脑络舒通胶囊对大鼠实验性脑及动静脉血栓

脑络舒通胶囊对大鼠实验性脑及动静脉血栓 形成的药效学研究 陕西咸阳抗衰老研究所 ,712000, 连秀峰 陈领朝 西安交通大学医学院药理学教研室 ,710061, 袁秉祥 孙晓东 于小江 提要:使用大鼠实验性脑血栓模型、动静脉血栓模型，观察脑络舒通胶囊对血栓形成作用的影响。 -1-1研究表明，脑络舒通胶囊大、中剂量(0.8 g/kg.d和0.4 g/kg.d)可抑制大鼠动静脉环路血栓的形成，使血栓湿重降低，血栓抑制率可达39,,50,，说明脑络舒通胶囊抗血栓作用明显;脑络舒通胶囊还可减轻脑栓塞引起的伊文思蓝蓝染和脑水肿，说明脑络舒通胶囊具有抑制脑血栓形成的作用。脑络舒通胶囊的上述药效学作用可能就是其驱风通络、逐瘀化痰、益气活血，治疗脑缺血、脑血栓急性期及恢复期等病证的药理学基础。 关键词:脑络舒通胶囊;脑血栓;动静脉血栓 脑络舒通胶囊是由地龙、当归、山楂、川芎各10g，水蛭、全虫各8 g，丹参、桃仁各12 g ，黄芪25 g等组成的纯中药复方制剂，具有驱风通络、逐瘀化痰、益气活血之功效，用于治疗脑缺血急性期及恢复期病症。本研究观察脑络舒通胶囊对大鼠实验性脑血栓模型及动静脉血栓模型血栓形成作用的影响。 1(实验材料 1(1 受试药品和试剂:脑络舒通胶囊，每粒0.4g，1.0g药粉相当于生药2.8g,，批号960815，由陕西咸阳抗衰老研究所提供。用生理盐水配成2,、4,和8,浓度的混悬液备用。偏瘫复原丸，北京同仁堂制药厂出品，批号9509118，用生理盐水配成8,的混悬液备用。尿激酶，山东蓬莱华泰制药有限公司出品，批号961020，用生理盐水配成2万u/ml，以备静脉注射用。 1(2 动物:SD大鼠58只，体重248?27g;雌雄各半，动物合格证陕医动05号，动物和饲料均由西安医科大学动物验实验中心提供。 〔1，2〕2(方法与结果 2(1 脑络舒通胶囊对大鼠动静脉血栓的影响 2(1(1 分组给药:将大鼠按体重和性别均衡随机分为6组， 除尿激酶静脉给药一次外，其它组均灌胃给药，灌胃容量均为每100g体重1ml，每天一次，连续三天，第四天制备动静脉环路血栓实验前1h再给药一次。?生理盐水对照组，灌胃给予生理盐水;?偏瘫the Communist Party of China, and a deep understanding of the party's nature, purpose, guiding principles, goals, organization, good style, deeply understand the conditions, rights and obligations of members, code of conduct, bearing in mind that oath, clear Members of the standards and conditions, enhance respect, abide by the Constitution, maintaining the party Constitution consciousness of the party Constitution. Read through the familiar with the code of self-discipline of the CPC, the CPC disciplinary regulations, members of the Communist Party of China party regulations on rights protection Ordinance, mastering self-discipline standards of "four must", the "four upholdings", mastering various types of disciplinary offence and punishment provisions and clear what you can do, what they can't do. 2. speech understanding learning series XI series the basic spirit of an important speech by General Secretary, learn to understand the Party Central Committee on governance of the new concept of basic elements of new ideas and new strategies, master and strengthen the party spirit, fulfills the purpose ideas, moral character, conservation related to the basic requirements. Learning of the XI series read the important speech by General Secretary (2016), XI Henan work research guide, General Secretary's important speech. Learning series speech, main understand master following aspects content: (1) ideal faith -1复原丸组，灌胃给予8,偏瘫复原丸混悬液，剂量为0.8g/kg.d;?尿激酶组，静脉给予尿激酶溶液(2万u/ml)每100g体重0.2ml，剂量为4万u/kg;?脑络舒通胶囊大、中、小 -1剂量组，分别给予2%、4%、8%的脑络舒通胶囊混悬液，剂量分别为0.2、0.4、0.8 g/kg.d。 2(1(2 制备动静脉环流血栓模型 〔〕1按参考文献方法，用3,戊巴比妥钠(30mg/kg)经腹腔麻醉大鼠，分离右侧颈总动脉及左侧颈外静脉。将两根肝素处理的聚乙烯管分别插入颈总动脉及颈外静脉，肝素生理盐水(50u/ml)充满聚乙烯管， 两管接口处放置一根5cm长的4号手术线。然后，开放动静脉环路15分钟，中断血流，迅速取出包裹丝线的血栓称重，并减去丝线重量，即得血栓湿重，并求出其血栓抑制率。 2(1(3 结果 以生理盐水组血栓湿重作为对照，进行非配对t检验，结果见表1。 表1. 脑络舒通胶囊对大鼠动静脉环路血栓的影响 x?s ?????????????????????????????? 组 别 例数 剂量 血栓湿重 血栓抑制率 (/kg) (mg) (%) ????????????????????????????? 生理盐水 10 20ml 19.54?5.88 偏瘫复原丸 10 0.8g 12.37?6.45* 36.7 尿激酶 8 4万u 9.65?3.95** 50.6 脑络舒通胶囊 小剂量 10 0.2g 20.80?10.20 -6.4 中剂量 10 0.4g 9.78?3.11** 49.9 大剂量 10 0.8g 11.93?4.28** 38.9 ????????????????????????????? 生理盐水为对比组 * P,0.05， ** P,0.01 -1-1研究表明，脑络舒通胶囊大、中剂量(0.8 g/kg.d和0.4 g/kg.d)可抑制大鼠动静脉环路血栓的形成，使血栓湿重降低，血栓抑制率可达39,,50,，说明脑络舒通胶囊抗血栓作用明显，但量效关系无显著意义(Tr检验，P,0.05)。 偏瘫复原丸也具有抑制血栓形成的作用，尿激酶具有血栓溶解作用。 2(2脑络舒通胶囊对大鼠实验性脑血栓的作用 2(2(1 药品和试剂 受试药品同前。 试剂:?凝血酶( Thrombase )， 南京比尔乐生化制药厂， 批号960610。?二磷酸腺苷(Arenosine diphosphate， ADP)，上海生物制品研究所，批号 960805。?肾上腺素(Adrenaline)，上海黄河制药厂， 批号 950104。?丙酮，西安化学试剂厂出品。? 血栓诱导剂:按100?200?5比例配备血栓诱导剂，ADP 1.25mmol/L;凝血酶 1.25万U/L;肾上腺素 1.0 g/L。 speech. Learning series speech, main understand master following aspects content: (1) ideal faithrtant f the XI series read the important speech by General Secretary (2016), XI Henan work research guide, General Secretary's impohen the party spirit, fulfills the purpose ideas, moral character, conservation related to the basic requirements. Learning orengtthe Party Central Committee on governance of the new concept of basic elements of new ideas and new strategies, master and st ech understanding learning series XI series the basic spirit of an important speech by General Secretary, learn to understandovisions and clear what you can do, what they can't do. 2. spediscipline standards of "four must", the "four upholdings", mastering various types of disciplinary offence and punishment pr-ina party regulations on rights protection Ordinance, mastering selfdiscipline of the CPC, the CPC disciplinary regulations, members of the Communist Party of Ch-h the familiar with the code of selfrespect, abide by the Constitution, maintaining the party Constitution consciousness of the party Constitution. Read throug nhanceghts and obligations of members, code of conduct, bearing in mind that oath, clear Members of the standards and conditions, eing of the party's nature, purpose, guiding principles, goals, organization, good style, deeply understand the conditions, rithe Communist Party of China, and a deep understand2 2(2(2 分组给药:将大鼠按体重和性别均衡随机分为6组， 除尿激酶静脉给药一次外，其它组均灌胃给药，灌胃容量均为每100g体重1ml，每天一次，连续三天，第四天制备脑血栓模型前1h再给药一次。?模型组，灌胃给予生理盐水;?偏瘫复原丸组，灌胃 -1给予8,偏瘫复原丸混悬液，剂量为0.8g/kg.d;?尿激酶组，灌胃给予生理盐水，造成脑血栓模型后静脉给予尿激酶溶液(2万u/ml)每100g体重0.2ml，剂量为4万u,kg; ?脑络舒通胶囊大、中、小剂量组，分别给予2,、4,、8,的脑络舒通胶囊混悬液,剂量分别 -1为0.2、0.4、0.8g/kg.d。 2(2(3 制备脑血栓模型 〔〕2按参考文献方法，用3,戊巴比妥钠(30mg/kg)经腹腔麻醉大鼠。分离气管，行气管插管术;分离右侧颈总动脉，顺颈总动脉向前分离右颈外动脉;结札颈外动脉和颈总动脉向心端，从颈总动脉向远心端插管备用。每只动物从颈总动脉注入血栓诱导剂每100g体重0.12ml。5min 后由右颈总动脉注射伊文氏蓝(0.2,)5ml/kg，5min后迅速断头处死，开颅取出大脑，分开左、右两半球，称取脑湿重。然后，将脑组织放入匀浆器中，加入0.5, SO4及丙酮(3?7)混合液5ml，制成匀浆， 移入试管密封静置60min。经3000r/minNa2 离心10min，取上清液，在620nm处测其光密度A值。以栓塞半球(右测为梗塞区，左测为对照区)的光密度A值与其脑重量之比表示伊文思蓝的含量，以此表示脑血栓的严重程度。以梗塞区半球重量与对照左半球重量的比值，表示梗塞侧脑水肿的程度。上述观察指标均以模型组为对比组，进行组间比较。 2(2(4 结 果 2(2(4(1 脑络舒通胶囊对大鼠脑梗塞区伊文思蓝含量的影响:结果见表2。 表2 脑络舒通胶囊对大鼠脑血栓伊文思蓝含量的影响 x?s ?????????????????????????????? 组 别 例数 剂量 A/脑半球重量 P (/kg) ????????????????????????????? 模型组 10 20ml 0.078?0.020 偏瘫复原丸 10 0.8g 0.043?0.017 ,0.01 尿激酶 8 4万u 0.026?0.012 ,0.01 脑络舒通胶囊 小剂量 10 0.2g 0.088?0.036 ,0.05 中剂量 10 0.4g 0.038?0.022 ,0.01 大剂量 10 0.8g 0.048?0.018 ,0.01 ????????????????????????????? 均与模型组比较 脑血栓模型制造后，肉眼可明显看出右测脑半球蓝染，以颅底蓝染程度为甚，左半球肉眼基本看不到蓝染，说明右测脑半球血栓模型制造成功。研究表明，脑络舒通胶囊中、 hl faitide, General Secretary's important speech. Learning series speech, main understand master following aspects content: (1) ideaasic requirements. Learning of the XI series read the important speech by General Secretary (2016), XI Henan work research guthe bnew strategies, master and strengthen the party spirit, fulfills the purpose ideas, moral character, conservation related to ecretary, learn to understand the Party Central Committee on governance of the new concept of basic elements of new ideas andral Scan do, what they can't do. 2. speech understanding learning series XI series the basic spirit of an important speech by Gene ur must", the "four upholdings", mastering various types of disciplinary offence and punishment provisions and clear what youdiscipline standards of "fo-protection Ordinance, mastering selfdiscipline of the CPC, the CPC disciplinary regulations, members of the Communist Party of China party regulations on rights -ty Constitution. Read through the familiar with the code of selfhe pare standards and conditions, enhance respect, abide by the Constitution, maintaining the party Constitution consciousness of tunderstand the conditions, rights and obligations of members, code of conduct, bearing in mind that oath, clear Members of thn, good style, deeply the Communist Party of China, and a deep understanding of the party's nature, purpose, guiding principles, goals, organizatio3 -1大剂量(0.4、0.8g/kg?d)可减少梗塞半球的伊文思蓝含量，说明其能减轻大鼠脑梗塞的严重程度。尿激酶作为临床常用药物对大鼠脑血栓模型有明显的实验治疗作用，偏瘫复原丸亦有一定的抗脑血栓作用。 2(2(4(2. 脑络舒通胶囊对大鼠脑血栓左右半球湿重的影响:结果见表3。 研究表明，脑血栓模型形成后，右脑湿重/左脑湿重的比例有所增加，脑络舒通胶囊大 -1剂量(0.8g/kg.d)可降低右脑湿重/左脑湿重的比例，并表现出一定的量效关系(Tr检验，P,0.01)， 说明脑络舒通胶囊可减轻脑栓塞引起的脑组织水肿。尿激酶表现出减轻脑水肿的趋势。偏瘫复原丸未表现出减轻栓塞性脑水肿的作用。 表3 .脑络舒通胶囊对大鼠脑梗塞侧脑组织水肿程度的影响x?s ?????????????????????????????? 组 别 例数 剂量 右/左脑半球重量 P (/kg) ????????????????????????????? 模型组 10 20ml 1.015?0.051 偏瘫复原丸 10 0.8g 0.995?0.032 ,0.05 尿激酶 8 4万u 0.983?0.028 ,0.05;,0.1 脑络舒通胶囊 小剂量 10 0.2g 1.040?0.051 ,0.05 中剂量 10 0.4g 0.997?0.054 ,0.05 大剂量 10 0.8g 0.964?0.040 ,0.05 ????????????????????????????? 均与模型组比较 3(讨 论 本研究成功造成了脑血栓模型，脑栓塞后，导致脑微循环障碍，小血管和毛细血管通透性增高，血脑屏障破坏，伊文思蓝通过血脑屏障进入脑实质，脑内伊文思蓝蓝染程度反映了脑血栓的严重程度。此外，脑血栓形成后，小血管和毛细血管通透性增高导致了脑水肿。研究表明，脑络舒通胶囊可减轻脑栓塞引起的伊文思蓝蓝染和脑水肿，说明脑络舒通胶囊具有抑制脑血栓形成的作用。 本试验用大鼠实验性脑血栓模型、动静脉血栓模型，观察脑络舒通胶囊对血栓形成作 -1-1用的影响。研究表明，脑络舒通胶囊大、中剂量(0.8 g/kg.d和0.4 g/kg.d)可抑制大鼠动静脉环路血栓的形成，使血栓湿重降低，血栓抑制率可达39,,50,，说明脑络舒通胶囊抗血栓作用明显;脑络舒通胶囊还可减轻脑栓塞引起的伊文思蓝蓝染和脑水肿，说明脑络舒通胶囊具有抑制脑血栓形成的作用。脑络舒通胶囊的上述药效学作用可能就是其驱风通络、逐瘀化痰、益气活血，治疗脑缺血、脑血栓急性期及恢复期等病证的药理学基础。 rtant f the XI series read the important speech by General Secretary (2016), XI Henan work research guide, General Secretary's impohen the party spirit, fulfills the purpose ideas, moral character, conservation related to the basic requirements. Learning orengtthe Party Central Committee on governance of the new concept of basic elements of new ideas and new strategies, master and st ech understanding learning series XI series the basic spirit of an important speech by General Secretary, learn to understandovisions and clear what you can do, what they can't do. 2. spediscipline standards of "four must", the "four upholdings", mastering various types of disciplinary offence and punishment pr-ina party regulations on rights protection Ordinance, mastering selfdiscipline of the CPC, the CPC disciplinary regulations, members of the Communist Party of Ch-h the familiar with the code of selfrespect, abide by the Constitution, maintaining the party Constitution consciousness of the party Constitution. Read throug nhanceghts and obligations of members, code of conduct, bearing in mind that oath, clear Members of the standards and conditions, eing of the party's nature, purpose, guiding principles, goals, organization, good style, deeply understand the conditions, rithe Communist Party of China, and a deep understandspeech. Learning series speech, main understand master following aspects content: (1) ideal faith4 主要参考文献 [1]陈奇主编.中药药理研究方法学.北京，人民卫生出版社，1993;571,572 [2]陈奇主编.中药药理研究方法学.北京，人民卫生出版社，1993;564 hl faitide, General Secretary's important speech. Learning series speech, main understand master following aspects content: (1) ideaasic requirements. Learning of the XI series read the important speech by General Secretary (2016), XI Henan work research guthe bnew strategies, master and strengthen the party spirit, fulfills the purpose ideas, moral character, conservation related to ecretary, learn to understand the Party Central Committee on governance of the new concept of basic elements of new ideas andral Scan do, what they can't do. 2. speech understanding learning series XI series the basic spirit of an important speech by Gene ur must", the "four upholdings", mastering various types of disciplinary offence and punishment provisions and clear what youdiscipline standards of "fo-protection Ordinance, mastering selfdiscipline of the CPC, the CPC disciplinary regulations, members of the Communist Party of China party regulations on rights -ty Constitution. Read through the familiar with the code of selfhe pare standards and conditions, enhance respect, abide by the Constitution, maintaining the party Constitution consciousness of tunderstand the conditions, rights and obligations of members, code of conduct, bearing in mind that oath, clear Members of thn, good style, deeply the Communist Party of China, and a deep understanding of the party's nature, purpose, guiding principles, goals, organizatio5