醒脑静对大鼠急性颅脑损伤后脑水肿的治疗作用及机制的研究
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摘要
研究目的
通过建立sD大鼠自由落体脑创伤模型,观察腹腔注射XNJ对大鼠急性颅脑创伤后脑水肿、BBB的损害及AQP4的影响,探讨其脑保护作用及作用机制。
研究方法
1.实验动物
清洁级、健康、成熟sD雄性大鼠108只,体重300～350g。随机分为假手术组、脑外伤组和XNJ治疗组。后两组再分为伤后1、3、5、7d等4个亚组,每亚组12只。
2.实验方法
采用参照Feeney'S自由落体模型设计并适当改进的打击装置。用20 g重的击锤从30 cm高处自由坠落冲击撞杆,冲击力为600 g·cm,造成脑挫裂伤,制作大鼠脑创伤模型。XNJ治疗组在大鼠脑外伤模型做好后10 min内腹腔内注射XNJ 10mL·kg~(-1),每天一次。脑外伤组则腹腔内给予等量0.9%氯化钠溶液。假手术组仅行开颅术,不造成脑损伤。
3.标本采集和检测
3.1脑组织含水量检测将大鼠伤后第1、3、5、7d分别断头处死后,立即取出伤侧大鼠脑组织,110℃恒温箱烘烤24 h至恒重。用干湿质量法计算脑组织含水量。计算公式为:脑组织含水量(%)=(湿质量-干质量)/湿质量×100%。假手术组术后24 h做同样处理。
3.2 BBB定量测定采用EB比色法。各组于处死前1 h通过股静脉注射2%EB(3mL·kg~(-1))。取脑组织前,快速开胸暴露心脏,迅速穿刺左心室后,剪右心房一小口,用37℃生理盐水500 mL灌注,至右心房流出清亮液体。取伤灶周边脑组织约0.2 g,加人5 mL甲酰胺,45℃水浴24 h,1500 r·min~(-1)离心10 min,取上清液用HP8453型分光光度计(λ=635 nm)测其光密度值。用倍比稀释法制作标准曲线,再根据标准曲线求得脑组织EB含量,结果以μg·g~(-1)(脑组织)表示。
3.3 AQP4免疫组化染色各组大鼠在预定时间点在伤灶区周围取脑组织,4%甲醛固定24-48 h,石蜡包埋,然后切片行HE染色和免疫组化检查。以细胞膜呈棕黄色或褐色为AQP4阳性染色。应用NIKON彩色病理图文分析系统进行图像分析。
3.4脑组织AQP4 mRNA的实时荧光RT-PCR测定分别取各组不同时间点的损伤周围脑组织进行RT-PCR分析,按Trizol试剂说明提取组织总RNA,逆转录为cDNA,以逆转录所得eDNA为模板进行PCR扩增。AQP4引物:上游5′GGT CCT CAT CTC CCTCTG CTT 3′,下游5′AAC CGT GGT GAC TCC CAA TCC3′,扩增片段长270 bp;β-肌动蛋白引物:上游5′TGT GAT GGT GGG TAT GGG 3′,下游5′TAG AAG CATTTG CGG TGC 3′,扩增片段长度为241 bp。反应体系50 uL,PCR条件:95℃预变性3 min;再95℃变性15 s,58℃退火15 s,72℃延伸30 s,共40个循环。求得各基因的Ct值。采用2~(-ΔΔCt)方法,即以管家基因β-肌动蛋白的Ct值标准化目的基因Ct值,得到各时点目的基因初始模板相对量,再计算各时点目的基因初始模板相对量与假手术组目的基因初始模板相对量的比值,分析各时点目的基因AQP4mRNA表达相对量的变化。
4.统计学处理
采用SPSS11.5统计软件包进行处理,资料以均数±标准差((?)±s)表示,采用单因素方差分析方法(One-Way ANOVA);P＜0.05为差异有统计学意义。
研究结果
1.各组大鼠脑含水量比较
脑外伤组脑组织含水量伤后1d达高峰,并持续至第3d,第5d开始下降,第7天仍明显高于假手术组(P＜0.05)。XNJ组脑含水量也在第1d达高峰,后逐渐下降,但均明显低于脑外伤组(P＜0.05);并于第7d降至接近正常,与假手术组比较无显著性差异(P＞0.05)。
2.各组大鼠脑组织EB含量比较
脑外伤组EB含量伤后1d达高峰,后逐渐下降,各时点均明显高于假手术组(P＜0.05)。XNJ组EB含量伤后1d达高峰,也逐渐下降,各时点均明显低于脑外伤组(P＜0.05),但均不同程度高于假手术组。
3.脑含水量与EB的相关分析
脑含水量与EB的变化规律相同,相关分析得,r=0.898,P＜0.01,表明二者具有正性相关。
4.AQP4免疫组化染色和AQP4 mRNA
光学显微镜下观察,以细胞膜呈棕黄色或褐色为AQP4阳性染色。脑外伤组伤后1d AQP4免疫染色变深,它的mRNA水平上调,并持续至第3d,明显高于假手术组(P＜0.05);在第5d开始下降,与假手术组比较无显著性差异(P＞0.05)。XNJ治疗后1、3 d的AQP4和AQP4 mRNA较脑外伤组明显上调(P＜0.05)。
结论
XNJ可以减轻创伤性脑水肿,可能与其减轻BBB破坏,上调AQP4表达有关。
Objective
In order to clarify the protective effects and the mechanism of XNJ,TBI rat models were established,brain water content,BBB permeability,AQP4 and AQP4 mRNA were tested. Methods
1.Subjects
108 male Sprague-Dawley rats approximately 300-350g,were used as subjects and randomly assigned to three groups:sham-operation,TBI and XNJ group.The later two groups were divided into 4 subgroups at 1,3,5 and 7d.
2.Models
TBI in rats was set up by the improved device of Feeney's weight-dropping model with impact 600 g·cm.Sham-operated rats were anesthetized and had their scalps cut and sutured,but were not submitted to craniectomy or treatment.The intraperitoneal injection of XNJ was performed with 10 mL·kg~(-1)·d~(-1) after the models were set up in the therapy group.TI3I group received only normal saline.
3.Measures
3.1 Determination of brain water content
Edema was measured at 1,3,5 and 7d after surgery.Briefly,samples from the peri-contusion area were taken and weighed.Then they were placed in an oven and dried at 110℃for 24 h.After drying,all weighing was done on the same balance.The percent water was calculated by the following equation:%Water =[(wet sample weight-dry sample weight)/(wet sample weight)]×100%
3.2 Determination of BBB permeability
2%Evans blue(EB) dye(3mL·kg~(-1)) was injected into the femoral vein in one hour before decapitation.The chest was opened 60 minutes later and the heart was perfused with 500 mL saline at 37℃through the left ventricle until the colorless perfusion fluid was obtained from the right atrium.After decapitation,tissue samples about 0.2g were obtained and weighed.Then,the samples were placed in 5ml formamide solution and incubated for 24 hours at 45℃.The optical density(OD) of the EB formamide solution was determined by spectrophotometry at 635 nm,then get the content of EB(μg) according to the standard curve.BBB permeability was expressed as EB per gram of tissue(μg·g~(-1)).
3.3 Immunohistochemistry for AQP4 protein expression
Brain tissues were fixed in 4%formaldehyde for 24 - 48 h and then embedded in paraffin.Sample sections were cut and stained with hematoxylin and eosin.Adjacent sections were incubated with a primary antibody against AQP4(1:300) for 1 d at 4℃. Sections were sequentially incubated with anti-goat IgG biotynylated secondary antibodies and avidin biotin complex.Sections were finally visualized with 0.01% diaminobenzidine tetrahydrochloride.To test the specificity of the immunohistochemical reaction,control sections were treated with normal goat serum instead of the primary antibody.Nuclei were counterstained by hematoxylin.Samples were photographed and measured by image analysis software.
3.4 Real-time quantitative RT-PCR for AQP4 mRNA expression
Total RNA was isolated from brain with Trizol reagent according to the manufacture's protocol.The cDNA were amplified with primers based on the sequences of rat AQP4 cDNA.The downstream primer for rat AQP4 was 5′AAC CGT GGT GAC TCC CAA TCC 3′,and the up stream primer was 5′GGT CCT CAT CTC CCT CTG CTT 3′(270 bp).The downstream primer for humanβ-actin cDNA was5′TAG AAG CAT TTG CGG TGC 3′and the upstream primer was 5′TGT GAT GGT GGG TAT GGG 3′(241bp).cDNA samples were amplified for 40 cycles(denaturation at 95℃for 15 s,annealing at 58℃for 15 s and extension at 72℃for 30s).The ABI7500 system software monitored the changes in fluorescence of SYBR Green I dye in every cycle.The threshold cycle(Ct) value for each reaction,reflecting the amount of PCR needed to identify a target gene,and the relative level of AQP4 for each sample were calculated as described.Briefly,β-actin was used for the normalization of the quantity of RNA used.Its Ct value was then subtracted from that of the AQP4 gene to obtain a△Ct value.The difference(△△Ct) between the△Ct values of the samples for the gene target and the△Ct value of the calibrator was determined.The sham-operation group was chosen for calibrators.The relative quantitative value was expressed as 2~(-△△Ct), representing the amount of AQP4 expression relative to the calibrators.
4.Statistical analysis
All data were expressed as mean±SD.Statistical analysis was carried out with t test by using SPSS 11.5 statistical package.P<0.05 was considered significant.
Results
1.Effect of XNJ on brain water content
In TBI group,cerebral edema developed and peaked from 1d to 3d,which were much higher than sham operation group(P<0.05) and decreased significantly by 5 days. At 7d,it was still higher than sham operation group(P<0.05).XNJ reduced the level of cerebral edema at 1,3,5d as compared to those in TBI group(P<0.05).At 7d,it was very close to sham operation group(P＞0.05).
2.Effect of XNJ on BBB permeability
BBB permeability in TBI group increased and peaked from 1d,and then declined gradually at every measure times,which were much higher than sham operation group(P＜0.05).XNJ injections reduced the leak of BBB as compared to those in TBI group(P＜0.05).But they were still higher than sham operation group(P＜0.05).
3.BBB permeability was coincident with brain edema development with good correlation (r=0.898,P＜0.01).
4.Effect of XNJ on the expressions of AQP4 mRNA and protein
The expressions of AQP4 mRNA and protein strongly detected at 1-3d after TBI which were higher than sham operation group(P＜0.05),then decreased at 5d.XNJ significantly increased AQP4 expressions at 1d-3d post-injury in the brain tissue and much higher than TBI(P＜0.05).
Conclusion
XNJ could alleviate cerebral edema following traumatic brain injury via reducing blood-brain barrier damage and the up-regulation of AQP4 expressions.
通过建立sD大鼠自由落体脑创伤模型,观察腹腔注射XNJ对大鼠急性颅脑创伤后脑水肿、BBB的损害及AQP4的影响,探讨其脑保护作用及作用机制。
研究方法
1.实验动物
清洁级、健康、成熟sD雄性大鼠108只,体重300～350g。随机分为假手术组、脑外伤组和XNJ治疗组。后两组再分为伤后1、3、5、7d等4个亚组,每亚组12只。
2.实验方法
采用参照Feeney'S自由落体模型设计并适当改进的打击装置。用20 g重的击锤从30 cm高处自由坠落冲击撞杆,冲击力为600 g·cm,造成脑挫裂伤,制作大鼠脑创伤模型。XNJ治疗组在大鼠脑外伤模型做好后10 min内腹腔内注射XNJ 10mL·kg~(-1),每天一次。脑外伤组则腹腔内给予等量0.9%氯化钠溶液。假手术组仅行开颅术,不造成脑损伤。
3.标本采集和检测
3.1脑组织含水量检测将大鼠伤后第1、3、5、7d分别断头处死后,立即取出伤侧大鼠脑组织,110℃恒温箱烘烤24 h至恒重。用干湿质量法计算脑组织含水量。计算公式为:脑组织含水量(%)=(湿质量-干质量)/湿质量×100%。假手术组术后24 h做同样处理。
3.2 BBB定量测定采用EB比色法。各组于处死前1 h通过股静脉注射2%EB(3mL·kg~(-1))。取脑组织前,快速开胸暴露心脏,迅速穿刺左心室后,剪右心房一小口,用37℃生理盐水500 mL灌注,至右心房流出清亮液体。取伤灶周边脑组织约0.2 g,加人5 mL甲酰胺,45℃水浴24 h,1500 r·min~(-1)离心10 min,取上清液用HP8453型分光光度计(λ=635 nm)测其光密度值。用倍比稀释法制作标准曲线,再根据标准曲线求得脑组织EB含量,结果以μg·g~(-1)(脑组织)表示。
3.3 AQP4免疫组化染色各组大鼠在预定时间点在伤灶区周围取脑组织,4%甲醛固定24-48 h,石蜡包埋,然后切片行HE染色和免疫组化检查。以细胞膜呈棕黄色或褐色为AQP4阳性染色。应用NIKON彩色病理图文分析系统进行图像分析。
3.4脑组织AQP4 mRNA的实时荧光RT-PCR测定分别取各组不同时间点的损伤周围脑组织进行RT-PCR分析,按Trizol试剂说明提取组织总RNA,逆转录为cDNA,以逆转录所得eDNA为模板进行PCR扩增。AQP4引物:上游5′GGT CCT CAT CTC CCTCTG CTT 3′,下游5′AAC CGT GGT GAC TCC CAA TCC3′,扩增片段长270 bp;β-肌动蛋白引物:上游5′TGT GAT GGT GGG TAT GGG 3′,下游5′TAG AAG CATTTG CGG TGC 3′,扩增片段长度为241 bp。反应体系50 uL,PCR条件:95℃预变性3 min;再95℃变性15 s,58℃退火15 s,72℃延伸30 s,共40个循环。求得各基因的Ct值。采用2~(-ΔΔCt)方法,即以管家基因β-肌动蛋白的Ct值标准化目的基因Ct值,得到各时点目的基因初始模板相对量,再计算各时点目的基因初始模板相对量与假手术组目的基因初始模板相对量的比值,分析各时点目的基因AQP4mRNA表达相对量的变化。
4.统计学处理
采用SPSS11.5统计软件包进行处理,资料以均数±标准差((?)±s)表示,采用单因素方差分析方法(One-Way ANOVA);P＜0.05为差异有统计学意义。
研究结果
1.各组大鼠脑含水量比较
脑外伤组脑组织含水量伤后1d达高峰,并持续至第3d,第5d开始下降,第7天仍明显高于假手术组(P＜0.05)。XNJ组脑含水量也在第1d达高峰,后逐渐下降,但均明显低于脑外伤组(P＜0.05);并于第7d降至接近正常,与假手术组比较无显著性差异(P＞0.05)。
2.各组大鼠脑组织EB含量比较
脑外伤组EB含量伤后1d达高峰,后逐渐下降,各时点均明显高于假手术组(P＜0.05)。XNJ组EB含量伤后1d达高峰,也逐渐下降,各时点均明显低于脑外伤组(P＜0.05),但均不同程度高于假手术组。
3.脑含水量与EB的相关分析
脑含水量与EB的变化规律相同,相关分析得,r=0.898,P＜0.01,表明二者具有正性相关。
4.AQP4免疫组化染色和AQP4 mRNA
光学显微镜下观察,以细胞膜呈棕黄色或褐色为AQP4阳性染色。脑外伤组伤后1d AQP4免疫染色变深,它的mRNA水平上调,并持续至第3d,明显高于假手术组(P＜0.05);在第5d开始下降,与假手术组比较无显著性差异(P＞0.05)。XNJ治疗后1、3 d的AQP4和AQP4 mRNA较脑外伤组明显上调(P＜0.05)。
结论
XNJ可以减轻创伤性脑水肿,可能与其减轻BBB破坏,上调AQP4表达有关。
Objective
In order to clarify the protective effects and the mechanism of XNJ,TBI rat models were established,brain water content,BBB permeability,AQP4 and AQP4 mRNA were tested. Methods
1.Subjects
108 male Sprague-Dawley rats approximately 300-350g,were used as subjects and randomly assigned to three groups:sham-operation,TBI and XNJ group.The later two groups were divided into 4 subgroups at 1,3,5 and 7d.
2.Models
TBI in rats was set up by the improved device of Feeney's weight-dropping model with impact 600 g·cm.Sham-operated rats were anesthetized and had their scalps cut and sutured,but were not submitted to craniectomy or treatment.The intraperitoneal injection of XNJ was performed with 10 mL·kg~(-1)·d~(-1) after the models were set up in the therapy group.TI3I group received only normal saline.
3.Measures
3.1 Determination of brain water content
Edema was measured at 1,3,5 and 7d after surgery.Briefly,samples from the peri-contusion area were taken and weighed.Then they were placed in an oven and dried at 110℃for 24 h.After drying,all weighing was done on the same balance.The percent water was calculated by the following equation:%Water =[(wet sample weight-dry sample weight)/(wet sample weight)]×100%
3.2 Determination of BBB permeability
2%Evans blue(EB) dye(3mL·kg~(-1)) was injected into the femoral vein in one hour before decapitation.The chest was opened 60 minutes later and the heart was perfused with 500 mL saline at 37℃through the left ventricle until the colorless perfusion fluid was obtained from the right atrium.After decapitation,tissue samples about 0.2g were obtained and weighed.Then,the samples were placed in 5ml formamide solution and incubated for 24 hours at 45℃.The optical density(OD) of the EB formamide solution was determined by spectrophotometry at 635 nm,then get the content of EB(μg) according to the standard curve.BBB permeability was expressed as EB per gram of tissue(μg·g~(-1)).
3.3 Immunohistochemistry for AQP4 protein expression
Brain tissues were fixed in 4%formaldehyde for 24 - 48 h and then embedded in paraffin.Sample sections were cut and stained with hematoxylin and eosin.Adjacent sections were incubated with a primary antibody against AQP4(1:300) for 1 d at 4℃. Sections were sequentially incubated with anti-goat IgG biotynylated secondary antibodies and avidin biotin complex.Sections were finally visualized with 0.01% diaminobenzidine tetrahydrochloride.To test the specificity of the immunohistochemical reaction,control sections were treated with normal goat serum instead of the primary antibody.Nuclei were counterstained by hematoxylin.Samples were photographed and measured by image analysis software.
3.4 Real-time quantitative RT-PCR for AQP4 mRNA expression
Total RNA was isolated from brain with Trizol reagent according to the manufacture's protocol.The cDNA were amplified with primers based on the sequences of rat AQP4 cDNA.The downstream primer for rat AQP4 was 5′AAC CGT GGT GAC TCC CAA TCC 3′,and the up stream primer was 5′GGT CCT CAT CTC CCT CTG CTT 3′(270 bp).The downstream primer for humanβ-actin cDNA was5′TAG AAG CAT TTG CGG TGC 3′and the upstream primer was 5′TGT GAT GGT GGG TAT GGG 3′(241bp).cDNA samples were amplified for 40 cycles(denaturation at 95℃for 15 s,annealing at 58℃for 15 s and extension at 72℃for 30s).The ABI7500 system software monitored the changes in fluorescence of SYBR Green I dye in every cycle.The threshold cycle(Ct) value for each reaction,reflecting the amount of PCR needed to identify a target gene,and the relative level of AQP4 for each sample were calculated as described.Briefly,β-actin was used for the normalization of the quantity of RNA used.Its Ct value was then subtracted from that of the AQP4 gene to obtain a△Ct value.The difference(△△Ct) between the△Ct values of the samples for the gene target and the△Ct value of the calibrator was determined.The sham-operation group was chosen for calibrators.The relative quantitative value was expressed as 2~(-△△Ct), representing the amount of AQP4 expression relative to the calibrators.
4.Statistical analysis
All data were expressed as mean±SD.Statistical analysis was carried out with t test by using SPSS 11.5 statistical package.P<0.05 was considered significant.
Results
1.Effect of XNJ on brain water content
In TBI group,cerebral edema developed and peaked from 1d to 3d,which were much higher than sham operation group(P<0.05) and decreased significantly by 5 days. At 7d,it was still higher than sham operation group(P<0.05).XNJ reduced the level of cerebral edema at 1,3,5d as compared to those in TBI group(P<0.05).At 7d,it was very close to sham operation group(P＞0.05).
2.Effect of XNJ on BBB permeability
BBB permeability in TBI group increased and peaked from 1d,and then declined gradually at every measure times,which were much higher than sham operation group(P＜0.05).XNJ injections reduced the leak of BBB as compared to those in TBI group(P＜0.05).But they were still higher than sham operation group(P＜0.05).
3.BBB permeability was coincident with brain edema development with good correlation (r=0.898,P＜0.01).
4.Effect of XNJ on the expressions of AQP4 mRNA and protein
The expressions of AQP4 mRNA and protein strongly detected at 1-3d after TBI which were higher than sham operation group(P＜0.05),then decreased at 5d.XNJ significantly increased AQP4 expressions at 1d-3d post-injury in the brain tissue and much higher than TBI(P＜0.05).
Conclusion
XNJ could alleviate cerebral edema following traumatic brain injury via reducing blood-brain barrier damage and the up-regulation of AQP4 expressions.
引文
[1]JONATHAN W,GRAHAM M.Management of severe head injury [J].ContempNeurosurg,1996,18:1-5.
[2]易声禹,只达石主编.颅脑损伤诊治.北京:人民卫生出版社,2000.3.
[3]CAREY ME,SARNA GS,FARRELL JB.Brain edema following an experimental missile wound to the brain[J].Neurotrauma,1999,24(7):13-20.
[4]OEHMICHEN M,GEHL HB,MEISSNER C,et al.Forensic pathological aspects of postmortem imaging of gunshot injury to the head:documentation and biometric data[J].Acta Neuropathol,2002,105(6):570-580.
[5]SHAPIRA Y,SHOHAMI E,SIDI A,et al.Experimental closed head injury in rats:mechanical,pathophysiologic,and neurologic properties[J].Crit Care Med,2004,16(4):258-265
[6]黄卫东,苏伟,徐妙等.急性颅脑损伤后精氨酸加压素动态变化的临床研究[J].中华创伤杂志;2006,22(3):190-193.
[7]TONI D,CHAMORRO A,KASTE M,et al.Acute treatment of ischemic stroke.European Stroke Initiative[J].Cerebrovasc Dis,2004,17(suppl 2):30.
[8]KAPPELLE IJ,VAN DER WORP HB.Treatment or prevention of complications of acute ischemic stroke[J].CurrNeurol Neurosci Rep,2004,4(1):36.
[9]XU Miao,SU Wei,HUANG Wei-dong,et al.Effect of AVP on brain edema following traumatic brain injury[J].Chinese Journal ofTraumatology,2007,10(2):90-93
[10]KLATZO I.Evolution of understanding concepts of brain edema.In:The 9th international seminar on brain edema(Abstract book).1993:1.
[11]PRESTON GM,CARROLL TP,GUGGINO WB,et al.Appearance of water channels in Xenopusoocytes expressing red cell CHIP28 protein[J].Science,1992,256(6):385-387.
[12]VERKMAN AS.Physiological importance of aquaporin water channels[J].Ann Med,2002,34(3):192-200.
[13]徐妙,黄卫东,徐秋萍.水通道蛋白-4与创伤性脑水肿的研究进展[J].中华创伤杂志,2007,23(4):314-316.
[14]MANLEY GT,FUJIMURA M,MA T,et al.Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke[J].Nat Med,2000,6(6):159-163.
[15]PAPADOPOULOS MC,MANLEY GT,KRISHNA S,et al.Aquaporin-4 facilitates reabsorption of excess fluid in vasogenic brain edema[J].FASEB J,2004,18(4):1291-1293.
[16]FODA MA,MARAROU A.A new modal of brain injury in rats[J].J Neurosurg,1994,80(6):301-313.
[17]MAXWELL WL,IRVINE A,ADAMS JH,et al.Response of cerebral microvas culature to brain injury[J].J Pathol,1998,15(5):327-335.
[18]IKEDA Y,WANG M,NAKAZAWA S.Simple quantitative evaluation of blood-brain barrier disruption in vasogenic brain edema.Aeta Neuroehir,1994,60(7):119
[19]VIZUETE ML,VENERO JL.,VARGAS C,et al.Differential upregulation of aquaporin-4 mRNA expression in reactive astrocytes after brain injury:potential role in brain edema[J].Neurobiol Dis,2004,6(2):245-258.
[20]NICO B,FRIGCRI A,NIECHIA GP,et al.Role of aquaporin-4 water channel in the development and integrity of the blood-brain barrier[J].Cell Sci 2001,114(7):1297-1307.
[21]王宁生,梁美蓉,刘启德,等.冰片“佐使则有功”之实验研究[J].中医杂志,1994,35(1):46-48.
[22]张路啥,向金莲,程睿,等.醒脑静注射液的药效学研究[J].华西药学杂志,2001,16(6):429-431.
[23]沈思钰,傅晓东,费震宇,等.醒脑静注射液的临床应用进展[J].中国中医急症,2004,13(1):49.
[24]刘宇清.醒脑静注射液治疗急性颅脑损伤意识障碍60例[J].福建中医药,2005,36(5):11-12.
[25]顾斌芳,潘仁龙,金文秀,等.醒脑静对颅脑损伤患者生存质量的影响[J].现代中西医结合杂志,2002,11(8):686-687.
[26]钟恺立,钟兴美,付虹,等.醒脑静注射液对脑损伤后血液流变学改变的治疗作用[J].成都中医药大学学报,2006,29(3):20-21.
[27]FEENEY DM,BOYESON MG,LINN RT,et al.Response to cortical injury:methodology and local effects of contusions in the rat Brain[J].Brain Res,1981,211(1):67-77.
[28]申洪,陆药丹.免疫组织化学染色的定量方法研究[J].生物医学工程学杂志,1993,10(3):281-284
[29]KENNETH J L,THOMAS D S.Analysis of relative gene expression data using real-time quantitative PCR and the method[J].Methods,2002,25,402-408.
[30]LEKER RR,SHOHANTI E,CONSTAN TINI S.Experimental models of head.Trauma [J].Acta Neuroehir Suppl,2002,83(5):49-54.
[31]DIXON CE,LYETH BG,POVLISHOCK YR,et al.A fluid percussion model of experimental brain injury in the rats[J].Neurosurg,1987,67(1):110-119.
[32]李力仙,王天佑,钟震宇,等.自由落体和液压致大鼠脑损伤的对比研究[J].中华实验外科杂志,1999,16(5):447-448.
[33]KIMELBERG HK.Current concepts of brain edema.Review of laboratory investigations[J].Neurosurg,1995,83(4):1051-1059.
[34]BADAUT,J,LASBENNES F,MAGISTRETTI PJ,et al.Aquaporins in brain:distribution,physiology,and pathophysiology[J].Cereb.Blood Flow Metab,2002, 22(6):367 378.
[35]STROOP R,THOMALE UW,PAUSER S,et al.Magnetic resonance imaging studies with cluster algorithm for characterization of brain edema after controlled cortical impact injury(CCII)[J].Acta Neurochir,Suppl,1998,71(3):303-305.
[36]UNTERBERG,AW,STROOP R,THOMALE UW,et al.CharaCterisation of brain edema following controlled cortical impaCt injury" in rats[J].Acta Neurochir,Suppl,1997,70(5):106-108.
[37]UNTERBERG AW,STOVER J,KRESS B,et al.Edema and brain trauma[J].Neuroscience,2004,129(4):1021-1029.
[38]BETA AL,IANNOTTI F,HOFF JT.Brain edema:a classification based on blood-brain barrier integrity[J].Cerebrovasc Brain Metab Rev 1989,1(6):133-154.
[39]KLATZO I.Evolution of brain edema concepts[J].Acta Neurochir,Suppl 1994,60(4):3-6.
[40]BEAUMONT A,MARMAROU A,FATOUROS P,et al.Secondary insults worsen blood brain barrier dysfunction assessed by MRI in cerebral contusion[J].Acta Neurochir,Suppl,2002,81(2):217-219.
[41]NEAL CJ,LEE EY,GYORGY A,et al.Effect of penetrating brain injury on aquaporin-4 expression using a rat model[J].Neurotrauma,2007,24(10):1609-17
[42]VERKMAN AS.Applications of aquaporin inhibitors[J].Drug News Perspect,2003, 14(7):412-420.
[43]NEELY JD,NIELSON S,AGRE P,et al.Heterotetrameric composition of aquaporln-4 water channels[J].Biochemistry,1999,38(34):11156-11161
[44]SUN MC,HONEY CR,BERK C,et al.Regulation of aquaporin-4 in a traumatic brain injury model in rats[J].Neurosurg,2003,98(6):565-569.
[45]KIENING KL,LANDEGHEM FK,SCHREIBER S,et al.Decreased hemispheric Aquaporin-4 is linked to evolving brain edema following controlled cortical impact injury in rats[J].Neurosci Lett,2002,324(6):105-108.
[46]KE C,POON WS,NG HK,et al.Impact of experimental acute hyponatremia on seer traumatic brain injury in rats:influences on injuries,permeability of blood-brain barrier,ultrastructural features,and aquaporin-4 expression[J].Exp Neurol,2002,178(5):194-206.
[47]WANG K,BEKAR K,FURBER GH,et al.Vimentin-expressing proximal reactive astrocytes correlate with migration rather than proliferation following focal brain injury [J].Brain Res.,2004,1024(4):193-202.
[48]MARMAUROU G,HOCHWALD T,NAKAMURA D,et al.Brain edema resolution by CSF pathways and brain vasculature in cats[J].Am J Physiol,1994,267(2):514-520.
[49]REULEN HJ,GRAHAM R,SPATZ M,et al.Role of pressure gradients and bulk flow in dynamics of vasogenic brain edema[J].Neurosurg,1977,46(6):24-35.
[50]BRIGHTMAN MW.The brain's interstitial clefts and their glial walls[J].Neurocytol,2002,31(4):595-603.
[51]NAGELHUS EA,HORIO Y,INANOBE A,et al.Immunogold evidence suggests that coupling of K+ siphoning and water transport in rat retinal Muller cells is mediated by a coenrichment of Kir4.1 and AQP4 in specific membrane domains[J].Glia,1999,26(5):47-54.
[52]PAULSON OB,NEWMAN EA.Does the release of potassium from astrocyte end feet regulate cerebral blood flow?[J].Science,1997,237(5):896-898.
[53]EID T,LEE TS,THOMAS ML,et al.Loss of perivascular aquaporin 4 may underlie deficient water and K+ homeostasis in the human epileptogenic hippocampus[J].Proc.Natl.Acad.Sci.U.S.A.2005,102(9):1193-1198.
[54]李瑾翡,王宁生,陆惠文,等.麝香酮药代动力学研究[J].中药新药与临床药理,2000,11(4):208.
[55]蒋振亚,李常度,周东,等.麝香对大鼠实验性脑缺血神经元损伤的保护作用[J].中国中医药科技,2001,8(2):97.
[56]赵保胜,宓穗卿,张银卿.冰片对大鼠脑微血管ICAM-1表达的影响[J].中药新药与临床药理,2001,12(2):88.
[57]IKEDA Y,WANG M,NAKAZAWA S.Simple quantitative evaluation ofblood-brain barrier disruption in vasogenic brain edema[J].Acta Neurochir,1994,60(Suppl):119-120.
[58]IKEDA Y,WANG M,NAKAZAWA S.Early changes of blood-brain barrier and superoxides cavenging activity in rat cryogenic brain injury[J].Acta Neurochir,1994,60(Suppl):136-138.
[59]戴永建,戢翰升.醒脑静注射液对大鼠脑损伤后细胞凋亡及相关蛋白表达的影响[J].中国临床神经外科杂志,2006,11(9):551-553.
[60]张红,朱志安,陈鑫,等.醒脑静注射液对大鼠脑外伤后脑组织c-fos基因表达的影响[J].药学服务与研究,2002,2(4):246-248.
[61]吴玉生,姜立平,王占奎,等.醒脑静注射液对急性大灶性脑梗塞患者血清肿瘤坏死因子、可溶性血管细胞黏附分子-1的影响[J].中药药理与临床,1999,15(5):202-203
[62]吴玉生,王钊,姜立平.醒脑静注射液对急性脑梗死患者单核细胞CD~(+54)分子表达的影响[J].中国中西医结合急救杂志,1999,6(5):292-293.
[63]陈寿权,王万铁,王明山,等.醒脑静对家兔脑缺血再灌注时TNF、IL-1、IL-6水平及脑超微结构影响的实验研究[J].中国急救医医学,2000,20(11):654-655.
[64]潘光强,易兴阳.醒脑静注射液治疗急性脑梗死临床观察[J].河北中医,2002,24(10):787-78.
[65]孙蓉媚.醒脑静注射液治疗脑梗塞132例—附虚实两证疗效分析[J].浙江中医杂 志,2002,37(9):409-410
[66]马承泰,蒋艳霞,王左.醒脑静生脉混合液对凝血酶损伤星形胶质细胞的保护作用[J].中国中西医结合急救杂志,2003,10(1):46-47.
[67]罗海燕,万文成,陈洁文,等.醒脑静对脑缺血大鼠大脑皮质超微结构的保护作用[J].中医药研究,2000,16(4):37-38.
[68]ZHAO J,MOORE AN,CLIFTON GL,et al.Sulforaphane enhances aquaporin-4expression and decreases cerebral edema following traumatic brain injury[J].J Neurosci Res,2005,82(3):499-506.
[69]NICO B,FRIGCRI A,NIECHIA GP,et al Role of aquaporin-4 water channel in the development and integrity of the blood brain barrier[J].J Cell Sci,2001,114(7):1297-1307.
[70]杨明,罗培,孟现民,等.醒脑静注射液的一般药理学研究[J].华西药学杂志,2005,20(3):215-217.
[71]BALDWIN SA,FUGACCIA I,BROWN DR,et al.Blood-brain barrier breach following cortical conmsionintherat[J].Neurosurg,1996,85(4):476-481.
[72]FUKUDA K,TANNO H,OKIMURA Y,et al.The blood-brain barrier diruption to circulatting proteins in the early period after fluid percussion brain injury in rots[J],Neurotrauma,1995,12(3):315-324.
[73]郭宗泽,曹春阳,杨毅,等.醒脑静粉针对实验性脑外伤的保护作用[J].沈 阳药科大学学报,2006,23(6):384-387.
[1]PRESTON GM,CARROLL TP,GUGGINO WB,et al.Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein[J].Science,1992,256(6):385-387.
[2]VERKMAN AS.Physiological importance of aquaporin water channels[J].Ann Med,2002,34(4):192-200.
[3]YANG B.The human aquaporin gene family(review)[J].Current Genomics,2000,1(9):91-102.
[4]LU M,LEE MD,SMITH B.The human AQP4 gene:definition of the locus encoding two water channel polypeptide[J].Proc Natl Acad Sci USA,1996,93(11):10908-10912.
[5]VERKMAN AS.Applications of aquaporin inhibitors[J].Drug News Perspect,2003,14(2):412-420.
[6]YANG B,MA T,VERKMAN AS.cDNA coning,gene organization and chromosomal localization of a human mercurial-insensitive water channel:evidence for distinct transcriptional units[J].J Biol Chem,1995,270(1):22907-22913.
[7]VENERO JL,VIZUETE ML,MACHADO A,et al.Aquaporins in the central nervous system[J].Prog Neurobiol,2001,63(7):121-130.
[8]YAMAOTO N,YONEDA K,ASAI K,et al.Alteration in the expression of AQP family in cultured rat astrocytes hypoxia and reoxygenation[J].Brain Res Mol Brain Res,2001,90(6):21-24.
[9]KOBAYASHI H,MINAMI S,ITOH S,et al.Aquaporin subtypes in rat cerebral microvessels[J].Neurosic Lett,2001,297(4):163-166.
[10]YONEDA K,YAMAMOTO N,ASAI K,et al.Regulation of aquaporin-4 expression in astrocytes[J].Brain Res Mol Brain Res,2001,89(6):94-102.
[11]AMIRY-MOGHADDAM M,OTSUKS T,HURN PD,et al.An alpha-syntrophin-dependent pool of AQP4 in astroglial end-feet confers bidirectional water flow between blood and brain[J].Proc Natl Acad Sci USA,2003,100(8):2106-2111.
[12]PATIL RV,HAN Z.Regulation of water-channel aCtivity of aquaporins by arginine vasopression and atrial natriuretic peptide[J].Biochem Biophys Res Commun,1997,238(7):392-339.
[13]HASEGAWA H,MA T,SKACH W,et al.Molecular cloning of a mercurial-insensitive water channel expressed inselected water-transporting tissues [J].Biol Chem,1994,269(8):5497-5500.
[14]ARIMA H,YAMAMOTO N,SOBUE K,et al.Hyperosmolar mannitol simulates expression of aquaporin 4 and 9 through a p38 mitogen -activated protein kinase-dependent pathway in rat astrocytes[J].J Biol Claem,2003,278(8):44525-44534.
[15]MANLEY GT,FUIMURA M,MA T,et al.Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke[J].Nat Med,2000,6(9):159-163.
[16]AMORINI AM,DUNBAR JG,MARMAROU A.Modulation of aquaporin-4water transport in a model of TBI[J].ACta Neurochir,2003,86(Suppl):261-263.
[17]KE C,POON WS,NG HK,et al.ImpaCt of experimental acute hyponatremia on seer traumatic brain injury in rats:influences on inuries,permeability of blood-brain barrier,ultrastructural features,and aquaporin-4 expression[J].Exp Neurol,2002,178(7):194-206.
[18]KIENING KL,LANDEGHEM FK,SCHREIBER S,et al.Decreased hemispheric Aquaporin-4 is linked to evolving brain edema following controlled cortical impact injury in rats[J].Neurosci Lett,2002,324(8):105-108.
[19]SUN MC,HONEY CR,BERK C,et al.Regulation of aquaporin-4 in a traumatic brain injury model in rats[J].J Neurosurg,2003,98(4):565-569.
[20]SAADOUN S,PAPADOPOULOS MC,KRISHNA S.Water transport becomes uncoupled from k+ siphoning in brain contusion,bacterial meningitis,and brain tumours:immunohistochemical case review[J].J Clin Pathol,2003,56(8):972-975.
[21]BADAUT J,BRUNNET JE,GROLLIMUND L,et al.Aquaporin 1 and aquaprin 4 experession in human brain after subarachnoid hemorrhage and in peritumoral tissue[J].Acta Neurochir,2003,86(Suppl):495-498.
[22]HU H,YAO HT,ZHANG WP,et al.Increased expression of aquaporin-4 in human traumatic brain injury and brain tumors[J].J Zhejiang Univ Sci B,2005,6(5):33-37.
[23]PAPADOPOULOS MC,MANLEY GT,KRISHNA S,et al.Aquaporin-4 facilitates reabsorption of excess luid in vasogenic brain edema[J].FASEB J,2004,18(6):1291-1293.
[24]KE C,POON WS,NG HK,et al.Heterogeneous responses of aquaporin-4 in oedema formation in a replicated severe traumatic brain injury model in rats [J].Neurosci Lett,2001,301(3):21-24.
[25]ZHAO J,MOORE AN,CLIFTON GL,et al.Sulforaphane enhances aquaporin-4expression and decreases cerebral edema following traumatic brain injury[J].J Neurosci Res,2005,82(7):499-506.
[26]GUO Q,SAVEED I,BARONNE LM,et al.Progesterone administration modulates AQP4 expression and edema after traumatic brain injury in mate rats[J].Exp Nenrol,2006,198(2):469-478.
[27]NIERMANN H,AMIRY-MOGHADDAM M,HOLTHOFF K,et al.A novel role of vasopressin in the brain:modulation of activity-dependent water flux in the neocortex[J].J Neurosei,2001,21(9):3045-3051.
[28]VENERO JI,VIZUETE ML.Detailed localization of aquaporin-4 messenger RNA in the CNS:preferential expression in periventricular organs[J].Neuroscience,1999,94(7):239-250.
[2]易声禹,只达石主编.颅脑损伤诊治.北京:人民卫生出版社,2000.3.
[3]CAREY ME,SARNA GS,FARRELL JB.Brain edema following an experimental missile wound to the brain[J].Neurotrauma,1999,24(7):13-20.
[4]OEHMICHEN M,GEHL HB,MEISSNER C,et al.Forensic pathological aspects of postmortem imaging of gunshot injury to the head:documentation and biometric data[J].Acta Neuropathol,2002,105(6):570-580.
[5]SHAPIRA Y,SHOHAMI E,SIDI A,et al.Experimental closed head injury in rats:mechanical,pathophysiologic,and neurologic properties[J].Crit Care Med,2004,16(4):258-265
[6]黄卫东,苏伟,徐妙等.急性颅脑损伤后精氨酸加压素动态变化的临床研究[J].中华创伤杂志;2006,22(3):190-193.
[7]TONI D,CHAMORRO A,KASTE M,et al.Acute treatment of ischemic stroke.European Stroke Initiative[J].Cerebrovasc Dis,2004,17(suppl 2):30.
[8]KAPPELLE IJ,VAN DER WORP HB.Treatment or prevention of complications of acute ischemic stroke[J].CurrNeurol Neurosci Rep,2004,4(1):36.
[9]XU Miao,SU Wei,HUANG Wei-dong,et al.Effect of AVP on brain edema following traumatic brain injury[J].Chinese Journal ofTraumatology,2007,10(2):90-93
[10]KLATZO I.Evolution of understanding concepts of brain edema.In:The 9th international seminar on brain edema(Abstract book).1993:1.
[11]PRESTON GM,CARROLL TP,GUGGINO WB,et al.Appearance of water channels in Xenopusoocytes expressing red cell CHIP28 protein[J].Science,1992,256(6):385-387.
[12]VERKMAN AS.Physiological importance of aquaporin water channels[J].Ann Med,2002,34(3):192-200.
[13]徐妙,黄卫东,徐秋萍.水通道蛋白-4与创伤性脑水肿的研究进展[J].中华创伤杂志,2007,23(4):314-316.
[14]MANLEY GT,FUJIMURA M,MA T,et al.Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke[J].Nat Med,2000,6(6):159-163.
[15]PAPADOPOULOS MC,MANLEY GT,KRISHNA S,et al.Aquaporin-4 facilitates reabsorption of excess fluid in vasogenic brain edema[J].FASEB J,2004,18(4):1291-1293.
[16]FODA MA,MARAROU A.A new modal of brain injury in rats[J].J Neurosurg,1994,80(6):301-313.
[17]MAXWELL WL,IRVINE A,ADAMS JH,et al.Response of cerebral microvas culature to brain injury[J].J Pathol,1998,15(5):327-335.
[18]IKEDA Y,WANG M,NAKAZAWA S.Simple quantitative evaluation of blood-brain barrier disruption in vasogenic brain edema.Aeta Neuroehir,1994,60(7):119
[19]VIZUETE ML,VENERO JL.,VARGAS C,et al.Differential upregulation of aquaporin-4 mRNA expression in reactive astrocytes after brain injury:potential role in brain edema[J].Neurobiol Dis,2004,6(2):245-258.
[20]NICO B,FRIGCRI A,NIECHIA GP,et al.Role of aquaporin-4 water channel in the development and integrity of the blood-brain barrier[J].Cell Sci 2001,114(7):1297-1307.
[21]王宁生,梁美蓉,刘启德,等.冰片“佐使则有功”之实验研究[J].中医杂志,1994,35(1):46-48.
[22]张路啥,向金莲,程睿,等.醒脑静注射液的药效学研究[J].华西药学杂志,2001,16(6):429-431.
[23]沈思钰,傅晓东,费震宇,等.醒脑静注射液的临床应用进展[J].中国中医急症,2004,13(1):49.
[24]刘宇清.醒脑静注射液治疗急性颅脑损伤意识障碍60例[J].福建中医药,2005,36(5):11-12.
[25]顾斌芳,潘仁龙,金文秀,等.醒脑静对颅脑损伤患者生存质量的影响[J].现代中西医结合杂志,2002,11(8):686-687.
[26]钟恺立,钟兴美,付虹,等.醒脑静注射液对脑损伤后血液流变学改变的治疗作用[J].成都中医药大学学报,2006,29(3):20-21.
[27]FEENEY DM,BOYESON MG,LINN RT,et al.Response to cortical injury:methodology and local effects of contusions in the rat Brain[J].Brain Res,1981,211(1):67-77.
[28]申洪,陆药丹.免疫组织化学染色的定量方法研究[J].生物医学工程学杂志,1993,10(3):281-284
[29]KENNETH J L,THOMAS D S.Analysis of relative gene expression data using real-time quantitative PCR and the method[J].Methods,2002,25,402-408.
[30]LEKER RR,SHOHANTI E,CONSTAN TINI S.Experimental models of head.Trauma [J].Acta Neuroehir Suppl,2002,83(5):49-54.
[31]DIXON CE,LYETH BG,POVLISHOCK YR,et al.A fluid percussion model of experimental brain injury in the rats[J].Neurosurg,1987,67(1):110-119.
[32]李力仙,王天佑,钟震宇,等.自由落体和液压致大鼠脑损伤的对比研究[J].中华实验外科杂志,1999,16(5):447-448.
[33]KIMELBERG HK.Current concepts of brain edema.Review of laboratory investigations[J].Neurosurg,1995,83(4):1051-1059.
[34]BADAUT,J,LASBENNES F,MAGISTRETTI PJ,et al.Aquaporins in brain:distribution,physiology,and pathophysiology[J].Cereb.Blood Flow Metab,2002, 22(6):367 378.
[35]STROOP R,THOMALE UW,PAUSER S,et al.Magnetic resonance imaging studies with cluster algorithm for characterization of brain edema after controlled cortical impact injury(CCII)[J].Acta Neurochir,Suppl,1998,71(3):303-305.
[36]UNTERBERG,AW,STROOP R,THOMALE UW,et al.CharaCterisation of brain edema following controlled cortical impaCt injury" in rats[J].Acta Neurochir,Suppl,1997,70(5):106-108.
[37]UNTERBERG AW,STOVER J,KRESS B,et al.Edema and brain trauma[J].Neuroscience,2004,129(4):1021-1029.
[38]BETA AL,IANNOTTI F,HOFF JT.Brain edema:a classification based on blood-brain barrier integrity[J].Cerebrovasc Brain Metab Rev 1989,1(6):133-154.
[39]KLATZO I.Evolution of brain edema concepts[J].Acta Neurochir,Suppl 1994,60(4):3-6.
[40]BEAUMONT A,MARMAROU A,FATOUROS P,et al.Secondary insults worsen blood brain barrier dysfunction assessed by MRI in cerebral contusion[J].Acta Neurochir,Suppl,2002,81(2):217-219.
[41]NEAL CJ,LEE EY,GYORGY A,et al.Effect of penetrating brain injury on aquaporin-4 expression using a rat model[J].Neurotrauma,2007,24(10):1609-17
[42]VERKMAN AS.Applications of aquaporin inhibitors[J].Drug News Perspect,2003, 14(7):412-420.
[43]NEELY JD,NIELSON S,AGRE P,et al.Heterotetrameric composition of aquaporln-4 water channels[J].Biochemistry,1999,38(34):11156-11161
[44]SUN MC,HONEY CR,BERK C,et al.Regulation of aquaporin-4 in a traumatic brain injury model in rats[J].Neurosurg,2003,98(6):565-569.
[45]KIENING KL,LANDEGHEM FK,SCHREIBER S,et al.Decreased hemispheric Aquaporin-4 is linked to evolving brain edema following controlled cortical impact injury in rats[J].Neurosci Lett,2002,324(6):105-108.
[46]KE C,POON WS,NG HK,et al.Impact of experimental acute hyponatremia on seer traumatic brain injury in rats:influences on injuries,permeability of blood-brain barrier,ultrastructural features,and aquaporin-4 expression[J].Exp Neurol,2002,178(5):194-206.
[47]WANG K,BEKAR K,FURBER GH,et al.Vimentin-expressing proximal reactive astrocytes correlate with migration rather than proliferation following focal brain injury [J].Brain Res.,2004,1024(4):193-202.
[48]MARMAUROU G,HOCHWALD T,NAKAMURA D,et al.Brain edema resolution by CSF pathways and brain vasculature in cats[J].Am J Physiol,1994,267(2):514-520.
[49]REULEN HJ,GRAHAM R,SPATZ M,et al.Role of pressure gradients and bulk flow in dynamics of vasogenic brain edema[J].Neurosurg,1977,46(6):24-35.
[50]BRIGHTMAN MW.The brain's interstitial clefts and their glial walls[J].Neurocytol,2002,31(4):595-603.
[51]NAGELHUS EA,HORIO Y,INANOBE A,et al.Immunogold evidence suggests that coupling of K+ siphoning and water transport in rat retinal Muller cells is mediated by a coenrichment of Kir4.1 and AQP4 in specific membrane domains[J].Glia,1999,26(5):47-54.
[52]PAULSON OB,NEWMAN EA.Does the release of potassium from astrocyte end feet regulate cerebral blood flow?[J].Science,1997,237(5):896-898.
[53]EID T,LEE TS,THOMAS ML,et al.Loss of perivascular aquaporin 4 may underlie deficient water and K+ homeostasis in the human epileptogenic hippocampus[J].Proc.Natl.Acad.Sci.U.S.A.2005,102(9):1193-1198.
[54]李瑾翡,王宁生,陆惠文,等.麝香酮药代动力学研究[J].中药新药与临床药理,2000,11(4):208.
[55]蒋振亚,李常度,周东,等.麝香对大鼠实验性脑缺血神经元损伤的保护作用[J].中国中医药科技,2001,8(2):97.
[56]赵保胜,宓穗卿,张银卿.冰片对大鼠脑微血管ICAM-1表达的影响[J].中药新药与临床药理,2001,12(2):88.
[57]IKEDA Y,WANG M,NAKAZAWA S.Simple quantitative evaluation ofblood-brain barrier disruption in vasogenic brain edema[J].Acta Neurochir,1994,60(Suppl):119-120.
[58]IKEDA Y,WANG M,NAKAZAWA S.Early changes of blood-brain barrier and superoxides cavenging activity in rat cryogenic brain injury[J].Acta Neurochir,1994,60(Suppl):136-138.
[59]戴永建,戢翰升.醒脑静注射液对大鼠脑损伤后细胞凋亡及相关蛋白表达的影响[J].中国临床神经外科杂志,2006,11(9):551-553.
[60]张红,朱志安,陈鑫,等.醒脑静注射液对大鼠脑外伤后脑组织c-fos基因表达的影响[J].药学服务与研究,2002,2(4):246-248.
[61]吴玉生,姜立平,王占奎,等.醒脑静注射液对急性大灶性脑梗塞患者血清肿瘤坏死因子、可溶性血管细胞黏附分子-1的影响[J].中药药理与临床,1999,15(5):202-203
[62]吴玉生,王钊,姜立平.醒脑静注射液对急性脑梗死患者单核细胞CD~(+54)分子表达的影响[J].中国中西医结合急救杂志,1999,6(5):292-293.
[63]陈寿权,王万铁,王明山,等.醒脑静对家兔脑缺血再灌注时TNF、IL-1、IL-6水平及脑超微结构影响的实验研究[J].中国急救医医学,2000,20(11):654-655.
[64]潘光强,易兴阳.醒脑静注射液治疗急性脑梗死临床观察[J].河北中医,2002,24(10):787-78.
[65]孙蓉媚.醒脑静注射液治疗脑梗塞132例—附虚实两证疗效分析[J].浙江中医杂 志,2002,37(9):409-410
[66]马承泰,蒋艳霞,王左.醒脑静生脉混合液对凝血酶损伤星形胶质细胞的保护作用[J].中国中西医结合急救杂志,2003,10(1):46-47.
[67]罗海燕,万文成,陈洁文,等.醒脑静对脑缺血大鼠大脑皮质超微结构的保护作用[J].中医药研究,2000,16(4):37-38.
[68]ZHAO J,MOORE AN,CLIFTON GL,et al.Sulforaphane enhances aquaporin-4expression and decreases cerebral edema following traumatic brain injury[J].J Neurosci Res,2005,82(3):499-506.
[69]NICO B,FRIGCRI A,NIECHIA GP,et al Role of aquaporin-4 water channel in the development and integrity of the blood brain barrier[J].J Cell Sci,2001,114(7):1297-1307.
[70]杨明,罗培,孟现民,等.醒脑静注射液的一般药理学研究[J].华西药学杂志,2005,20(3):215-217.
[71]BALDWIN SA,FUGACCIA I,BROWN DR,et al.Blood-brain barrier breach following cortical conmsionintherat[J].Neurosurg,1996,85(4):476-481.
[72]FUKUDA K,TANNO H,OKIMURA Y,et al.The blood-brain barrier diruption to circulatting proteins in the early period after fluid percussion brain injury in rots[J],Neurotrauma,1995,12(3):315-324.
[73]郭宗泽,曹春阳,杨毅,等.醒脑静粉针对实验性脑外伤的保护作用[J].沈 阳药科大学学报,2006,23(6):384-387.
[1]PRESTON GM,CARROLL TP,GUGGINO WB,et al.Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein[J].Science,1992,256(6):385-387.
[2]VERKMAN AS.Physiological importance of aquaporin water channels[J].Ann Med,2002,34(4):192-200.
[3]YANG B.The human aquaporin gene family(review)[J].Current Genomics,2000,1(9):91-102.
[4]LU M,LEE MD,SMITH B.The human AQP4 gene:definition of the locus encoding two water channel polypeptide[J].Proc Natl Acad Sci USA,1996,93(11):10908-10912.
[5]VERKMAN AS.Applications of aquaporin inhibitors[J].Drug News Perspect,2003,14(2):412-420.
[6]YANG B,MA T,VERKMAN AS.cDNA coning,gene organization and chromosomal localization of a human mercurial-insensitive water channel:evidence for distinct transcriptional units[J].J Biol Chem,1995,270(1):22907-22913.
[7]VENERO JL,VIZUETE ML,MACHADO A,et al.Aquaporins in the central nervous system[J].Prog Neurobiol,2001,63(7):121-130.
[8]YAMAOTO N,YONEDA K,ASAI K,et al.Alteration in the expression of AQP family in cultured rat astrocytes hypoxia and reoxygenation[J].Brain Res Mol Brain Res,2001,90(6):21-24.
[9]KOBAYASHI H,MINAMI S,ITOH S,et al.Aquaporin subtypes in rat cerebral microvessels[J].Neurosic Lett,2001,297(4):163-166.
[10]YONEDA K,YAMAMOTO N,ASAI K,et al.Regulation of aquaporin-4 expression in astrocytes[J].Brain Res Mol Brain Res,2001,89(6):94-102.
[11]AMIRY-MOGHADDAM M,OTSUKS T,HURN PD,et al.An alpha-syntrophin-dependent pool of AQP4 in astroglial end-feet confers bidirectional water flow between blood and brain[J].Proc Natl Acad Sci USA,2003,100(8):2106-2111.
[12]PATIL RV,HAN Z.Regulation of water-channel aCtivity of aquaporins by arginine vasopression and atrial natriuretic peptide[J].Biochem Biophys Res Commun,1997,238(7):392-339.
[13]HASEGAWA H,MA T,SKACH W,et al.Molecular cloning of a mercurial-insensitive water channel expressed inselected water-transporting tissues [J].Biol Chem,1994,269(8):5497-5500.
[14]ARIMA H,YAMAMOTO N,SOBUE K,et al.Hyperosmolar mannitol simulates expression of aquaporin 4 and 9 through a p38 mitogen -activated protein kinase-dependent pathway in rat astrocytes[J].J Biol Claem,2003,278(8):44525-44534.
[15]MANLEY GT,FUIMURA M,MA T,et al.Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke[J].Nat Med,2000,6(9):159-163.
[16]AMORINI AM,DUNBAR JG,MARMAROU A.Modulation of aquaporin-4water transport in a model of TBI[J].ACta Neurochir,2003,86(Suppl):261-263.
[17]KE C,POON WS,NG HK,et al.ImpaCt of experimental acute hyponatremia on seer traumatic brain injury in rats:influences on inuries,permeability of blood-brain barrier,ultrastructural features,and aquaporin-4 expression[J].Exp Neurol,2002,178(7):194-206.
[18]KIENING KL,LANDEGHEM FK,SCHREIBER S,et al.Decreased hemispheric Aquaporin-4 is linked to evolving brain edema following controlled cortical impact injury in rats[J].Neurosci Lett,2002,324(8):105-108.
[19]SUN MC,HONEY CR,BERK C,et al.Regulation of aquaporin-4 in a traumatic brain injury model in rats[J].J Neurosurg,2003,98(4):565-569.
[20]SAADOUN S,PAPADOPOULOS MC,KRISHNA S.Water transport becomes uncoupled from k+ siphoning in brain contusion,bacterial meningitis,and brain tumours:immunohistochemical case review[J].J Clin Pathol,2003,56(8):972-975.
[21]BADAUT J,BRUNNET JE,GROLLIMUND L,et al.Aquaporin 1 and aquaprin 4 experession in human brain after subarachnoid hemorrhage and in peritumoral tissue[J].Acta Neurochir,2003,86(Suppl):495-498.
[22]HU H,YAO HT,ZHANG WP,et al.Increased expression of aquaporin-4 in human traumatic brain injury and brain tumors[J].J Zhejiang Univ Sci B,2005,6(5):33-37.
[23]PAPADOPOULOS MC,MANLEY GT,KRISHNA S,et al.Aquaporin-4 facilitates reabsorption of excess luid in vasogenic brain edema[J].FASEB J,2004,18(6):1291-1293.
[24]KE C,POON WS,NG HK,et al.Heterogeneous responses of aquaporin-4 in oedema formation in a replicated severe traumatic brain injury model in rats [J].Neurosci Lett,2001,301(3):21-24.
[25]ZHAO J,MOORE AN,CLIFTON GL,et al.Sulforaphane enhances aquaporin-4expression and decreases cerebral edema following traumatic brain injury[J].J Neurosci Res,2005,82(7):499-506.
[26]GUO Q,SAVEED I,BARONNE LM,et al.Progesterone administration modulates AQP4 expression and edema after traumatic brain injury in mate rats[J].Exp Nenrol,2006,198(2):469-478.
[27]NIERMANN H,AMIRY-MOGHADDAM M,HOLTHOFF K,et al.A novel role of vasopressin in the brain:modulation of activity-dependent water flux in the neocortex[J].J Neurosei,2001,21(9):3045-3051.
[28]VENERO JI,VIZUETE ML.Detailed localization of aquaporin-4 messenger RNA in the CNS:preferential expression in periventricular organs[J].Neuroscience,1999,94(7):239-250.