活血利水法治疗创伤性脑水肿的理论与实验研究
|
摘要
1 目 的 本研究根据祖国医学血水相关理论,结合现代医学知识和实验方法,对创伤性脑水肿主要病理生理改变进行研究,探讨创伤性脑水肿发病机理和证治规律及治疗的新途径。利用创伤性脑水肿动物模型,以颅脑损伤后脑微循环改变为切入点,从微血管结构和功能的改变,探讨颅脑损伤后脑微循环紊乱的特点及其对脑血流灌注、血脑屏障通透性及脑水肿等的影响,进一步探讨活血利水法对颅脑损伤后脑微循环的改善及脑水肿的防治作用,为活血利水法治疗创伤性脑水肿提供理论依据及实验基础。通过脑损伤后脑微血管基底膜基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)及星形胶质细胞足突水通道蛋白 4(aquaporin,AQP4)的动态变化探讨脑微血管损伤的机制及活血利水中药脑疏宁对微血管损伤的保护机制。
2 方 法
2.1 按 Feeney 自由落体撞击法建立大鼠创伤性脑损伤(traumatic brain injury,TBI)模型,设假手术组、创伤组、脑疏宁组、甘露醇组。术前 3d 开始连续给予脑疏宁水煎液灌胃,第 4d 给药后 1h 造模;甘露醇组给予 20%甘露醇 1g/kg 股静脉注射。术后 7d 连续动态观察体重及神经功能评分的变化。
2.2 用干/湿重法和 Evans 蓝测定法观察大鼠 TBI 后 6h、24h、3d 及 5d 脑组织含水量和血脑屏障通透性的变化。
2.3 采用血管组化染色及图像分析方法,对大鼠TBI后不同时相的脑微血管分布密度进行动态观察和定量分析,并结合HE染色、透射电镜及MSB染色等技术对脑损伤后脑微血管形态学改变及凝血功能等进行研究。
2.4 采用免疫组化方法观察大鼠 TBI 后不同时相脑组织 MMP-2、MMP-9 及 AQP4的表达。
2.5 运用磁共振成像(magnetic resonance imaging,MRI)技术连续动态观察脑挫伤区信号与体积变化。
3 结 果
3.1 创伤性脑损伤后大鼠出现明显行走功能障碍伴体重减轻。伤后 7d 创伤组大鼠完成木条行走作业的时间为 16.46±3.27s,体重较术前减轻 17.75±5.09g;脑疏宁组行走时间(10.24±1.38s)缩短,与创伤组比较有显著性差异(P<0.01)。体重变化(11.75±4.27g)较小,与创伤组比较有显著性差异(P<0.05);甘露醇组伤后 7d 大鼠完成木条行走作业的时间及体重变化与创伤组比较无显著性差异(P>0.05)。
- 2 - 活血利水法治疗创伤性脑水肿的理论与实验研究
3.2 创伤性脑损伤后 6h 伤侧脑组织含水量(78.76±0.49%)及 EB 含量(10.41±
1.50μg/g)较假手术组(77.20±0.45%,3.27±1.05μg/g)增高,24h 达高峰,分别为
79.63±0.45%和 11.76±2.22μg/g,伤后 5d 未恢复正常。脑疏宁组伤后 6h、24h、3d、
5d 脑组织含水量分别为 78.14±0.37%、78.93±0.47%、78.23±0.63%、77.70±0.33%,
与创伤组同一时间点相比,脑组织含水量降低,有显著性差异(P<0.05);甘露醇组脑组
织含水量分别为 78.01±0.82%、78.80±0.57%、78.26±0.82%、77.81±0.48%,6h、24h、
3d 时间点较创伤组降低,有显著性差异(P<0.05),伤后 5d 两组比较无显著性差异
(P>0.05)。脑疏宁组与甘露醇组同一时间点脑组织含水量比较无显著性差异(P>0.05)。
脑疏宁组脑组织 EB 含量分别为 8.50±1.33μg/g、7.97±1.08μg/g、6.57±1.93μg/g、5.31
±1.19μg/g,与创伤组同一时间点相比,EB 含量明显降低,有显著性差异(P<0.01);
甘露醇组伤后 24h、3d、5d 脑组织 EB 含量与创伤组比较无显著性差异(P>0.05)。
3.3 应用 DAB 组织化学染色,大鼠脑微血管呈棕黄至棕黑色。颅脑损伤后 24h,
脑微血管面积密度(0.021±0.0032%)下降至假手术组(0.051±0.0022%)的 41%,血
管染色浅淡,部分血管有扭曲、断裂现象,血管粗细不均匀;脑疏宁组伤后 24h 开始微
血管面积密度(0.027±0.0056%)明显增加,与创伤组比较有显著性差异(P<0.05)。伤
后 24h 大鼠血浆纤维蛋白原(Fib)及 D-二聚体(D-dimer)含量(3.05±0.71g/L,1.04
±0.32μg/ml)较假手术组增高(2.34±0.23g/L,0.31±0.084μg/ml),提示机体处于高
凝状态并继发纤溶亢进;脑疏宁组 Fib 及 D-dimer 均显著降低(1.98±0.39 g/L,0.52±
0.30μg/ml),与创伤组比较有显著性差异(P<0.05)。
3.4 正常大鼠大脑半球 MMP-2 有少量表达,主要见于脑微血管周围;创伤组大鼠
伤后 6h 伤灶周围 MMP-2 表达开始增高,至伤后 24h,其表达继续增加,5d 时达到高峰;
脑疏宁组与创伤组比较,伤后 24h 开始 MMP-2 表达降低,有显著性差异(P<0.01)。正
常大鼠脑组织未发现 MMP-9 免疫阳性;创伤组大鼠于术后 6h 伤灶周围可见棕黄色
MMP-9 阳性染色细胞,24h 达到高峰,5d 时仍维持较高水平;脑疏宁治疗组与创伤组同
一时间点相比,MMP-9 表达下降,有显著性差异(P<0.01)。
3.5 AQP4 在各组大鼠脑组织中均有表达,阳性表达主要见于脑室和导水管系统的
室管膜、软脑膜及血管周围的星形胶质细胞足突膜上,假手术组呈低表达;TBI 后 6h 伤
灶周围水肿区星形胶质细胞足突 AQP4 表达开始增高,24h 表达继续增加,3d 后逐渐降
低;脑疏宁组大鼠各时间点伤灶周围 AQP4 的表达与创伤组相比明显减低,具有显著性
差异(P<0.05)。
3.6 T2WI 检测显示创伤组大鼠 1d、3d、5d 时间点,右脑额顶叶出现高信号区。伤
后5d
1 Objective
According to the theory of TCM about the relation of blood and water, the knowledge of mordern medicine and mordern experimental method, the present study tried to investigate the pathological and physiological changes of traumatic brain edema in order to identify the pathogenesis, diagnosis and treatment of brain edema. In order to provide theoretical and experimental evidence for promoting blood flow and alleviating water retention to treat brain edema, TBI animal model was used to study the feature of microcirculation disturbance after TBI and its impact on cerebral blood flow, permeability of blood-brain barrier and brain edema. We also tried to investigate the protective effects of promoting blood flow and alleviating water retention on microcirculation and brain edema after TBI. In addition, through observing the change of MMP-2 and MMP-9 on basement membrane and AQP4 on foot process of astrocyte, the mechanism of microvessel injury and the protective mechanism of chinese medicine Naoshuning to microvessel were investigated.
2 Method
2.1 The model of TBI was established by bumpiness of free falling body according to Feeney’s, all experimental rats were devided into pseudoperarion group, TBI group, Naoshuning group and mannitol group. In Naoshuning group, we successively poured into rats’ stomache with Naoshuning for 3 days before operation. 1 hour after the last administration on the 4th day, TBI models were induced. Rats’ weight and neurological function score were detected each day after opertion within one week. Rats in mannitol group were intravenous injected 20% mannitol after operation.
2.2 We observed the variance of permeability of BBB by the method of Evans blue fluorometry. Water content in brain tissue were also evaluated.
2.3 We observed and analyzed microvessel area density by the method of blood vessel histochemical stain and image analysis. We observed the structural and functional change of microvessels with light and eletron microscope. We also detected fibrinogen and D-dimer in order to observe the change of coagulation-fibrinolusis of the blood system.
2.4 Immunohistochemistry was used to observe the brain protein expression of MMP-2,MMP-9 and AQP4 in rats with TBI.
2.5 MRI was successivly used to observe the the change of signal intensity and volume of abnormal high signal region on T2WI after TBI.
3 Result
3.1 After TBI, the beam-walking ability of rats was seriously damaged and the weight of rats was lost obviously. 7 days after TBI, the beam-walking time of rats in TBI group was 16.46±3.27s, the rats’ weight was lost 17.75±5.09g. The beam-walking time of rats in Naoshuning group (10.24±1.38s) significantly decreased compared with TBI group (P<0.01); The change of rats’ weight (11.75±4.27g) was less compared with TBI group (P<0.05). Compared with TBI group, the beam-walking time of rats and the loss of rats’ weight in mannitol group had no significantly difference (P>0.05). 3.2 6 hours after operation, water content of brain tissue (78.76±0.49%) and EB content (10.41±1.50μg/g) in the contusion side significantly increased in TBI group compared with pseudoperarion group (77.20±0.45%, 3.27±1.05μg/g) (P<0.01). They reached summit (79.63±0.45%, 11.76±2.22μg/g) 24 hours after operation and did not come back normal level 5 days after operation. 6h, 24h, 3d, 5d after operation, water content of brain tissue was respectively 78.14±0.37%, 78.93±0.47%, 78.23±0.63%, 77.70±0.33% in Naoshuning group. There was significant difference between Naoshuning group and TBI group (P<0.05 ). In mannitol group, water content of brain tissue was respectively 78.01±0.82%, 78.80±0.57%, 78.26±0.82%, 77.81±0.48%. There was significant difference between mannitol group and TBI group 6h, 24h and 3d after operation (P<0.05). There was no significant difference between Naoshuning group and mannitol group (P>0.05); In Naoshuning group, EB content in contusion brain tissue was respectively 8.50±1.33μg/g, 7.97±1.08μg/g, 6.57±1.93μg/g, 5.31±1.19μg/g. There was signi
2 方 法
2.1 按 Feeney 自由落体撞击法建立大鼠创伤性脑损伤(traumatic brain injury,TBI)模型,设假手术组、创伤组、脑疏宁组、甘露醇组。术前 3d 开始连续给予脑疏宁水煎液灌胃,第 4d 给药后 1h 造模;甘露醇组给予 20%甘露醇 1g/kg 股静脉注射。术后 7d 连续动态观察体重及神经功能评分的变化。
2.2 用干/湿重法和 Evans 蓝测定法观察大鼠 TBI 后 6h、24h、3d 及 5d 脑组织含水量和血脑屏障通透性的变化。
2.3 采用血管组化染色及图像分析方法,对大鼠TBI后不同时相的脑微血管分布密度进行动态观察和定量分析,并结合HE染色、透射电镜及MSB染色等技术对脑损伤后脑微血管形态学改变及凝血功能等进行研究。
2.4 采用免疫组化方法观察大鼠 TBI 后不同时相脑组织 MMP-2、MMP-9 及 AQP4的表达。
2.5 运用磁共振成像(magnetic resonance imaging,MRI)技术连续动态观察脑挫伤区信号与体积变化。
3 结 果
3.1 创伤性脑损伤后大鼠出现明显行走功能障碍伴体重减轻。伤后 7d 创伤组大鼠完成木条行走作业的时间为 16.46±3.27s,体重较术前减轻 17.75±5.09g;脑疏宁组行走时间(10.24±1.38s)缩短,与创伤组比较有显著性差异(P<0.01)。体重变化(11.75±4.27g)较小,与创伤组比较有显著性差异(P<0.05);甘露醇组伤后 7d 大鼠完成木条行走作业的时间及体重变化与创伤组比较无显著性差异(P>0.05)。
- 2 - 活血利水法治疗创伤性脑水肿的理论与实验研究
3.2 创伤性脑损伤后 6h 伤侧脑组织含水量(78.76±0.49%)及 EB 含量(10.41±
1.50μg/g)较假手术组(77.20±0.45%,3.27±1.05μg/g)增高,24h 达高峰,分别为
79.63±0.45%和 11.76±2.22μg/g,伤后 5d 未恢复正常。脑疏宁组伤后 6h、24h、3d、
5d 脑组织含水量分别为 78.14±0.37%、78.93±0.47%、78.23±0.63%、77.70±0.33%,
与创伤组同一时间点相比,脑组织含水量降低,有显著性差异(P<0.05);甘露醇组脑组
织含水量分别为 78.01±0.82%、78.80±0.57%、78.26±0.82%、77.81±0.48%,6h、24h、
3d 时间点较创伤组降低,有显著性差异(P<0.05),伤后 5d 两组比较无显著性差异
(P>0.05)。脑疏宁组与甘露醇组同一时间点脑组织含水量比较无显著性差异(P>0.05)。
脑疏宁组脑组织 EB 含量分别为 8.50±1.33μg/g、7.97±1.08μg/g、6.57±1.93μg/g、5.31
±1.19μg/g,与创伤组同一时间点相比,EB 含量明显降低,有显著性差异(P<0.01);
甘露醇组伤后 24h、3d、5d 脑组织 EB 含量与创伤组比较无显著性差异(P>0.05)。
3.3 应用 DAB 组织化学染色,大鼠脑微血管呈棕黄至棕黑色。颅脑损伤后 24h,
脑微血管面积密度(0.021±0.0032%)下降至假手术组(0.051±0.0022%)的 41%,血
管染色浅淡,部分血管有扭曲、断裂现象,血管粗细不均匀;脑疏宁组伤后 24h 开始微
血管面积密度(0.027±0.0056%)明显增加,与创伤组比较有显著性差异(P<0.05)。伤
后 24h 大鼠血浆纤维蛋白原(Fib)及 D-二聚体(D-dimer)含量(3.05±0.71g/L,1.04
±0.32μg/ml)较假手术组增高(2.34±0.23g/L,0.31±0.084μg/ml),提示机体处于高
凝状态并继发纤溶亢进;脑疏宁组 Fib 及 D-dimer 均显著降低(1.98±0.39 g/L,0.52±
0.30μg/ml),与创伤组比较有显著性差异(P<0.05)。
3.4 正常大鼠大脑半球 MMP-2 有少量表达,主要见于脑微血管周围;创伤组大鼠
伤后 6h 伤灶周围 MMP-2 表达开始增高,至伤后 24h,其表达继续增加,5d 时达到高峰;
脑疏宁组与创伤组比较,伤后 24h 开始 MMP-2 表达降低,有显著性差异(P<0.01)。正
常大鼠脑组织未发现 MMP-9 免疫阳性;创伤组大鼠于术后 6h 伤灶周围可见棕黄色
MMP-9 阳性染色细胞,24h 达到高峰,5d 时仍维持较高水平;脑疏宁治疗组与创伤组同
一时间点相比,MMP-9 表达下降,有显著性差异(P<0.01)。
3.5 AQP4 在各组大鼠脑组织中均有表达,阳性表达主要见于脑室和导水管系统的
室管膜、软脑膜及血管周围的星形胶质细胞足突膜上,假手术组呈低表达;TBI 后 6h 伤
灶周围水肿区星形胶质细胞足突 AQP4 表达开始增高,24h 表达继续增加,3d 后逐渐降
低;脑疏宁组大鼠各时间点伤灶周围 AQP4 的表达与创伤组相比明显减低,具有显著性
差异(P<0.05)。
3.6 T2WI 检测显示创伤组大鼠 1d、3d、5d 时间点,右脑额顶叶出现高信号区。伤
后5d
1 Objective
According to the theory of TCM about the relation of blood and water, the knowledge of mordern medicine and mordern experimental method, the present study tried to investigate the pathological and physiological changes of traumatic brain edema in order to identify the pathogenesis, diagnosis and treatment of brain edema. In order to provide theoretical and experimental evidence for promoting blood flow and alleviating water retention to treat brain edema, TBI animal model was used to study the feature of microcirculation disturbance after TBI and its impact on cerebral blood flow, permeability of blood-brain barrier and brain edema. We also tried to investigate the protective effects of promoting blood flow and alleviating water retention on microcirculation and brain edema after TBI. In addition, through observing the change of MMP-2 and MMP-9 on basement membrane and AQP4 on foot process of astrocyte, the mechanism of microvessel injury and the protective mechanism of chinese medicine Naoshuning to microvessel were investigated.
2 Method
2.1 The model of TBI was established by bumpiness of free falling body according to Feeney’s, all experimental rats were devided into pseudoperarion group, TBI group, Naoshuning group and mannitol group. In Naoshuning group, we successively poured into rats’ stomache with Naoshuning for 3 days before operation. 1 hour after the last administration on the 4th day, TBI models were induced. Rats’ weight and neurological function score were detected each day after opertion within one week. Rats in mannitol group were intravenous injected 20% mannitol after operation.
2.2 We observed the variance of permeability of BBB by the method of Evans blue fluorometry. Water content in brain tissue were also evaluated.
2.3 We observed and analyzed microvessel area density by the method of blood vessel histochemical stain and image analysis. We observed the structural and functional change of microvessels with light and eletron microscope. We also detected fibrinogen and D-dimer in order to observe the change of coagulation-fibrinolusis of the blood system.
2.4 Immunohistochemistry was used to observe the brain protein expression of MMP-2,MMP-9 and AQP4 in rats with TBI.
2.5 MRI was successivly used to observe the the change of signal intensity and volume of abnormal high signal region on T2WI after TBI.
3 Result
3.1 After TBI, the beam-walking ability of rats was seriously damaged and the weight of rats was lost obviously. 7 days after TBI, the beam-walking time of rats in TBI group was 16.46±3.27s, the rats’ weight was lost 17.75±5.09g. The beam-walking time of rats in Naoshuning group (10.24±1.38s) significantly decreased compared with TBI group (P<0.01); The change of rats’ weight (11.75±4.27g) was less compared with TBI group (P<0.05). Compared with TBI group, the beam-walking time of rats and the loss of rats’ weight in mannitol group had no significantly difference (P>0.05). 3.2 6 hours after operation, water content of brain tissue (78.76±0.49%) and EB content (10.41±1.50μg/g) in the contusion side significantly increased in TBI group compared with pseudoperarion group (77.20±0.45%, 3.27±1.05μg/g) (P<0.01). They reached summit (79.63±0.45%, 11.76±2.22μg/g) 24 hours after operation and did not come back normal level 5 days after operation. 6h, 24h, 3d, 5d after operation, water content of brain tissue was respectively 78.14±0.37%, 78.93±0.47%, 78.23±0.63%, 77.70±0.33% in Naoshuning group. There was significant difference between Naoshuning group and TBI group (P<0.05 ). In mannitol group, water content of brain tissue was respectively 78.01±0.82%, 78.80±0.57%, 78.26±0.82%, 77.81±0.48%. There was significant difference between mannitol group and TBI group 6h, 24h and 3d after operation (P<0.05). There was no significant difference between Naoshuning group and mannitol group (P>0.05); In Naoshuning group, EB content in contusion brain tissue was respectively 8.50±1.33μg/g, 7.97±1.08μg/g, 6.57±1.93μg/g, 5.31±1.19μg/g. There was signi
引文
[1] 张学文.中风病防治研究.陕西科学技术出版社,1998,157.
[2] 黄融琪,张方东.中风急性期脑水肿救治体会.福建中医药,1994,25(3):27.
[3] 孙岩.出血性中风急性期中医治法研究.浙江中医学院学报,1999,23 (2):14.
[4] 杨爱学.醒脑化痰通腑饮治疗中风急性期脑水肿的临床研究.河南中医药学刊, 1994,9(3):39.
[5] 刘建忠.中医药治疗中风脑水肿初探.福建中医药,1997,28 (6):17.
[6] 吕冰,陈红旗,赵勇刚等.丹参治疗重型颅脑损伤的临床研究.医师进修杂志,1999,22(1):19.
[7] 游恒星,邱建东,杨维,等.超早期应用丹参注射液治疗颅脑损伤的临床研究.中国临床神经外科杂志,2003,8( 2 ):117-119.
[8] 周仲瑛, 周 珉, 金妙文,等.中国中西医结合急救杂志,2002, 9(5): 276-278.
[9] 吴宏生,张惠莲,韩玉晶.息风化瘀汤治疗急性脑出 50 例观察.实用中医药杂志,2002,18(2)∶5.
[10] 李军.张学文教授论颅脑水瘀证治疗.中国中医急症,1993,2(5):2-9.
[11] 曾祥发,陆晖,谢林等.参麦注射液对大鼠伤寒内毒素的作用观察. 广西中医药,1989,12(1):44.
[12] 史代双.益气温阳利水汤治疗老年人脑出血脑水肿 40 例报告.实用中西医结合杂志,1998,11(8):203.
[13] 崔向宁.活血利水通脉饮治疗急性中风的研究.山东中医药大学学报,1997,21(1):33-38.
[14] 张永全,刘泰,陆晖等.健神利水1号对脑出血急性期脑水肿的临床疗效.广西中医药,2002,25(5):8-10.
[15] 张永全,刘泰,陆晖等.健神利水1号对脑出血急性期血液流变性及超氧化物歧化酶的影响.中国中西医结合急救杂志,2002,9(5):273-275.
[16] 高树良,王晋朝.利水逐瘀法治疗高血压脑出血颅内压增高临床研究.河北中医,2002,24(4):243-245.
[17] 张昌荣,黑巧红.中西医结合治疗脑水肿 100 例. 陕西中医,1994,15(3):108.
[18] 陈绍宏,张晓云,刘永家.逐瘀化痰口服液治疗急性脑出血的临床与实验研究.中国中医急症,1995,4(2):58.
[19] 李澎涛,王永炎,黄启福.“毒损脑络”病机假说的形成及其理论与实践意义.北京中医药大学学报,2001,24(1):1-6.
[20] 王吕俊,丁寿禅.试论下法治疗急性脑血管病的作用机制和运用原则.山西中医,1992,8(2):6.
[21] 曾大方.略谈腑气不通在中风病中的地位及临床意义.吉林中医药,1984 (1): 14.
[22] 赵江民.降逆复生液治疗急性脑出血30例前后CT对照研究.云南中医药杂志,1999,20(2):12.
[23] 鞠作泉,李忠刚,王风霞.化痰通腑活血法治疗重症出血性中风 62 例.中国中医药息杂志,2005,12(2):78-79.
[24] 朱连科,魏文阁.中西医结合治疗急性脑血管病脑水肿 43 例.现代中西医结合杂志,2000,9(4):331.
[25] 张建军,朱镇宇,张俊等.醒脑静注射液在治疗重症颅脑损伤中抗高热和促醒作 用的疗效观察.中国中西医结合急救杂志,1999,6(1):57.
[26] 王建军,高长庆,陈文武.β-七叶皂苷钠治疗急性脑出血的临床观察. 河南大学学报(医学版),2004,23(4):47-48.
[27] 陈中栋,沈和平.七叶皂甙钠与甘露醇联合治疗脑出血临床分析.浙江临床医学,2002,4(9):668.
[28] 陈立华,刘运生,马建荣等.β-七叶皂甙钠和甘露醇及地塞米松治疗放射性脑水肿的比较.中国神经精神疾病杂志,1999,25(3):150-151.
[29] 郑红艳,陈莉.三七总皂甙治疗急性重型颅脑损伤的疗效观察及护理.中华实用中西医杂志,2004,4(17):1804-1805.
[30] 朱玲,杨术真,杨喜民等. 川芎嗪注射液对急性颅脑损伤患者血液流变性的影响.中国中西医结合急救杂志,1996,(1):28-30.
[31] 陈礼刚,曾凡俊,魏东等.刺五加注射液对重型颅脑损伤患者的疗效观察.第三军医大学学报,2001,23(11):1371-1372.
[32] 乔普,杨养贤,陈明霞等. 活血效灵丹对脑出血兔血脑屏障及脑组织超微结构的影响.西安医科大学学报,2001,23(3):300.
[33] 刘建辉,冀凤云,王婷等. 三七总皂苷对实验性脑缺血脑血流及血脑屏障的影响作用.河北医药,2002,24(4):249.
[34] 李克玲,黄启福,浆玉凤等. 清开灵注射液治疗家兔脑血肿的实验研究. 中国中西医结合杂志,1997,17(2):91.
[35] 杨波,关方霞,刘书君等. 复方丹参注射液对兔脑外伤后经颅多普勒和脑水肿得影响.中国医学影像技术,2000,16(6):502.
[36] 杨喜民,李拴德,王晓峰等. 川芎嗪对急性颅脑损伤大鼠治疗作用的实验研究. 中国中西医结合急救杂志,2001,8(6):360-361.
[37] 庞鹤,王晓梅,王硕仁等. 醒脑健神丹合清开灵注射液治疗急性脑出血的实验研究. 中国中医急症,1995,4(2):69.
[38] 黄世敬,黄启福,孙塑伦等.救脑宁注射液对实验性脑出血大鼠脑组织内皮素含量及一氧化氮合酶的影响.中国中医基础医学杂志,2000,6(8):16.
[39] 童建国,王先荣,朱刚等. 活血化瘀中药对脑挫伤的治疗作用及其机制的实验研究. 第三军医大学学报,2001,23(7):821.
[40] 韩金安,匡永勤,周虎田等.三七总皂甙对家兔脑损伤后脑组织中丙二醛含量变化 的影响.中国病理生理杂志,2000,16(3):269-271.
[41] 张秋菊,王永成,程惠娟等.通腑健脑液对大鼠颅脑损伤治疗作用的实验研究.中国中医急症,2004,13(5):309-310.
[42] 杨于嘉等.黄芩甙对百日咳菌液致大鼠脑水肿的保护作用.中华医学杂志,1998,78(8):4-7.
[43] 谭峰,欧正武,李安国等.补阳还五汤对脑水肿得钙拮抗剂作用.中国危重病急救医学,1995,7(3):136.
[44] 陶永光,岳少杰,俞燕等.清开灵对大鼠谷氨酸神经毒性脑水肿时突触体游离钙的影响.中国当代儿科杂志,2000,2(5):326.
[45] 韩金安,匡永勤,周虎田等.三七总皂甙对颅脑损伤后Ca2+、CaM含量影响的实验研究.中华创伤杂志,1999,15(4)):278- 280.
[46] 赖真,王沙燕,耿小茵等.补阳还五汤和黄芪对沙土鼠脑缺血再灌注后脑组织兴奋性氨基酸的影响.北京中医药大学学报,2002,25(3):33-35.
[47] 邱根全,石秦东,蔡云等.益气活血开窍复方对缺血再灌注脑组织兴奋性氨基酸及超微结构的影响.西安交通大学学报( 医学版),2002,23(5):489-492.
[48] 危静,肖国民.β-七叶皂甙治疗创伤性脑水肿分子作用机制的实验研究.现代中西医结合杂志,2005,14(2):160-162.
[49] 林 海,刘 煜,李柱一等.局灶性脑缺血再灌注后白细胞介素-6、肿瘤坏死因子的表达及川芎嗪对其表达的影响.中国临床康复,2002,6(7):968-969.
[50] 刘勇,许彦钢,林奇. 川芎嗪对脑血管内皮细胞ICAM-1表达的影响.西安医科大学学报,1998,19(2):148-150.
[51] 张兆辉,余绍祖.川芎嗪对局灶性脑缺血多形核白细胞功能的影响.浙江中西医结合杂志,2002年,12(3):154-156.
[52] 王建国,陈群,曾因明. 灯盏花素注射液对沙土鼠脑缺血再灌注后能量代谢及脑水肿的影响.中国中西医结合急救杂志,2004,11(1):25-27.
[53] 张红,朱志安,陈鑫.灯盏花素对大鼠颅脑损伤脑组织线粒体ATP酶活性的影响.中国临床神经外科杂志,2002,7(3):170-171.
[54]Rosenberg GA,Seremin O,Estrada E. Arginine vasopressin V1-antagonist and atrial natriuretic peptide reduce hemorrhage brain edema in rats.Stroke ,1992,23(12):1767-1773.
[55 ] 陈旭,郑惠民,由振东等.七叶皂苷钠对大鼠脑出血后脑水肿及脑内精氨酸加压素含量的影响.第二军医大学学报,2001,22(12):1142-1144.
[56] 沈强,刘亚敏,徐秋英.芳香开窍对缺血再灌注大鼠水通道蛋白-4mRNA 表达的影响.中医药学刊,2003,21(6):871-87.
[57] 高颖,蔡定芳.大脑中动脉阻塞大鼠 MMP 9 基因表达及补阳还五汤对其影响.中成药,2003,25(7):560-563.
[58] Lee KR ,et al.Intracerebral infusion of thrombin as a cause of braine dema. J Neurosurg,1995,83:1045-1050.
[59] Lee KR ,et al.The role of the coagulation cascade in brain edema formation after intracerebral hemorrhage.Acta Neurochir,1996,138(4):396-400.
[60] 吴文斌,胡长林,郭富强等.水蛭提取液对实验性脑内血肿吸收的病理学及动物行为的影响.中国临床康复,2004,8(1):121-123.
[61] 董少龙,黄立武,张茂林。 水蛭提取液对实验性家兔颅内血肿的影响. 广西中医药,2000,23(2):49-50.
[2] 黄融琪,张方东.中风急性期脑水肿救治体会.福建中医药,1994,25(3):27.
[3] 孙岩.出血性中风急性期中医治法研究.浙江中医学院学报,1999,23 (2):14.
[4] 杨爱学.醒脑化痰通腑饮治疗中风急性期脑水肿的临床研究.河南中医药学刊, 1994,9(3):39.
[5] 刘建忠.中医药治疗中风脑水肿初探.福建中医药,1997,28 (6):17.
[6] 吕冰,陈红旗,赵勇刚等.丹参治疗重型颅脑损伤的临床研究.医师进修杂志,1999,22(1):19.
[7] 游恒星,邱建东,杨维,等.超早期应用丹参注射液治疗颅脑损伤的临床研究.中国临床神经外科杂志,2003,8( 2 ):117-119.
[8] 周仲瑛, 周 珉, 金妙文,等.中国中西医结合急救杂志,2002, 9(5): 276-278.
[9] 吴宏生,张惠莲,韩玉晶.息风化瘀汤治疗急性脑出 50 例观察.实用中医药杂志,2002,18(2)∶5.
[10] 李军.张学文教授论颅脑水瘀证治疗.中国中医急症,1993,2(5):2-9.
[11] 曾祥发,陆晖,谢林等.参麦注射液对大鼠伤寒内毒素的作用观察. 广西中医药,1989,12(1):44.
[12] 史代双.益气温阳利水汤治疗老年人脑出血脑水肿 40 例报告.实用中西医结合杂志,1998,11(8):203.
[13] 崔向宁.活血利水通脉饮治疗急性中风的研究.山东中医药大学学报,1997,21(1):33-38.
[14] 张永全,刘泰,陆晖等.健神利水1号对脑出血急性期脑水肿的临床疗效.广西中医药,2002,25(5):8-10.
[15] 张永全,刘泰,陆晖等.健神利水1号对脑出血急性期血液流变性及超氧化物歧化酶的影响.中国中西医结合急救杂志,2002,9(5):273-275.
[16] 高树良,王晋朝.利水逐瘀法治疗高血压脑出血颅内压增高临床研究.河北中医,2002,24(4):243-245.
[17] 张昌荣,黑巧红.中西医结合治疗脑水肿 100 例. 陕西中医,1994,15(3):108.
[18] 陈绍宏,张晓云,刘永家.逐瘀化痰口服液治疗急性脑出血的临床与实验研究.中国中医急症,1995,4(2):58.
[19] 李澎涛,王永炎,黄启福.“毒损脑络”病机假说的形成及其理论与实践意义.北京中医药大学学报,2001,24(1):1-6.
[20] 王吕俊,丁寿禅.试论下法治疗急性脑血管病的作用机制和运用原则.山西中医,1992,8(2):6.
[21] 曾大方.略谈腑气不通在中风病中的地位及临床意义.吉林中医药,1984 (1): 14.
[22] 赵江民.降逆复生液治疗急性脑出血30例前后CT对照研究.云南中医药杂志,1999,20(2):12.
[23] 鞠作泉,李忠刚,王风霞.化痰通腑活血法治疗重症出血性中风 62 例.中国中医药息杂志,2005,12(2):78-79.
[24] 朱连科,魏文阁.中西医结合治疗急性脑血管病脑水肿 43 例.现代中西医结合杂志,2000,9(4):331.
[25] 张建军,朱镇宇,张俊等.醒脑静注射液在治疗重症颅脑损伤中抗高热和促醒作 用的疗效观察.中国中西医结合急救杂志,1999,6(1):57.
[26] 王建军,高长庆,陈文武.β-七叶皂苷钠治疗急性脑出血的临床观察. 河南大学学报(医学版),2004,23(4):47-48.
[27] 陈中栋,沈和平.七叶皂甙钠与甘露醇联合治疗脑出血临床分析.浙江临床医学,2002,4(9):668.
[28] 陈立华,刘运生,马建荣等.β-七叶皂甙钠和甘露醇及地塞米松治疗放射性脑水肿的比较.中国神经精神疾病杂志,1999,25(3):150-151.
[29] 郑红艳,陈莉.三七总皂甙治疗急性重型颅脑损伤的疗效观察及护理.中华实用中西医杂志,2004,4(17):1804-1805.
[30] 朱玲,杨术真,杨喜民等. 川芎嗪注射液对急性颅脑损伤患者血液流变性的影响.中国中西医结合急救杂志,1996,(1):28-30.
[31] 陈礼刚,曾凡俊,魏东等.刺五加注射液对重型颅脑损伤患者的疗效观察.第三军医大学学报,2001,23(11):1371-1372.
[32] 乔普,杨养贤,陈明霞等. 活血效灵丹对脑出血兔血脑屏障及脑组织超微结构的影响.西安医科大学学报,2001,23(3):300.
[33] 刘建辉,冀凤云,王婷等. 三七总皂苷对实验性脑缺血脑血流及血脑屏障的影响作用.河北医药,2002,24(4):249.
[34] 李克玲,黄启福,浆玉凤等. 清开灵注射液治疗家兔脑血肿的实验研究. 中国中西医结合杂志,1997,17(2):91.
[35] 杨波,关方霞,刘书君等. 复方丹参注射液对兔脑外伤后经颅多普勒和脑水肿得影响.中国医学影像技术,2000,16(6):502.
[36] 杨喜民,李拴德,王晓峰等. 川芎嗪对急性颅脑损伤大鼠治疗作用的实验研究. 中国中西医结合急救杂志,2001,8(6):360-361.
[37] 庞鹤,王晓梅,王硕仁等. 醒脑健神丹合清开灵注射液治疗急性脑出血的实验研究. 中国中医急症,1995,4(2):69.
[38] 黄世敬,黄启福,孙塑伦等.救脑宁注射液对实验性脑出血大鼠脑组织内皮素含量及一氧化氮合酶的影响.中国中医基础医学杂志,2000,6(8):16.
[39] 童建国,王先荣,朱刚等. 活血化瘀中药对脑挫伤的治疗作用及其机制的实验研究. 第三军医大学学报,2001,23(7):821.
[40] 韩金安,匡永勤,周虎田等.三七总皂甙对家兔脑损伤后脑组织中丙二醛含量变化 的影响.中国病理生理杂志,2000,16(3):269-271.
[41] 张秋菊,王永成,程惠娟等.通腑健脑液对大鼠颅脑损伤治疗作用的实验研究.中国中医急症,2004,13(5):309-310.
[42] 杨于嘉等.黄芩甙对百日咳菌液致大鼠脑水肿的保护作用.中华医学杂志,1998,78(8):4-7.
[43] 谭峰,欧正武,李安国等.补阳还五汤对脑水肿得钙拮抗剂作用.中国危重病急救医学,1995,7(3):136.
[44] 陶永光,岳少杰,俞燕等.清开灵对大鼠谷氨酸神经毒性脑水肿时突触体游离钙的影响.中国当代儿科杂志,2000,2(5):326.
[45] 韩金安,匡永勤,周虎田等.三七总皂甙对颅脑损伤后Ca2+、CaM含量影响的实验研究.中华创伤杂志,1999,15(4)):278- 280.
[46] 赖真,王沙燕,耿小茵等.补阳还五汤和黄芪对沙土鼠脑缺血再灌注后脑组织兴奋性氨基酸的影响.北京中医药大学学报,2002,25(3):33-35.
[47] 邱根全,石秦东,蔡云等.益气活血开窍复方对缺血再灌注脑组织兴奋性氨基酸及超微结构的影响.西安交通大学学报( 医学版),2002,23(5):489-492.
[48] 危静,肖国民.β-七叶皂甙治疗创伤性脑水肿分子作用机制的实验研究.现代中西医结合杂志,2005,14(2):160-162.
[49] 林 海,刘 煜,李柱一等.局灶性脑缺血再灌注后白细胞介素-6、肿瘤坏死因子的表达及川芎嗪对其表达的影响.中国临床康复,2002,6(7):968-969.
[50] 刘勇,许彦钢,林奇. 川芎嗪对脑血管内皮细胞ICAM-1表达的影响.西安医科大学学报,1998,19(2):148-150.
[51] 张兆辉,余绍祖.川芎嗪对局灶性脑缺血多形核白细胞功能的影响.浙江中西医结合杂志,2002年,12(3):154-156.
[52] 王建国,陈群,曾因明. 灯盏花素注射液对沙土鼠脑缺血再灌注后能量代谢及脑水肿的影响.中国中西医结合急救杂志,2004,11(1):25-27.
[53] 张红,朱志安,陈鑫.灯盏花素对大鼠颅脑损伤脑组织线粒体ATP酶活性的影响.中国临床神经外科杂志,2002,7(3):170-171.
[54]Rosenberg GA,Seremin O,Estrada E. Arginine vasopressin V1-antagonist and atrial natriuretic peptide reduce hemorrhage brain edema in rats.Stroke ,1992,23(12):1767-1773.
[55 ] 陈旭,郑惠民,由振东等.七叶皂苷钠对大鼠脑出血后脑水肿及脑内精氨酸加压素含量的影响.第二军医大学学报,2001,22(12):1142-1144.
[56] 沈强,刘亚敏,徐秋英.芳香开窍对缺血再灌注大鼠水通道蛋白-4mRNA 表达的影响.中医药学刊,2003,21(6):871-87.
[57] 高颖,蔡定芳.大脑中动脉阻塞大鼠 MMP 9 基因表达及补阳还五汤对其影响.中成药,2003,25(7):560-563.
[58] Lee KR ,et al.Intracerebral infusion of thrombin as a cause of braine dema. J Neurosurg,1995,83:1045-1050.
[59] Lee KR ,et al.The role of the coagulation cascade in brain edema formation after intracerebral hemorrhage.Acta Neurochir,1996,138(4):396-400.
[60] 吴文斌,胡长林,郭富强等.水蛭提取液对实验性脑内血肿吸收的病理学及动物行为的影响.中国临床康复,2004,8(1):121-123.
[61] 董少龙,黄立武,张茂林。 水蛭提取液对实验性家兔颅内血肿的影响. 广西中医药,2000,23(2):49-50.