【作者】 孙忠；

【导师】 彭康；

【作者基本信息】 第一军医大学 ， 中药学， 2004， 硕士

【摘要】 目的：在对补阳还五汤理论研究的基础上，方证结合，通过复制中风后遗症“气虚血瘀”大鼠病证模型，探讨该方主治中风后遗症的药理作用和相关机制，从而为该方的临床应用和深入研究提供依据和线索。 方法：补阳还五汤用传统方法煎制，黄芪甲苷（Astragaloside Ⅳ）为该方补气部分的代表成分，阿魏酸（Ferulic acid）为活血部分的代表成分，以二者共同作为该方的质量控制标准。黄芪甲苷的含量采用高效液相色谱—蒸发光散射检测器（HPLC—ELSD）测定；阿魏酸的含量采用高效液相色谱—紫外检测器（ELSD）测定。中风后遗症“气虚血瘀”大鼠病证模型的复制采用多因素复合法制作，以其行为变化、神经功能缺失评分、“气虚血瘀”外在表现、脑组织梗死体积、光镜和电镜下病理等作为该模型成立的判定指标。实验分正常组、模型组、低剂量组（12.5g／kg）、中剂量组（25g／kg）、高剂量组（50g／kg）和梗死率判断组等6组，大鼠灌胃给药15天后腹主动脉取血，用血液流变性测定仪检测全血粘度、血浆粘度、红细胞变形指数、红细胞压积、血小板聚集率等；取脑组织，采用原子分光光度法测定Na⁺，K⁺，Ca²⁺，Mg²⁺的含量，采用高效液相色谱法测定ATP，ADP以及NE，DA和5-HT的含量。 结果： （1） 黄芪甲苷在1.08μg～12.96μg（r=0.9995）范围内线性关系良好，阿魏酸在0.4μg～4.0μg（r=0.9995）范围内线性关系良好。 （2） 术后35天，各给药组大鼠神经功能缺失评分均显著低于20天评分印<0.01);低、中剂量组大鼠肢体偏瘫症状改善明显，饮食和饮水增加，体质量增加，行动相对敏捷，毛发较以前有光泽;高剂量组肢体偏瘫症状亦有好转，但精神略显不振且大便塘;模型组肢体偏瘫症状改善不明显，伤口愈合较慢。 （3）血液流变性方面，除高剂量组对55-1、305一，切变率时的全血粘度与模型组比较无统计学意义（夕0.05）外，其余各项指标（包括全血粘度、血浆粘度、红细胞变形指数、红细胞压积）均显著降低（p<0.001一0.05）;血小板聚集率各剂量组与模型组比较，均显著降低印<0 .001⁰.05)，高剂量组显著低于中剂量组印<0 .05)。 （4）离子代谢方面，模型组Na+显著高于正常组（p<0.01），中剂量组Na+显著低于模型组（p<0.05）:中剂量组oaz+显著低于模型组（p<0 .01）;中剂量组M扩+显著高于模型组（p<0 .01）。 （5）能量物质代谢方面，户JP含量模型组显著低于正常组印<0.001)，各给药组显著高于模型组印<0 .001⁰.01);ADP含量模型组显著低于正常组印<0 .001)，各给药组显著高于模型组印<0 .001)。 （6）单胺类神经递质方面，NE含量模型组显著低于正常组印<0 .001)，各给药组显著高于模型组印<0.05⁰.01);DA含量模型组显著低于正常组印<0.001)，各给药组显著高于模型组印<0.05⁰.01);5一HT含量模型组显著低于正常组印<0.001)，中剂量组显著高于模型组勿<0 .05)。 结论:本实验采用黄蔑甲昔和阿魏酸的检测技术准确、快速、灵敏，可以作为补阳还五汤质量控制标准的测定方法:补阳还五汤对中风后遗症“气虚血癖”大鼠病证模型有显著的治疗作用，明显改善模型大鼠肢体偏瘫症状，使其体质量增加，毛发润泽，食欲改善，活动增强，伤口愈合加快等，从而减轻模型大鼠“气虚血癖”外在表现;可纠正其血液流变性异常，降低血液粘度、抑制红细胞聚集并改善其变形性、抑制血小板聚集，起到明显的活血化癖作用:对脑细胞中Na+，G扩十的升高和K十，M犷+的降低有对抗和治疗作用，并可促进ATp，ADP代谢的恢复，有效改善能量代谢;对脑组织中单胺类神经递质NE，DA及5一HT的降低有对抗和纠正作用，从而有效促进神经功能恢复。 本研究的特点主要体现在:（1）结合方剂的配伍规律确定其质量控制成分，本研究以补阳还五汤君药黄茂的主要有效成分黄茂甲普作为补气部分的代表成分，以臣药当归及佐药川芍的阿魏酸作为活血部分的代表成分，共同作为质量控制标准，保证了实验的稳定性，也充分体现了中药复方质量控制标准制定的特色。（2）方证结合，依据中医方剂学基本理论，采用多因素复合方法复制了与补阳还五汤主治证相适应的中风后遗症“气虚血癖”动物病证模型。能较准确地反映该方的基本作用内涵。（3）选择血液流变性变化作为补阳还五汤活血作用的主要指标，脑组织离子Na+，K+，c扩十，Mg2+和ATP，ADP变化作为反映该方补气改善能量代谢的主要指标，以脑组织单胺类神经递质变化作为该方补气活血促进神经功能恢复的主要指标，在一定程度上揭示了补阳还五汤补气活血作用的现代药理机制。更多还原

【Abstract】 Objective: On the base of theoretic studies of BYHWD, combining prescriptions and syndromes and through replicating the stroke sequelae models of rats manifested as syndrome of qi deficiency and blood stasis,we studied the pharmacology effects and action mechanisms of BYHWD on treatment the stroke sequelae models of rats in order to offer the bases for clinic and the clues for the further studies.Methods: BYHWD was processed with traditional methods; Astragaloside IV was the main composition of supplying gas and ferulic acid was the main composition of promoting blood circulation of BYHWD.We selected astragaloside IV and ferulic acid as the quality control standards together.Detected astragaloside IV with the eveaporative light scattering detector and ferulic acid with UV one of high performance liquid chromatography;Replicated the stroke sequelae models of rats manifested as syndrome of qi deficiency and blood stasis. We seleceted the behavior changes,the deficiency grades of nerval founctions,the exterior representations of qi deficiency and blood stasis.the infarct volume of brains and the pathology changes observed from light and electricity lens as the basal indexes of determinant the models coming into existence. Grouped 72 Wistar rats into the nomal group,the control group.the low dosagegroup(12.5g/kg),the middle dosage group(25g/kg),the high dosage group(50g/kg) and the infarct ratio estimated group ransdomly.The rats were treated with BYHWD and 15 days later they were killed.Adopted atom spectrophotometer to assay the Na+,K+,Ca2+,Mg2+ content in the brains of the models,adopted high performance liquid chromatography methods to assay the ATP,ADP and monoamin neurotransmitters(NE,DA,5-HT)content in the brains of the models, and adopted correlative methods to determine such indexes as the whole blood viscosity,blood plasma viscosity,RBC deforrnability,RBC and platelet aggregation etc. which content in the blood of the models. Results:(1) The good linear correlation of astragaloside IV was observed from 1.08 u g to 12.96 u g(r =0.9995)and that of ferulic acid was observed from 0.4 n g to 4.0 u g(r =0.9995).(2) There were significant differences(p<0.01) in each BYHWD treated group about the deficiency grades of nerval founctions at 35d compared with those of 20d.The body hemiplegy syndromes of the middle and low dosage groups were improved greatly. Their weighth were increased,their hair relative lenitive and they acted smartly.The body hemiplegy symptoms of the high dosage group were recovered too but they were relative depressed and their stool was watery.The body hemiplegy symptoms of the control group were not reformed obviously and the wounds were indolence.(3) There were significant differences(p<0.001~0.05)in every other index except the high dosage group having no marked effects on the whole blood viscosity at 5s"1 30 s"1. Compared with the normal group there were significant differences(p<0.001 ~ 0.05)in ADP induced blood platelet congregation in each group and compared with the middle dosage group,there was significant difference(p<0.05)in the high dosage group of it.(4) Compared with the control group,there were significant differences(p<0.05~0.01)in Na+,Ca2+and Mg2+contented in the brains of themiddle dosage and compared with the normal group there was significant difference(p<0.05)in Na+ contented in the brains of the control group.(5) Compared with the nomal group, there were significant differences(p<0.001)in ATP and ADP contented in the brains of the control group and compared with the control group there were significant differences(p<0.001~0.01)in ATP and ADP contented in the brains of each treated group.(6) Compared with the nomal group,there were significant differences(p<0.001)in NE.DA and 5-HT contented in the brains of the control group and compared with the control group there were significant differences(p<0.05~0.01)in NE,DA and 5-HT contented in the brain of each treated g更多还原