阿托伐他汀對兔蛛網(wǎng)膜下腔出血后早期腦損傷的保護(hù)作用及其機制的實驗研究
發(fā)布時間：2018-08-09 11:10
【摘要】：目的:采用兔蛛網(wǎng)膜下腔出血后的動物模型,研究阿托伐他汀對兔蛛網(wǎng)膜下腔出血后早期海馬及腦干神經(jīng)元是否具有保護(hù)作用。方法:完全隨機將32只成年健康雄性新西蘭大白兔(2~2.5kg)分成SAH組(n=8)、對照組(n=8)、SAH+安慰劑組(n=8)及SAH+阿托伐他汀治療組(n=8)。應(yīng)用開顱枕大池一次注血法模擬蛛網(wǎng)膜下腔出血動物模型。分別于出血后4h、16h、28h、40h分別灌服劑量為5mg/kg/次的阿托伐他汀,48h后處死新西蘭大白兔。留取海馬、腦干做標(biāo)本。SAH建模成功后觀察新西蘭大白兔的行為功能變化,進(jìn)行神經(jīng)功能評分;分別采用TUNEL法檢測海馬、腦干神經(jīng)元的凋亡;同時采用免疫組化法測定Bcl-2、Bax因子表達(dá)的變化;結(jié)果:行為學(xué)觀察發(fā)現(xiàn)與對照組相比,SAH組新西蘭大白兔活動及飲食明顯減少,其平均神經(jīng)功能評分低,有顯著統(tǒng)計學(xué)意義(P0.01);阿托伐他汀治療組神經(jīng)功能評分優(yōu)于安慰劑組,差別有統(tǒng)計學(xué)意義(P0.05);凋亡細(xì)胞檢測顯示對照組中很少發(fā)現(xiàn)陽性細(xì)胞,SAH組陽性細(xì)胞明顯增多(P0.01),同時阿托伐他汀組陽性細(xì)胞數(shù)明顯少于安慰劑組,安慰劑組與SAH組陽性細(xì)胞數(shù)無差別(P0.05);凋亡相關(guān)因子免疫組化檢測發(fā)現(xiàn):SAH組與對照組比較,腦干及海馬區(qū)Bax、Bcl-2表達(dá)明顯增加(P0.05),阿托伐他汀組較安慰劑組Bax表達(dá)下降(P0.05),Bcl-2表達(dá)增加(P0.05)。安慰劑組較SAH組改善不顯著(P0.05)。結(jié)論:1.蛛網(wǎng)膜下腔出血后48可觀察到兔出現(xiàn)明顯神經(jīng)功能障礙并且腦干及海馬區(qū)神經(jīng)元出現(xiàn)明顯凋亡;2.阿托伐他汀可明顯抑制神經(jīng)元的凋亡,對早期腦干、海馬區(qū)神經(jīng)元具有保護(hù)作用。目的:利用兔蛛網(wǎng)膜下腔出血動物模型模型,研究SAH早期腦水腫與AQP4表達(dá)的關(guān)系,并探討阿托伐他汀對蛛網(wǎng)膜下腔出血早期腦水腫的影響及其機制。方法:32只成年健康雄性新西蘭大白兔(2~2.5kg)被隨機分成SAH組(n=8)、對照組(n=8)、SAH+安慰劑組(n=8)及SAH+阿托伐他汀治療組(n=8)。應(yīng)用開顱枕大池一次注血法模擬蛛網(wǎng)膜下腔出血動物模型。分別于出血后4h、16h、28h、40h分別灌服阿托伐他汀,劑量為5mg/kg/次,48h后處死新西蘭大白兔。留取整個大腦半球做標(biāo)本。采用干濕重法測定腦組織含水量;采用Western blot法測定AQP4的表達(dá)情況以及通過尼氏染色了解神經(jīng)元凋亡情況;結(jié)果:SAH后早期腦水腫明顯,SAH后48小時腦組織含水量達(dá)82.16%,同時測得AQP4的含量較高;與對照組比較腦組織含水量及AQP4的表達(dá)明顯增加(P0.01);阿托伐他汀治療組的含水量為80.13%,AQP4的含量較安慰劑組明顯下降(P0.01);而安慰劑組與對照組比較無明顯差別(P0.05);SAH組神經(jīng)元凋亡較對照組明顯增多而與安慰劑組比較無差異,而SAH+阿托伐他汀組神經(jīng)元凋亡較安慰劑組少,有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:1.蛛網(wǎng)膜下腔出血后早期即可產(chǎn)生腦水腫,且和AQP4的表達(dá)及神經(jīng)元凋亡呈正相關(guān);2.應(yīng)用阿托伐他汀治療蛛網(wǎng)膜下腔出血可明顯降低AQP4的表達(dá)且可改善血腦屏障減輕早期腦水腫。目的:利用蛛網(wǎng)膜下腔出血的動物模型,探討阿托伐他汀對蛛網(wǎng)膜下腔出血早期腦血管是否具有保護(hù)作用。方法:將24只成年健康雄性新西蘭大白兔(2~2.5kg)隨機分成SAH組(n=8)、對照組(n=8)及SAH+阿托伐他汀治療組(n=8)。應(yīng)用開顱枕大池一次注血法模擬蛛網(wǎng)膜下腔出血動物模型,對照組給予開顱去除顱骨但不注動脈血,SAH組開顱注入自體動脈血1ml/kg,阿托伐他汀組給予口服阿托伐他汀5mg/kg/次,分別于出血后4h、16h、28h、40h分別灌服阿托伐他汀,48h后處死新西蘭大白兔,留取基底動脈、顳葉皮層、腦干等組織做標(biāo)本,免疫組化法測定測定基底動脈中v WF及TM的表達(dá)情況,RT-PCR測定血管內(nèi)皮細(xì)胞中v WF及TM的m RNA的表達(dá),HE染色測定基底動脈血管內(nèi)徑周長與管壁厚度比值即T值(D/A)。結(jié)果:RT-PCR結(jié)果顯示對照組的v WF及TM的m RNA的表達(dá)水平較弱,SAH組較對照組比較其v WF及TM的m RNA的表達(dá)明顯增加(P0.01);阿托伐他汀治療組v WF及TM的m RNA的表達(dá)明顯低于SAH組(P0.01)。免疫組化結(jié)果顯示對照組中v WF及TM的表達(dá)陽性率較低;與對照組相比,SAH組v WF及TM的陽性率明顯增加;阿托伐他汀治療組v WF及TM的表達(dá)陽性率低于SAH組。HE染色結(jié)果顯示SAH組較對照組比較,基底動脈內(nèi)徑周長與管壁厚度比(T值)明顯降低(P0.01);阿托伐他汀組T值明顯高于SAH組(P0.01)。結(jié)論:1.SAH早期即可引起腦血管痙攣以及血管內(nèi)皮細(xì)胞自我調(diào)節(jié)功能障礙;2.阿托伐他汀可以保護(hù)血管內(nèi)皮細(xì)胞,減輕血管痙攣;3.阿托伐他汀保護(hù)血管內(nèi)皮細(xì)胞以及促進(jìn)血管內(nèi)皮細(xì)胞增生的機制可能與抑制v WF及TM的表達(dá)有關(guān)。
[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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R743.35
本文編號：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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R743.35
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