节点文献
加味大柴胡汤抗家兔实验性动脉粥样硬化作用机制的实验研究
【作者】 王凤荣；
【导师】 李敬林；
【作者基本信息】 辽宁中医药大学 ， 中西医结合临床， 2007， 博士
【摘要】 动脉粥样硬化(AS)是多病因的疾病，其主要危险因素有：血脂异常、高血压、吸烟、糖尿病等。近年来，随着生活水平的提高，本病发生呈上升趋势。AS所致的心脑血管疾病严重威胁人类的健康。本论文通过动物试验，从细胞分子水平对中药加味大柴胡汤防治AS的机理进行了较为深入的探讨，并对中西医防治AS的进展进行了较为全面的概述。实验研究：目的采用高脂饲料喂养家兔造成实验性动脉粥样硬化模型，观察加味大柴胡汤对家兔实验性AS的形成、对血管壁平滑肌细胞表型及相关原癌基因c-myc表达的影响，以及与AS炎症反应相关细胞因子主动脉单核细胞趋化蛋白-1(MCP-1)、主动脉抗原CD40-CD40L、细胞间黏附分子-1(ICAM-1)表达的影响。方法：本实验分正常对照组，AS模型对照组，加味大柴胡汤预防组，加味大柴胡汤治疗组和血脂康阳性对照组。治疗组造膜6周后开始给药，其余各组持续喂养10周，观察加味大柴胡汤对实验性AS家兔的血脂、氧自由基、C反应蛋白(CRP)的影响。通过光镜、电镜观察各组主动脉的病理形态学变化，观察此药对血管壁平滑肌细胞表型的影响，采用原位杂交方法，观察对相关原癌基因c-myc表达的影响。采用逆转录聚合酶链式反应(RT-PCR法)蛋白质印迹杂交(western Blot)及免疫组化法，进一步观察加味大柴胡汤对实验性AS家兔炎症反应相关细胞因子主动脉单核细胞趋化因子—1(MCP-1)、主动脉抗原CD40、CD40L、ICAM-1mRNA及蛋白表达的影响，探讨此药抗AS的作用机制。结果显示：加味大柴胡汤能降低胆固醇、甘油三酯、低密度脂蛋白胆固醇及脂质过氧化酶；提高超氧化物歧化酶、磷脂氢谷胱甘肽过氧化物酶；抑制原癌基因的表达。尤其加味大柴胡汤预防组降低血清CRP的水平优于治疗组和血脂康组，有显著性差异P＜0.05、P＜0.01。对MCP-1、ICAM-1表达的影响，加味大柴胡汤预防组与治疗组、血脂康组比较明显降低，经统计学处理P＜0.01，有极显著性差异；。说明加味大柴胡汤能降低MCP-1、ICAM-1的表达，以早期用药效果显著。对CD40、CD40L表达的影响，加味大柴胡汤预防组与治疗组比较明显降低P＜0.01，预防组与血脂康组比较无显著性差异。结论：加味大柴胡汤具有以下几方面作用①具有明显的调节脂质代谢，抗脂质过氧化损伤的作用；②阻断主动脉内膜的增生，抑制血管壁平滑肌细胞表型的改变；③阻抑原癌基因的表达；④降低血清CRP水平；⑤抑制实验性家兔主动脉内膜AS斑块处炎性因子MCP-1、ICAM-1、CD40、CD40L的表达，从而抑制主动脉内膜的炎症反应，防止AS的产生。因此，加味大柴胡汤防治AS的机理与调节血脂，抗脂质过氧化损伤，抑制血管壁平滑肌细胞表型的改变，阻抑原癌基因c-myc的表达，减少主动脉炎性因子的表达等有关。该实验研究结果将为加味大柴胡汤进一步的临床推广应用提供理论依据。理论研究以AS从痰瘀论治及炎性因子与AS的关系二方面进行阐述。具体包括“高脂血症的防治与中医养生学”、“冠心病从痰瘀论治”、“CD40与AS的关系”、“单核细胞趋化因子与AS的关系”、“AS现代医学研究进展”、“AS的中医药研究概况”。在理论研究及文献综述中，参照古今中医有关文献对动脉粥样硬化的中医病名、病因病机、辨证治疗方法、临床及实验研究进展和中医研究中存在的问题及对策进行了探讨。强调从痰瘀论治动脉粥样硬化，并对现代医学在动脉粥样硬化研究中所取得的进展进行了总结，进一步明确了动脉粥样硬化的病理特征、发病机制和治疗原则。并具体探讨了现代医学关于炎性因子(主动脉抗原CD40-CD40L和单核细胞趋化因子—1)在动脉粥样硬化发病机制中的作用。
【Abstract】 Background: Atherosclerosis (AS) is a disease which is characterized by many theories.Major riskfactors of AS are associated with abnormal blood lipid levels,hypertension,cigarettesmoking,diabetes.In the past few years, the mortality rate for AS has markedly and progressivelyincreased in China. AS also causes a great deal of serious illness which included coronary arterydisease (CAD), cerebrovascular disease.This article reviewed the advancement of traditionalChinese and modem medicine about AS.Objective: To investigate the mechanism of changed chinese herbal medicine Da-Chai-Hu-Tang ininhibiting AS in rabbits, observing the effects of Da-Chai-Hu-Tang for the formation of AS,theexpression of phenotype of smooth muscle cell of vessel wall and c-myc, monocyte chemoattractantprotein-1 (MCP-1),CD40,intercellular adhesion molecule (ICAM-1) in AS.Methods: The models of AS were set up by feeding containing hyperlipemia.The rabbits weredivided randomly into five groups:normal control group,AS model group,Da-Chai-Hu-Tangprevention group and cure group,xuezhikang positive control group. Da-Chai-Hu-Tang cure groupwas administered at the 6th week after models was set up,the other groups were feeded for 10weeks, observing the effect of Da-Chai-Hu-Tang for blood lipid,oxygen free radical,cAMP reactiveprotein. After sacrificing the animals, differential parts were obsevered, viewing aorticpathomorphology by light microscope and electron microscope and observing cellular phenotypeson smooth muscle cell and researching the effect for the phaenotype of smooth muscle cell of vesselwall and the expression of c-myc by hybridization in situ, studing MCP-1,CD40,ICAM-1,mRNAand expression of protein by reverse transcriptase PCR (RT-PCR),Westem Blot andimmunohistochemistry.Results: The levels of total cholesterol(TC),triglyeride(TG),low density lipoproteincholesterol(LDL-C) in serum were significantly lower and the activity of SOD,phospholipidhydroperoxide glutathione peroxidase (PHGPx) was improved in Da-Chai-Hu-Tang group.Thedegree of CRP was lowered Da-Chai-Hu-Tang prevention group has an advantage overDa-Chai-Hu-Tang cure group and xuezhikang positive control group.There was significantdifference (P＜0.05,P＜0.01).The expression of MCP-1 and ICAM-1 was less in Da-Chai-Hu-Tangprevention group than Da-Chai-Hu-Tang cure group and xuezhikang positive control group(P＜0.01).For the effect of the expression of CD40 and CL40L, there was significant difference betweenDa-Chai-Hu-Tang prevention group and cure group(P＜0.01), while there was no significantdifference between Da-Chai-Hu-Tang prevention group and xuezhikang positive control group.Conclusion: The prevention and cure mechanism of Da-Chai-Hu-Tang might be concerned withadjusting blood lipid,preventing the process of lipid peroxidation, inhibiting the phenotypic changeof smooth muscle cell and the expression of c-myc and inflammatory factor.This study providedthe evidence of theroy for the application of Da-Chai-Hu-Tang.The relation of phlegmatic stagnation and inflammatory factor was explained in study oftheory. Discussing hyperlipemia from phlegmatic stagnation,the prevention and cure ofhyperlipemia ,the study of health cultivation of Chinese medical science, CAD from phlegmatic,the relation of CD40 and AS,monocyte chemotactic factor and AS,the advancement of research for ASin modem medicine, the overview of Chinese medicine for AS,which were included in the article.Itwas effective in discussing AS from phlegmatic stagnation, investigating the name of illness,causeof disease,differentiation of symptoms and signs, treatment, some problems in study.it was indicatedfurther that the character of pathology, pathogenesy and therapeutic principle of AS, investigatingthe effect of inflammatory factor, CD40 and MCP-1 in AS.