measured by immunohistochemistry. The results showed that compared with the control group, the activity and diet of New Zealand white rabbits in the group SAH were significantly reduced, and the average neurological function score was low and there was a significant difference. Study significance (P0.01); the neurologic score of the Atorvastatin group was better than that in the placebo group (P0.05); apoptotic cell detection showed that there were few positive cells in the control group, and the positive cells in the group SAH were significantly increased (P0.01), and the number of positive cells in the Atorvastatin group was significantly less than that in the placebo group, and the placebo group and the SAH group were significantly less. The number of positive cells was not different (P0.05); the immunohistochemical detection of apoptosis related factors found that the expression of Bax and Bcl-2 in the brain stem and hippocampus increased significantly in SAH group (P0.05). The expression of Bax in the Atorvastatin group was lower than that in the placebo group (P0.05) and Bcl-2 expression increased (P0.05). The placebo group was not significantly improved in the SAH group (P0.05). Conclusion: 1. subarachnoid membrane (P0.05). After intracerebral hemorrhage, 48 can be observed in rabbits with obvious nerve dysfunction and obvious apoptosis in the brain stem and hippocampus neurons. 2. ataratartin can obviously inhibit neuronal apoptosis and have protective effects on the early brain stem and hippocampus neurons. Objective: To study the early brain edema and A in SAH with the animal model model of subarachnoid hemorrhage in rabbits. The relationship between QP4 expression and the effect of atorvastatin on early cerebral edema of subarachnoid hemorrhage and its mechanism. Methods: 32 healthy adult male New Zealand rabbits (2~2.5kg) were randomly divided into SAH group (n=8), control group (n=8), SAH+ placebo group (n=8) and SAH+ atrovastatin group (n=8). A single injection of cranioccipital big pool was used. The animal model of subarachnoid hemorrhage was simulated. After hemorrhage, 4h, 16h, 28h, and 40H were respectively administered to atorvastatin, the dose was 5mg/kg/, and the New Zealand white rabbit was killed after 48h. The whole brain hemisphere was left to be taken as a specimen. The water content of the brain tissue was measured by the dry wet weight method. The expression of AQP4 was measured by Western blot method and the Nissl staining was used. The results were as follows: the brain edema was obvious in the early stage after SAH, the water content of brain tissue was 82.16% in the 48 hours after SAH, and the content of AQP4 was higher than that in the control group (P0.01), and the water content of the Atorvastatin group was 80.13%, and the content of AQP4 was significantly lower than that of the placebo group (P0. 01) but there was no significant difference between the placebo group and the control group (P0.05); the neuron apoptosis in the SAH group was significantly higher than that in the control group, and there was no difference between the placebo group and the placebo group, while the neuron apoptosis in the SAH+ atrovastatin group was less than that in the placebo group, and was statistically significant (P0.05). Conclusion: cerebral edema can be produced early after the subarachnoid hemorrhage, and AQP4 There is a positive correlation between expression and neuronal apoptosis; 2. the treatment of subarachnoid hemorrhage with atorvastatin can significantly reduce the expression of AQP4 and improve the blood brain barrier to reduce early cerebral edema. Objective: To explore the protective effect of atorvastatin on the early cerebral vessels of subarachnoid hemorrhage by using the animal model of subarachnoid hemorrhage. Method: 24 adult healthy male New Zealand white rabbits (2~2.5kg) were randomly divided into group SAH (n=8), control group (n=8) and SAH+ atorvastatin treatment group (n=8). The animal model of subarachnoid hemorrhage was simulated with one time blood injection of cranioccipital big pool, and the control group was given craniotomy but no arterial blood, and SAH group craniotomy was injected into autologous blood 1ml/kg. The Atorvastatin group was given oral atorvastatin 5mg/kg/ times. After hemorrhage 4h, 16h, 28h, and 40H were administered to atorvastatin, respectively, and after 48h, the New Zealand white rabbits were sacrificed, and the basilar artery, the temporal cortex and the brain stem were left to be specimens. The expression of V WF and TM in the base artery was measured by immunohistochemical method, and RT-PCR was used to determine the vascular endothelium. The expression of M RNA of V WF and TM in the cell, the ratio of the circumference of the basilar artery to the thickness of the tube wall was T value (D/A) by HE staining. Results: RT-PCR results showed that the expression level of V WF and TM was weaker than that of the control group. The expression of NA was significantly lower than that of the SAH group (P0.01). The positive rate of V WF and TM in the control group was lower than that in the control group, and the positive rate of V WF and TM in the SAH group was significantly higher than that in the control group, and the positive rate of V WF and expression in the Atorvastatin group was lower than that of the control group, and compared with the control group, the circumference of the basilar artery was compared with the control group. The thickness ratio of tube wall (T) was significantly lower (P0.01); the T value of atorvastatin group was significantly higher than that in group SAH (P0.01). Conclusion: early 1.SAH could cause cerebral vasospasm and vascular endothelial cell self-regulation dysfunction; 2. atorvastatin could protect vascular endothelial cells, reduce vascular spasm, and 3. atorvastatin protected vascular endothelial cells. And the mechanism of promoting the proliferation of vascular endothelial cells may be related to the inhibition of the expression of V WF and TM.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R743.35
本文编号：2173888
[Abstract]:Objective: To study the protective effect of atorvastatin on the early hippocampal and brainstem neurons in rabbits after subarachnoid hemorrhage after subarachnoid hemorrhage in rabbits. Methods: 32 adult healthy New Zealand white rabbits (2~2.5kg) were randomly divided into SAH group (n=8), control group (n=8), SAH+ placebo group (n=8) and SAH+ opioid. The treatment group of atorvastatin (n=8) was used to simulate the animal model of subarachnoid hemorrhage with one time injection of cranioccipital big pool and blood injection. After bleeding, 4h, 16h, 28h, 40H were administered to atorvastatin at a dose of 5mg/kg/ times respectively. After 48h, New Zealand white rabbits were executed. The hippocampus was retained and the brain stem was successfully established to observe the behavior work of New Zealand white rabbits. TUNEL method was used to detect the apoptosis of the hippocampus and the brain stem neurons, and the changes in the expression of Bcl-2 and Bax were measured by immunohistochemistry. The results showed that compared with the control group, the activity and diet of New Zealand white rabbits in the group SAH were significantly reduced, and the average neurological function score was low and there was a significant difference. Study significance (P0.01); the neurologic score of the Atorvastatin group was better than that in the placebo group (P0.05); apoptotic cell detection showed that there were few positive cells in the control group, and the positive cells in the group SAH were significantly increased (P0.01), and the number of positive cells in the Atorvastatin group was significantly less than that in the placebo group, and the placebo group and the SAH group were significantly less. The number of positive cells was not different (P0.05); the immunohistochemical detection of apoptosis related factors found that the expression of Bax and Bcl-2 in the brain stem and hippocampus increased significantly in SAH group (P0.05). The expression of Bax in the Atorvastatin group was lower than that in the placebo group (P0.05) and Bcl-2 expression increased (P0.05). The placebo group was not significantly improved in the SAH group (P0.05). Conclusion: 1. subarachnoid membrane (P0.05). After intracerebral hemorrhage, 48 can be observed in rabbits with obvious nerve dysfunction and obvious apoptosis in the brain stem and hippocampus neurons. 2. ataratartin can obviously inhibit neuronal apoptosis and have protective effects on the early brain stem and hippocampus neurons. Objective: To study the early brain edema and A in SAH with the animal model model of subarachnoid hemorrhage in rabbits. The relationship between QP4 expression and the effect of atorvastatin on early cerebral edema of subarachnoid hemorrhage and its mechanism. Methods: 32 healthy adult male New Zealand rabbits (2~2.5kg) were randomly divided into SAH group (n=8), control group (n=8), SAH+ placebo group (n=8) and SAH+ atrovastatin group (n=8). A single injection of cranioccipital big pool was used. The animal model of subarachnoid hemorrhage was simulated. After hemorrhage, 4h, 16h, 28h, and 40H were respectively administered to atorvastatin, the dose was 5mg/kg/, and the New Zealand white rabbit was killed after 48h. The whole brain hemisphere was left to be taken as a specimen. The water content of the brain tissue was measured by the dry wet weight method. The expression of AQP4 was measured by Western blot method and the Nissl staining was used. The results were as follows: the brain edema was obvious in the early stage after SAH, the water content of brain tissue was 82.16% in the 48 hours after SAH, and the content of AQP4 was higher than that in the control group (P0.01), and the water content of the Atorvastatin group was 80.13%, and the content of AQP4 was significantly lower than that of the placebo group (P0. 01) but there was no significant difference between the placebo group and the control group (P0.05); the neuron apoptosis in the SAH group was significantly higher than that in the control group, and there was no difference between the placebo group and the placebo group, while the neuron apoptosis in the SAH+ atrovastatin group was less than that in the placebo group, and was statistically significant (P0.05). Conclusion: cerebral edema can be produced early after the subarachnoid hemorrhage, and AQP4 There is a positive correlation between expression and neuronal apoptosis; 2. the treatment of subarachnoid hemorrhage with atorvastatin can significantly reduce the expression of AQP4 and improve the blood brain barrier to reduce early cerebral edema. Objective: To explore the protective effect of atorvastatin on the early cerebral vessels of subarachnoid hemorrhage by using the animal model of subarachnoid hemorrhage. Method: 24 adult healthy male New Zealand white rabbits (2~2.5kg) were randomly divided into group SAH (n=8), control group (n=8) and SAH+ atorvastatin treatment group (n=8). The animal model of subarachnoid hemorrhage was simulated with one time blood injection of cranioccipital big pool, and the control group was given craniotomy but no arterial blood, and SAH group craniotomy was injected into autologous blood 1ml/kg. The Atorvastatin group was given oral atorvastatin 5mg/kg/ times. After hemorrhage 4h, 16h, 28h, and 40H were administered to atorvastatin, respectively, and after 48h, the New Zealand white rabbits were sacrificed, and the basilar artery, the temporal cortex and the brain stem were left to be specimens. The expression of V WF and TM in the base artery was measured by immunohistochemical method, and RT-PCR was used to determine the vascular endothelium. The expression of M RNA of V WF and TM in the cell, the ratio of the circumference of the basilar artery to the thickness of the tube wall was T value (D/A) by HE staining. Results: RT-PCR results showed that the expression level of V WF and TM was weaker than that of the control group. The expression of NA was significantly lower than that of the SAH group (P0.01). The positive rate of V WF and TM in the control group was lower than that in the control group, and the positive rate of V WF and TM in the SAH group was significantly higher than that in the control group, and the positive rate of V WF and expression in the Atorvastatin group was lower than that of the control group, and compared with the control group, the circumference of the basilar artery was compared with the control group. The thickness ratio of tube wall (T) was significantly lower (P0.01); the T value of atorvastatin group was significantly higher than that in group SAH (P0.01). Conclusion: early 1.SAH could cause cerebral vasospasm and vascular endothelial cell self-regulation dysfunction; 2. atorvastatin could protect vascular endothelial cells, reduce vascular spasm, and 3. atorvastatin protected vascular endothelial cells. And the mechanism of promoting the proliferation of vascular endothelial cells may be related to the inhibition of the expression of V WF and TM.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R743.35
【参考文献】
相关期刊论文 前4条
1 涂建飞;刘一之;纪建松;;血管内皮细胞凋亡在兔脑血管痉挛模型的初步研究[J];介入放射学杂志;2013年07期
2 刘殿玮;姜勇;韩向阳;;蛛网膜下腔出血后下丘脑caspase-3、bax和bcl-2的表达及意义[J];山东大学学报(医学版);2010年02期
3 邢红霞;李建国;杨晋生;赵新利;;阿托伐他汀治疗急性脑出血脑水肿作用机制的探讨[J];山东医药;2007年01期
4 马朝晖;李贵福;罗望池;黄燕;李铁林;;改良“二次枕大池注血法”构建Wistar大鼠蛛网膜下腔出血模型[J];中国神经精神疾病杂志;2012年03期
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