【作者】 张京春；

【导师】 张文高； 陈可冀；

【作者基本信息】 山东中医药大学 ， 中西医结合临床， 2007， 博士

【摘要】 目的:研讨陈可冀院士活血化瘀治疗冠心病(CHD)的经验,探讨动脉粥样硬化(As)易损斑块的瘀毒病机及解毒活血配伍方干预的理论基础;观察解毒活血配伍方对载脂蛋白E基因敲除[ApoE(-/-)]小鼠As及其易损斑块的影响,探讨其作用机理。方法:通过基于病案数据库的数据挖掘与整理分析,总结陈可冀院士运用活血化瘀方法治疗CHD学术经验;在此基础上结合中西医理论与实践对易损斑块的瘀毒病机及其解毒活血治法进行理论探讨。将13周龄ApoE(-/-)小鼠分为高脂饲料模型组(98只)、普通饲料模型组(12只),同时设给予普通饲料的13周龄C57 BL/6J小鼠正常对照组(12只)。饲养19周随机抽取高脂饲料组ApoE(-/-)小鼠2只确认易损斑块形成后,随机分为8组:洛伐他汀组(灌服人用量10倍的洛伐他汀),血脂康组(灌服人用量10倍的血脂康),解毒组(灌服预计人用量10倍的虎杖提取物),活血组(灌服人用量10倍的芎芍胶囊),解毒活血配伍低剂量组(灌服人用量5倍的虎杖提取物及芎芍胶囊),解毒活血配伍中剂量组(灌服人用量10倍的虎杖提取物及芎芍胶囊),解毒活血配伍高剂量组(灌服人用量20倍的虎杖提取物及芎芍胶囊);高脂饲料模型组、普通饲料模型组与正常对照组小鼠均灌服生理盐水。连续灌胃给药17周,腹腔麻醉后下腔静脉取血检测血脂水平(TG、TC、LDL-C、HDL-C、VLDL),ELISA方法检测超敏C反应蛋白(hs–CRP)、单核细胞趋化蛋白-1(MCP-1)及白细胞分化抗原40配体(CD40L);取主动脉,HE染色光镜观察,透射电镜观察;免疫组化方法检测主动脉标本核因子-κB (NF-κB)表达及基质金属蛋白酶-9(MMP-9)表达。结果:1.基于病案数据库的数据挖掘分析的结果,明确了陈老师在冠心病血瘀证的病证结合辨证、不同活血化瘀治法及具体方药应用等方面的丰富经验,其在血府逐瘀汤基础上简化方药创制的芎芍胶囊为本课题的重要研究基础。2.从中医理论与中西医结合角度探讨,提出易损斑块瘀毒病机的创新思维,解毒活血治法的新思路,及体现本治法的虎杖配伍芎芍胶囊的新方案。3.实验结果显示:(1)解毒活血配伍方能调节ApoE(-/-)小鼠的血脂水平;(2)解毒活血配伍方能显著降低ApoE(-/-)小鼠血液hs-CRP、MCP-1、CD40L等炎性因子的水平;(3)解毒活血配伍方能显著降低ApoE(-/-)小鼠主动脉NF-κB和MMP-9表达;(4)解毒活血配伍方能抗SMC增殖、抗As斑块形成,保护主动脉的形态结构特别是超微结构,稳定斑块。解毒活血配伍方高剂量组对各项指标的影响优于中、低剂量组,某些指标优于洛伐他汀或血脂康对照组;解毒活血配伍方对NF-κB和MMP-9表达的影响优于解毒组与活血组。结论:陈老师活血化瘀治疗CHD的经验有重要的临床指导意义和学术价值。易损斑块及其作为病理基础的急性冠脉综合征(ACS)的瘀毒病机、解毒活血治法及其具体方案(虎杖配伍芎芍胶囊)的提出,具有理论和实际指导价值。解毒活血配伍方具有确切的调脂、抗炎、抗As与稳定易损斑块等作用,优于单纯解毒或活血,可以作为稳定As斑块、防治ACS的有效中医药治法和干预措施,其作用机理可能与调脂、抗炎、抗As、抑制基质降解等有关。更多还原

【Abstract】 Objective: To study and analysis the experience of academician Chen Ke-ji in treating coronary heart disease by means of activating blood to remove stasis, and discuss the mechanism of vulnerable plaque and the theoretical basis of the intervention with toxin-resolving and blood-activating. To observe the efficacy of combination with toxin-resolving and blood-activating on the atherosclerotic vulnerable plaque in apolipoprotein E gene knockout ( ApoE(-/-) ) mice and discuss its mechanism.Methods: Through studying and analyzing the cases data-base, to summarize the academic experience in treating CHD by means of activating blood to remove stasis, and discuss the mechanism of“toxin and stasis”and the intervention of combination with toxin-resolving and blood-activating on atherosclerotic vulnerable plaque. 110 ApoE (-/-) mice of 13-week old were assigned into two groups, 98 mice in Group A which were fed with hyperlipidemic diet, 12 mice in Group B which is fed with normal diet. Besides, a normal control group was set up with 12 C57BL/6J mice. After 19 weeks, 2 mice in Group A were randomly collected to confirm the formation of vulnerable plaque, the remaining in the group was divided into 8 subgroups: Lovastatin group was fed with lovastatin of 3.3 mg/kg.d as one dose (ten times than that for human-being of 0.33 mg/kg.d); Xuezhikang group was fed with Xuezhikang of 0.2 mg/kg.d as one dose (ten times than that for human being of 0.02 mg/kg.d);Toxin-resolving group was fed with polydatin (PD) of 26.6 mg/kg.d as one dose (ten times than that for human- being of 2.66 mg/kg.d); Blood- activating group was fed with Xiaongshao capsule of 110 mg/kg.d as one dose (ten times than that for human being of 11mg/kg.d); group in low dose was fed with PD of 13.3 mg/kg.d and Xiongshao capsule of 55mg/kg.d as one dose (5 times than that for human being’s); Combination of toxin- resolving and blood-activating group in medium dose was fed with PD of 26.6 mg/kg.d and Xiongshao capsule of 110mg/kg.d as one dose(10 times than that for human being’s); Combination of toxin-resolving and blood-activating group in high dose was fed with PD of 52.2 mg/kg.d and Xiongshao capsule of 220mg/kg.d as one dose (20 times than that for human-being’s); Normal saline of 0.4ml/d was given to mice in hyperlipidemia model group, normal diet group as well as normal control group. Forage at SPF level and PH2.8-3.0 acid water was provided to the above mentioned 10 groups of mice. After 17 weeks of intervention, to observe the effect on TC, TG, LDL-C, HDL-C and VLDL. Hs-CRP, MCP-1 and CD40L were detected with ELISA method. To take aorta and take pathomorphology observation and assay the expression of NF-κB, MMP-9.Results: 1. It manifests that Professor Chen is superior in treating CHD with activa- ting blood to remove stasis method, and Xiongshao Capusle that comes from Xuefuzhuyu Compound Formula is an important basis of this study. 2. Bring forward the toxin and sta- sis mechanism of vulnerable plaque, and combination of toxin-resolving and blood- activating is one of new methods in treating vulnerable plaque. 3. Experiments shows that (1)Combination of toxin-resolving and blood-activating herbs could reduce blood lipid level of ApoE(-/-) mice; (2)Combination of toxin-resolving and blood- activating herbs could reduce hs-CRP、MCP-1 and CD40L level in serum; (3)Combination of toxin-resolving and blood-activating herbs could reduce the expression of NF-κB and MMP-9 in aorta of ApoE(-/-) mice;(4)Combination of toxin-resolving and blood- activating herbs has the effects for anti-proliferation of SMC, anti-athrosclerotic plaque formation, protecting configuration structure of aorta, and stabilizing plaque. Combination of toxin-resolving and blood-activating herbs in high dose is superior to medium and low dose, and is superior to lovastatin group, Xuezhikang group, toxin-resolving group and blood-activating group in some indexes.Conclusion: The experience of Professor Chen in treating CHD with activating blood to remove stasis method has important clinical and academic significance. Toxin and Stasis is an important mechanism of vulnerable plaque and acute coronary syndrome (ACS) in Chinese medicine. Combination of toxin-resolving and blood-activating herbs possess the exact effects for lipid-reducing, anti-inflammation, anti- atherosclerosis, stabilizing vulnerable plaque, and is superior to individual toxin-resolving or blood-activating herb. It could be one of the effective TCM methods in treating and preventing atherosclerosis, stabilizing atherosclerotic plaque and ACS.更多还原