【作者】 桂玲；

【导师】 郭莲军；

【作者基本信息】 华中科技大学 ， 药理学， 2006， 硕士

【摘要】 研究背景与目的缺血性脑血管病,亦称脑卒中,是严重危害人类健康的疾病之一。其发病机理主要是,由于各种原因所致的血管狭窄和血液流变学异常导致血液淤积或血栓形成,最终发展为脑梗塞。以中医中药治疗脑卒中、脑中风类疾病已有悠久历史,抗脑血栓片(Kang Naoxueshuan Tablet,KNT),针对缺血性脑血管病,是由红花、当归、水蛭、赤芍、桃仁、川芎、丹参、土鳖虫、羚羊角、牛黄等传统中药组成,经由现代科技方法提取制成的片剂。其复方组成成分经证实,具有抗血小板、抗凝血和抑制血栓形成的作用。该方剂的主要功能主治:活血化瘀,醒脑通络,潜阳熄风,用于因瘀血、肝阳上亢所致的中风先兆,以及脑血栓形成。本研究采用大鼠大脑中动脉栓塞(MCAO)局灶性脑缺血模型,观察抗脑血栓片对缺血性脑血管病的保护作用,并初步探讨其发挥抗脑缺血作用的机理,为该复方制剂的进一步开发应用提供实验依据。方法1.本研究以线栓法制备大鼠大脑中动脉栓塞(middle cerebral artery occlusion, MCAO)局灶性脑缺血模型,分别在缺血后3h,6h和24h评价大鼠的神经功能状态,并于缺血24h时,处死大鼠取脑测脑梗死体积。观察不同剂量抗脑血栓片混悬液对缺血后大鼠神经运动功能症状的改善以及减少脑梗死体积的作用。2.采用线栓法,通过大鼠颈动-静脉旁路,制备大鼠体外血栓,对血栓的干重和湿重进行称量。观察服用不同剂量的抗脑血栓片混悬液对血栓干、湿重的影响。3.经大鼠颈动脉取血抗凝,观察不同剂量抗脑血栓片混悬液对大鼠血小板计数和血小板聚集功能以及对大鼠红细胞压积、血浆粘度和全血粘度的影响。在MCAO模型和体外血栓模型实验中,均设模型组(或空白对照组)、阳性药组、原料药组和药物治疗组。模型组(或空白对照组)动物给予等容积生理盐水;阳性药组给予消栓通(中国药典已收录);原料药组给予抗脑血栓片干浸膏粉;治疗组分为低、中、高剂量三个剂量并配制成相应浓度;各组动物均按等体积不等浓度灌胃给与。结果1.模型组大鼠缺血后出现明显的神经运动功能障碍,神经症状随缺血时间延长而加重,评分分别为:3h,2.80±1.3;6h,3.00±1.33;24h,3.56±1.5。与模型组相比,抗脑血栓片各剂量对神经症状均有明显的改善作用,且作用随剂量的增加而增强:低剂量组评分,3h,1.44±0.53;6h,1.56±0.53;24h,1.67±0.87(P<0.01);中剂量组评分,3h,1.10±0.57;6h,1.30±0.48;24h,1.40±0.69(P<0.01);高剂量组评分,3h,0.90±0.44;6h,1.00±0.71;24h,1.30±0.53(P<0.01)。缺血24h后,与模型组脑梗死体积(27.51±4.71)%相比,抗脑血栓片各剂量组均可减少脑梗死体积百分比:低剂量组,(15.37±7.21)%(P<0.01);中剂量组,(14.60±7.00)%(P<0.01);高剂量组,(11.18±3.35)%(P<0.01)。2.结果显示,抗脑血栓片各剂量组均可明显减轻体外血栓的干、湿重。与模型组血栓干重4.93±2.17mg相比,治疗组血栓干重量分别为3.35±1.44,3.13±1.38mg(P<0.05),2.88±1.02mg(P<0.05)。模型组血栓湿重为24.23±8.21mg,治疗组血栓的湿重分别为,15.56±5.13mg(P<0.05), 12.26±3.55mg(P<0.01),11.08±3.10mg(P<0.01)。3.与对照组比较,抗脑血栓片高剂量给药后,对ADP诱导的血小板聚集有抑制作用,对1、3、5分钟各时间点聚集率和最大聚集率的抑制率分别为(40.24±12.9)%,(43.08±5.44)%,(56.43±7.93)%和(42.13±8.45)%(P<0.01)。而中剂量组只在1min的时间点与对照组比较有显著差异(P<0.05);低剂量组各时间点与对照组比较,均无显著差异。对血小板计数,仅高剂量组会降低血小板计数(P<0.05),中和低剂量组对血小板计数均无明显影响。4.对大鼠红细胞压积和血浆粘度,各治疗组均未表现出明显作用。而在37.5/s, 90/s和300/s切变率下的全血粘度,与对照组比较,中剂量组可使其分别降低(36.03±5.45)%,(41.41±7.71)%和(32.97±6.17)%(P<0.01),高剂量组则可使其分别降低(30.31±11.44)%,(35.51±8.37)%和(19.84±3.80)%(P<0.05)。结论以上药效学实验结果表明,抗脑血栓片可改善大鼠局灶性脑缺血后神经功能状态,减少脑梗死体积,对脑缺血有一定的保护作用。其作用机制可能与其降低全血粘度、抑制血小板聚集,抑血栓形成有关。更多还原

【Abstract】 Background and purpose Ischemia cerebral injury is also named stroke by traditional chinese physician. And the experience in treating this kind disease has been accumulated in the long history of the applicant of the traditional chinese medicine. Complex reasons cause angiostegnosis and the abnormity of blood rheology , whereafter induce the blood siltation and thrombogenesis, in the ultimate the cerebral infarction is formed. That is the main pathomechanism of ischemia cerebral injury. Kang Naoxueshuan Tablet(KNT), which is composed by many chinese crude drug and extracted through advanced technology, can treat presymptom of stroke and cerebral thrombosis.And its complex moity have been proved effective to antipatelt、anticoagulated blood and suppress thrombogenesis. In this thesis ,we applied the focal cerebral ischemia model by Middle cerebral artery occlusion (MCAO) in rat, observed the protective effect and approached the machnism of KNT againt ischmia cerebral injury. Our aim is to get more experimental evidance to support the exploitation and application of the compound preparation.Methods1. Focal cerebral ischemia model in rat was induced by transient occlusion of the middle cerebral artery . After MCAO, evaluated neurological deficit score in 3h,6h and 24h, and sacrificed rats after 24h to measure infarct volum.Observed effect of different dosage suspension of KNT on improving neuromotor function and reducing brain infarct volum in rat.2. Formed the bypass between arteria cervicalis and vena cervicalis, and prepared the vitro-thrombus in rats by thread-embolism method. Weighed the thrombus in dry and wet status respectively. Observed the effect of KNT suspension on thrombus dry and wet weight.3. To get the anticoagulated blood through arteria cervicalis cannula in rats, evaluated effect of different dosage suspension of KNT on blood platelets count and aggregation of platelet, and effect on HCT、PV and WBV.In the model of MCAO and vitro-thrombus, the rats were divided into the model(or control) group、positive group、crude drug group and treating group. Model(or control) group was given equal volume NS; positive group was given XiaoShuantong Tablet(included by Chinese pharmacopoeia); crude drug group was given dry extract of KangNaoxueshuan Tablet; treating group was divided into three groups (low、middle and high dosage).Every group was given drug by intragastric administration according volum.Results1. Visible neuromotor dysfunction has been emerged after cerebral ischemia in model group, and the symptom was aggravated when ischemia time was extended, neurological deficit score 3h, 2.80±1.3、6h, 3.00±1.33、24h, 3.56±1.5. Compared to model group, treating group could improve neurological symptom and the effect was dose- dependent, neurological deficit score was: low dosage, 3h, 1.44±0.53、6h, 1.56±0.53、24h, 1.67±0.87(P<0.01); middle dosage, 3h, 1.10±0.57、6h, 1.30±0.48、24h, 1.40±0.69(P<0.01); high dosage, 3h, 0.90±0.44、6h, 1.00±0.71、24h, 1.30±0.53(P<0.01).After 24h cerebral ischemia, infarce volume of model group was (27.51±4.71)%, and compared to model group , treating group could reduced infarct volume: low dosage, 3h, (15.37±7.21)% (P<0.01)、middle dosage, (14.60±7.00)%(P<0.01)、high dosage, (11.18±3.35)% (P<0.01).2. Treating group could reduce the dry and wet weight of vitro-thrombus, and the effect was dose -dependent. Compared to thrombus dry weight of model group: 4.93±2.17mg、low dosage: 3.35±1.44mg、middle dosage: 3.13±1.38mg(P<0.05); high dosage: 2.88±1.02mg(P<0.05).Thrombus wet weight of model group: 24.23±8.21mg、low dosage: 15.56±5.13mg(P<0.05), middle dosage: 12.26±3.55mg、(P<0.01), high dosage:11.08±3.10mg(P<0.01).3. Compared to control group, high dosage of KNT could impress platelet aggregation which induced by ADP, the 1、3、5 minute PAG and MPAG were: (40.24±12.9)%、(43.08±5.44)%、(56.43±7.93)% and (42.13±8.45)%(P<0.01). Middle dosage could reduce 1min PAG(P<0.05)and low dosage hadn’t obvious effect. Only high dosage could decrease blood platelets count(P<0.05), but middle and low dosage hadn’t obvious effect.4. Outstanding effect to HCG and PV wasn’t observed. But compared to control group, middle dosage could reduce (36.03±5.45)%、(41.41±7.71)% and (32.97±6.17)%(P<0.01)of normal WBV which in 37.5/s、90/s and 300/s shear rate respectively , and high dosage could reduce (30.31±11.44)%、(35.51±8.37)% and (19.84±3.80)%(P<0.05)of normal WBV.Conclsion The results presented here showed that KNT could improve neuromotor function and reduce infarct volume after cerebral ischemia, and displayed various effect against ischemia injury. The mechanism might be related to the effect on reducing WBV、repress platelet aggregation and interfer thrombogenesis.更多还原