摘要
背景:复方缬芎提取物中缬草提取物为天然的γ-氨基丁酸受体激动剂,可以松弛脑血管痉挛;川芎提取物通过血脑屏障并增进组织微循环、抑制血小板聚集及5-羟色胺释放。目的:探讨由有效组分配制成的复方缬芎提取物对脑缺血损害的防治效果。设计:完全随机、阴性和阳性对照实验。单位:华中科技大学同济医学院老年医药学研究所。材料:实验于2003-03/2004-05在华中科技大学同济医学院老年医药学研究所完成。动物:①脑组织营养血流灌注实验:选择昆明种小鼠50只,随机分为正常组、溶剂对照(模型)组、川芎嗪50mg/kg组、复方缬芎170mg/kg组及复方缬芎85mg/kg组,每组10只。②动-静脉旁路法血栓形成实验:选择Wistar大鼠50只,随机分为溶剂对照(模型)组、复方丹参5g/kg组、复方缬芎156mg/kg组、复方缬芎94mg/kg组及复方缬芎31.3mg/kg组,每组10只。③大脑中动脉阻断法实验:选择Wistar大鼠60只,随机分为假手术组、溶剂对照(模型)组、川芎嗪10mg/kg组、复方缬芎156mg/kg组、复方缬芎94mg/kg组及复方缬芎31.3mg/kg组,每组10只。④急性广泛性脑缺血实验:选择昆明种小鼠50只,随机分为正常组,溶剂对照(模型)组、川芎嗪10mg/kg组、复方缬芎200mg/kg组及复方缬芎40mg/kg组,每组10只。方法:①脑组织营养血流灌注实验中各组小鼠分别预先腹腔注射复方缬芎(85,170mg/kg)、川芎嗪(50mg/kg)或等容积助溶剂(0.2mL),20min后静脉注射垂体后叶素(2.5u/kg),10min后尾静脉注射同位素99Tcm+亚锡双半胱氨乙酯3.7×1010Bq/L(0.1mL/只)。注射15min后用放射免疫计数仪计取血和大脑组织脉冲强度。②动-静脉旁路法血栓形成实验,术前分别灌胃给复方缬芎156,94,31.3mg/(kg·d),复方丹参注射液[相当生药5g/kg·d)]和生理盐水,连续给药7d,1次/d,停药24h后戊巴比妥钠腹腔麻醉下以体外导流管(内置手术丝线)连接颈总静脉与颈总动脉。体外导流15min后称取血栓质量。③大脑中动脉阻断法实验,水合氯醛腹腔麻醉下,线栓法阻断一侧大脑中动脉,假手术组除不插入线栓外其他手术处理同试验组。各组分别于术前1h、术后3,12h灌胃给复方缬芎(156,94,31.3mg/kg),川芎嗪(10mg/kg)或等容积的助溶剂。手术造模24h后进行神经功能损害行为评分,测定脑质量指数和脑梗死指数。④小鼠急性广泛性脑缺血实验采用胶原蛋白+肾上腺素尾静脉注射造模。各给药组鼠造模注射前30min、注射后1h分别灌胃给予复方缬芎(200,40mg/kg),川芎嗪(10mg/kg)或等容积的助溶剂。观察注射造模后5min内死亡鼠数,15min内偏瘫鼠数,8h后处死测取脑质量指数和紫外分光光度法检测脑组织匀浆的乳酸水平。主要观察指标:①各组小鼠大脑组织/血γ-射线脉冲强度比。②各组大鼠颈总动-静脉体外导流管血栓质量。③各组大鼠神经功能损害行为评分。④各组小鼠脑质量指数。⑤各组大鼠脑梗死指数。⑥各组小鼠死亡率。⑦各组小鼠偏瘫率。⑧各组小鼠脑组织匀浆的乳酸含量。结果:小鼠急性广泛性脑缺血实验中,溶剂对照组造模过程中死亡3只,其余207只实验鼠进入结果分析。①在小鼠脑组织营养血流灌注实验中,脑与血的γ-射线脉冲强度比值:复方缬芎85mg/kg组,复方缬芎170mg/kg高于模型组(0.53±0.09,0.55±0.08,0.45±0.08,t=2.2346,2.7933,P<0.05)。②在大鼠血栓法血小板聚集性实验中,血栓质量:复方缬芎156mg/kg组、复方缬芎94mg/kg组及复方缬芎31.3mg/kg组明显低于模型组([12.66±4.79),(13.31±3.97),(13.49±4.09),(19.21±5.76)g,(t=2.6670,2.5604,2.7649,P<0.05)]。③在大脑中动脉阻塞实验中,复方缬芎31.3mg/kg组、复方缬芎94mg/kg组及复方缬芎156mg/kg组的行为评分明显低于模型组[(5.9±1.9),(6.0±2.0),(5.8±2.2),(8.7±0.9)分],脑梗死指数低于模型组[(16.52±5.78)%,(16.54±3.00)%,(14.18±6.13)%,(24.03±4.85)%,(t=3.1189～4.2118,P<0.01)]。④在小鼠急性广泛性脑缺血实验中,复方缬芎40mg/kg组及复方缬芎200mg/kg组的脑质量指数明显低于模型组[(0.91±0.20)和(0.82±0.24)%,(1.40±0.32)%],脑组织乳酸水平低于模型组[(17.44±6.71),(14.43±2.81),(29.07±7.33)μmol/g,t=3.3885～5.8005,P<0.01]。结论:复方缬芎提取物对垂体后叶素诱导的小鼠脑缺血、对大鼠大脑中动脉栓塞损害有显著的改善作用,对肾上腺素+胶原蛋白混合液或异物诱导的血小板聚集、血栓形成有显著的抑制作用,显示出复方缬芎提取物对脑微循环灌流障碍具有显著的防治作用。
BACKGROUND： In valerian-ligusticum extract （VLE）, valeriana officinalis extract （VOE） is γ aminobutyric acid （GABA） receptor kinetin, which can relax cerebral vascular spasm; ligusticum wallichii Fr. Extraxt （LWE） can pass （hrough blood-brain barrier, enhance microcirculation of tissue and inhibit blood platelet aggregation and 5-Hydroxytryptamine （5-HT） release. OBJECTIVE： To prohe into the effects of VLE prepared with effective components on prevention and treatment of cerebral ischemie injury. DESIGN： Complete randomized, negative and positive control experiment. SETTING： Institute of Senile Medicine and Pharmacology of Tongji Med- ical College of Huazhong University of Science arid Technology. MATERIALS： The experiment was performed in Institute of Senile Medicine Pharmcology of Tonal Medical College of Huazhong University of Science and Teehnology from March 2003 to May 2004. Animals： ① Nutrient blood perfusion in brain tissue： Fifty Kunming mice were employed, which was randomized into normal group, solvent control （model） group, ligustrazine 50 mg/kg group, VLE 170 mg/kg group and VLE 85 mg/kg group, 10 mice in each one. ② Arterial-venous bypass method for thrombosis： Fifty Wistar rats were employed, which was randomized into solvent control （model） group, compound danshen （Radix Salviae Miltiorrhizae） 5 g/kg group, VLE 156 mg/kg group, VLE 94 mg/kg group and VLE 31.3 mg/kg group, 10 rats in each one. ③ Middle cerebral artery occlusion （MCAO） experiment： Sixty Wistar rats were employed, which was randomized into sham-operation group, solvent control （model） group, ligustrazine 10 mg/kg group, VLE 156 mg/kg group, VLE 95 mg/kg group and VLE 31.3 mg/kg, 10 mice in each one.④ Acute extensive cerebral ischemie experiment： Fifty Kunming mice were employed, which was randomized into normal group, solvent contml （model） group, ligustrazine 10 mg/kg group, VLE 200 mg/kg group and VLE 40 mg/kg, 10 mice in each one. METHODS： ① In the experiment of nutrient blood perfusion in brain tissue, in advance, VLE （85, 170 mg/kg）, ligustrazine （50 mg/kg） or solvent enhancer of equal volume （0.2 mL） were injected abdominally in each group. Twenty minutes later, pituitrin （2.5 u/kg） was injected intravenously： and 10 minutes later, isotope ^99Tc^m＋ L, L-EthylCysteinate Dimer and Stannous Chloride （ECD） 3.7×10^10 Bq/ L（0.1 mL/per mouse） was injected in coecygeal nerve. Fifteen minutes later, radio-immunity counter was used to calculate pulsating intensity of blood and cerebral tissue.② In the experiment of arteral-ovenous bypass method for thrombosis, before the operation, gastric perfusion was done with VLE 156, 94, 31.3 mg/（kg.d）, compound danshen injection [equal to 5 g/（kg.d） of raw herb] and physiological saline successively, continuously for 7 days, once per day. After 24 hours of medication pause, with abdominal anesthesia with pentobarbitol sodium, a catheter （with surgical thread inside） was used in vitro to connect common cervical vein and carotid artery. Thrombus mass was sealed 15 minutes after catheterization in vitro. ③ In MCAO experiment, with abdominal anesthesia of chloral hydrate, intraluminal thread approach （ITA） was used to block unilateral MCA. Except that ITA was not used, the other management in sham-operation group was same as experimental groups. Gastric perfusion was done with VLE（156, 94, 31.3 mg/kg）, ligustrazine （10 mg·kg） or solvent enhaneer of equal volume successively 1 hour before operation and 3 hours and 12 hours after operation. 24 hours after modeling, the assessment was done for behavioral neurological damage and brain mass index and cerebra infarction index were observed. ④ In acute extensive cerebral ischemia experiment, the model was prepared by coccygeal injection of collagen ＋ adrenalin （AD）. Respectively, 30 minutes before modeling injection and 1 hour after injection, gastric perfusion was done with VLE （200, 40 mg/kg）, ligustrazine （10 mg/kg） or solvent enhancer of equal volume successively to observe the numbers of dead mice in 5 minutes after modeling and the numbers of hemiplegia mice in 15 minutes; and to determine brain mass index 8 hours later after sacrificed and lactic acid level of brain tissue homogenate with ultraviolet spectrophotometry. MAIN OUTCOME MEASURES： ① Brain tissue/blood γ-pulsating intensity of mice in each group; ② thrombus mass in catheterization of cervical carotid artery and vein in vitro of rats in each group; ③ behavioral neurological assessment （BNA） of rats in each group; ④ brain mass index of mice in each group; ⑤ cerebral infarction index of rats in each group; ⑥ death rate of mice in each group; ⑦ hemiplegia rate of mice in each group; ⑧ lactic acid level in brain tissue homogenate of mice in each group. RESULTS： In the experiment of acute extensive brain ischemia in mice, in solvent control, during modeling, 3 mice were died and the rest 207 mice entered result analysis.① In the experiment of nutrient blood perfusion of brain tissue in mice, the ratios of brain with and blood γ ray pulsating intensity in VLE 85 mg/kg group and VLE 170 mg/kg were higher than model group （0.53±0.09, 0.55±0.08, 0.45±0.08, t=2.234 6, 2.793 3, P 〈 0.05）. （±） In experiment of blood platelet aggregation with thrombosis method in rats, the thrombus masses in VLE 156 mg/kg group, 94 mg/kg group and 31.3 g/kg group were lower remarkably than the model group [（12.66±4.79）, （13.31 ±3.97）, （13.49±4.09）, （19.21±5.76） g, （t=2.667 0, 2.560 4, 2.764 9, P 〈 0.05）]. ③ In MCAO experiment, the BNA in VLE 31.3 mg/kg group, 94 mg/kg group and 156 mg/kg group was lower remarkably than model group successively [（5.9 ±1.9）, （6.0±2.0）, （5.8 ±2.2）, （8.7±0.9） score], and cerebral infarction index was lower than model group [（16.52±5.78）%,（16.54±3.00）%, （14.18±6,13）% , （24.03±4.85）%, （t=3.118 9- 4.211 8, P 〈 0.01）].④ In the experiment of acute extensive cerebral isehemia in mice, brain mass indexes of VLE 40 mg/kg and 200 mg/kg groups were lower remarkably than model group [（0.91±0.20） and （0.82±0.24）%, （1.40±0.32）%], and lactic acid in brain tissue was lower than model group [（17.44±6.71）,（14,43±2.81）, （29,07±7.33） μmol/g （t=3.388 5- 5.800 5, P〈 0.01）]. CONCLUSION： Valerian-liqusticum extract improves significantly cerebral ischemia in mice induced lay pituitrin and the damage by medium cerebral artery embolism in rats. and it inhibits significantly blood platelet aggregation and thrombosis induced by AD＋ collagen mixture or foreign objects. It is suggested that valerian-ligustrazine extract prevents and treats significantly the perfusion disturbance of cerebral microcireulation.
出处
《中国临床康复》
CSCD
北大核心
2005年第33期171-174,共4页
Chinese Journal of Clinical Rehabilitation
