【作者】 郭家娟；

【导师】 黄永生；

【作者基本信息】 长春中医药大学 ， 中医内科学， 2007， 博士

【摘要】 高血压病是我国常见病和引发心脑血管并发症最严重的疾病之一，中医药治疗高血压病已取得较大进展，但对中医药作用机制的研究尚需深入。夏膝口服液为导师黄永生教授治疗高血压肝肾失调、毒伤血络证的经验方，临床应用表明有较好的降压和改善临床症状的作用。前期的实验研究表明，本方能改善自发性高血压大鼠的血液流变学指标，改善血管内皮功能。本研究在前期研究的基础上，从中医学的整体观出发，从文献综述、理论研究及实验研究方面对其进行深入、系统地探讨。文献综述从古代和现代中医药对高血压病的病名、病因病机、证候学、证的客观化、中医药治疗等方面的研究情况进行了综述，并对当前在诊断及疗效评定标准、辨证分型和科研等方面存在的问题进行了述评和展望。从现代医学角度对高血压病发病过程中的“神经内分泌免疫调节网络”进行研究，并据此对近年来关注较多的神经递质、细胞因子、内分泌系统及其在高血压病病理过程中的相互调节作用进行了综述，并进行了评述和展望。理论研究在归纳、整理古今文献的基础上，论述了肝肾阴阳失调是高血压病的发病基础，毒伤血络是高血压病的发病关键；论证了在高血压病的治疗中滋阴潜阳与解毒通络的内在联系。提出调整肝肾、解毒通络是治疗高血压病的有效途径，夏膝口服液就是在此理论指导下创立的。该方在立意上，体现了调整肝肾阴阳盛衰的主旨，在组方上昭示了以解毒通络为先的特点，为中医药治疗高血压病提供了新的研究思路。实验研究40只12周龄雄性SHR随机分为SHR空白组、夏膝口服液组、卡托普利组、厄贝沙坦组，每组10只，另取10只WKY大鼠作为正常对照组，给药8周观察下述指标。1夏膝口服液对SHR心肾血管功能和结构的影响1．1对SHR行为和一般状态的影响研究表明夏膝口服液能改善SHR行为和一般状态，表明夏膝口服液对SHR中枢神经系统有一定的调理作用。1．2对SHR血压的影响夏膝口服液能抑制SHR血压随年龄增长而升高的趋势，给药4周后与空白组比较即有显著性差异（P＜0.05），且降压效果平稳持久。1．3对SHR左心室肥厚的影响夏膝口服液能降低SHR左心室重量指数（LVWI），与空白组比较有显著性差异（P＜0.01），表明夏膝口服液能在一定程度上逆转左心室肥厚（LVH）。1．4对靶器官损害的形态学观察：给药8周后，夏膝口服液能明显减轻SHR心肌细胞肥大，改善血管内膜状态，减轻肾脏间质纤维化，提示夏膝口服液能减轻靶器官损害。2夏膝口服液对神经内分泌免疫调节网络的影响2．1对儿茶酚胺的影响采用酶联免疫吸附法（ELISA）测定SHR血浆儿茶酚胺（CA）浓度。结果表明，SHR血浆CA浓度显著高于WKY对照组（P＜0.01）；灌胃给予夏膝口服液20天即可显著降低大鼠血浆CA浓度（P＜0.01），至第40天降至正常水平，与WKY对照组比较无显著性差异（P＞0.05），并稳定维持至第60天。揭示CA浓度升高可能是SHR神经兴奋、肝阳上亢及血压升高的物质基础，夏膝口服液能通过降低CA浓度而发挥对SHR神经系统的调理作用，起到降低血压及改善大鼠行为状态的作用。2．2对肿瘤坏死因子、细胞间粘附因子的影响采用放射免疫法（RIA）测定SHR血清肿瘤坏死因子（TNF-α）含量，用免疫组化法（ICH）检测血管内皮细胞间粘附因子-1（ICAM-1）蛋白表达。结果显示夏膝口服液具有降低血清TNF-α浓度，调节血管内皮ICAM-1蛋白表达的作用。揭示TNF-α及ICAM-1的高表达所引起的内皮损伤，可能是毒伤血络的客观病理基础；夏膝口服液能在降压的同时，通过调整TNF-α及ICAM-1的表达而实现对SHR免疫机制紊乱的调节作用。2．3对肾素-血管紧张素系统的影响肾素-血管紧张素系统（RAS）对心血管系统发挥着循环和局部体液调节的作用，本研究采用放免法测定血浆肾素活性（PRA）、血浆和心肌局部血管紧张素Ⅱ（AngⅡ）含量，用分光光度计检测血管紧张素转换酶（ACE）活性，用免疫组化法（ICH）检测心肌血管紧张素Ⅱ受体1型（AT₁R）的蛋白表达，用逆转录聚合酶链反应（RT-PCR）检测AT1RmRNA基因表达的含量。结果表明夏膝口服液对SHR具有降低PRA、血浆和心肌局部AngⅡ含量、ACE活性的作用，并降低心肌AT₁R蛋白及基因表达。揭示夏膝口服液可通过抑制肾素-血管紧张素系统活性，发挥全身性调节的功能而降低SHR血压，具有类似血管紧张素转换酶抑制剂（ACEI）及血管紧张素受体拮抗剂（ARB）的降压机制。结论SHR存在神经内分泌免疫调节功能的异常，夏膝口服液能通过多组分、多途径、多环节、多靶向的特点降低SHR血压，逆转左心室肥厚，改善靶器官损害，使SHR神经内分泌免疫调控网络恢复正常。更多还原

【Abstract】 In the present, hypertension has become one of the most common disease in China andhas often led to serious cardio-cerebral vascular complication. It has obtained great advancesin essential hypertension’s treatment by Chinese medicine. While the function mechanism ofChinese medicine need to be further investigated. Xia xi Oral Liquid is a proved recipe forhypertension, which is worked out by the author’s tutor Prof Huang Yong Sheng, deducingfrom the theory that the main mechanism of EH is the maladjustment of liver and kidney anddamage of superficial venues and lymph vessels. Clinical application has indicated that thisrecipe is quite effective in reducing blood pressure and improve clinical symptom. The furtherstudy indicates that the recipe could improve blood flowing deformation and ameliorateendothelium function of vascular in Spontaneously hypertensive rats （SHR）.Based on furtherstudy, this article been performed from the aspects of reference review, theory study as well asexperiment on SHR to observe its effects on neuron-endocrine-immune network.Reference reviewThe essay reviewed from the advances of such fields as followed: the ill-name, aetiologyand pathogenesis for EH, differential syndrome diagnosis, the substantial bases of thedifferential syndrome diagnosis, the treatment of the Chinese medicine, which is in ancientand modern times,moreover, included the evaluations and prospects on the criterions ofdiagnosis and curative effect as well as the problems lying in the selection of treatment basedon the differential syndrome diagnosis or scientific study. This essay summarized theexperimental study results and advances on the principle of neuron-endocrine-immunenetwdrk in EH according to such aspects as nervous intermediary, cell molecules, endocrinesystem and mutual regulation during the pathology process in EH. It also included evaluationsand prospects on these above.Theory studyBy inducing and sorting out the reference, we put forward the theory that the main bases of EH is the maladjustment of liver and kidney and the crux is the damage of superficialvenues and lymph vessels by poison, we expound and prove the inner link of adjustment ofliver and kidney and removing toxicity and obstruction, then put forward the theory that andremoving the toxicity and the obstruction is a valid path of treat Eli. Xia Xi Oral Liquid isoriginated from this theory which precipitating the point of adjustment of liver and kidney andthe characteristics of removing the toxicity and the obstruction, which provides theexperimental and theoretical bases for both its clinical application and its drug development.Experimental study40 12-week-old male SHR were divided randomly into SHR placebo group, Xia Xi OralLiquid control group, captopril control group and Irbesartanl control group, and 10 weightmatched male normotensive WKY rats were set as normal control（WKY group）.Observe suchaspects as followed after administration 8 weeks.1 The influence of Xia Xi Oral Liquid to the function and structure to cardiovascularsystem1.1 behavior and general condition effect: Xia Xi Oral Liquid can improve the behavior andgeneral condition in SHR. That indicates that Xia Xi Oral Liquid has regulation function tocenter nervous system.1.2 Decreasing blood effect: Xia Xi Oral Liquid can restrain the tendency that the blood willrise along with aging in SHR. It has a significant statistic sense between the Xia Xi OralLiquid treated group and the SHR placebo group after administration 4 weeks（P＜0.01）. XiaXi Oral Liquid can maintain drug effect smooth and steady.1.3 Reversing left ventricular hypertrophy（LVH） effect: Xia Xi Oral Liquid can reduce theratio of LVW to BW by 24.05%（P＜0.01）, decrease the left ventricular mass index.. It has asignificant statistic sense between the Xia Xi Oral Liquid treated group and the SHR placebogroup （P＜0.01）.Which indicates Xia Xi Oral Liquid can reverse LVH to a degree.1.4 Morphology observation on target organ damages: Xia Xi Oral Liquid obviouslyalleviates the hypertrophy of cardiac muscle, protect blood vessel endothelium and reduce theproliferation of smooth muscle in tunica media after administration 8 weeks. The resultsshowed Xia Xi Oral Liquid has the function of protecting to cardiovascular system in SHR.2 The influence of Xia Xi Oral Liquid to neuron-endocrine-immune network. 2.1 The influence of Xia Xi Oral Liquid to the density of catecholamine（CA） in plasmaStudying the density of CA in plasma in SHR with Enzyme-Linked ImmunosorbnentAssay （ELISA）. The results showed that the density of plasma CA in SHR is obviously higherthan that in WKY rats（P＜0.01）. Xia Xi Oral Liquid can reduce the lever of plasma CA inSHR, which had a significant statistic sense in comparing with placebo group afteradministration 20 days（P＜0.01）,and can reduce the lever to normal standard after 40 days（P＜0.01）, the tendency lasting for 60 days. The result showed that Xia Xi Oral Liquid has thefunction of reduce the density of CA in plasma in SHR. and increasing catecholaminestandard may be the inner basis for excitement, hyperactive liver-yang ascending andhypertension in SHR. Xia Xi Oral Liquid can depress the activity of sympathetic nerve, lowerblood pressure and improve behavior and general condition through reduce the density ofserum CA in SHR.2.2 The influence of Xia Xi Oral Liquid to Tumor necrosis factor-α（TNF-α） density inplasma and intercellular adhesion molecule-1 （ICAM-1） expression in kidney arteriole in SHRStudying the density of TNF-αin serum with radio-immunity（RIA）and ICAM-1expression in arteriole with immunohistochemistery （ICH）. The results showed that Xia XiOral Liquid can reduce TNF-αdensity in serum and ICAM-1 expression in arteriole in SHR.The results revealed that the injury of endothelial cell induced by higher expression in TNF-αand ICAM-1 may be one of the pathology basis about damage of superficial venues andlymph vessels by poison. And XiaXi Oral Liquid can alleviate the injury of endothelial cellthrough reduce ICAM-1 expression in SHR so that educe the function of removing toxicityand obstruction.2.3 The effect ofXia Xi Oral Liquid to rennin-angiotensin system （RAS）Studying the plasma rennin activity（PRA） and plasma angiotensinⅡ（AngⅡ） ontent withradio-immunity, serum Angi0tensin-Converting enzyme （ACE）activity withspectrophotometer, angiotensinⅡreceptor typeⅠ（AT₁R） expression in cardiac muscle withimmunohistochemistery （ICH） and the gene expressive content of AT₁R in cardiac musclewith Reverse Transcription -polymerasechain reaction（RT-PCR）in rats. The results showedthat Xia Xi Oral Liquid can reduce PRA, AngⅡcontent, ACE activity, AT1R expression andthe gene expressive content of AT₁R in cardiac muscle. It revealed that Xia Xi Oral Liquid has the function of lower blood pressure as Angiotensin-Converting enzyme inhibitor（ACEI）andangiotensin receptor block（ARB）.Conclusion: SHR occur the abnormity of neuron-endocrine-immune network. Xia XiOral Liquid could lower blood pressure, reverse left ventricular hypertrophy, and improve targetorgan damages through a multiple composition, multiple pathways, multiple links andmultiple targets. Recover the function of neuron-endocrine-immune network.更多还原