高血压病病机探讨及三草降压汤的降压作用与机理研究

【作者】 曲荣波；

【导师】 王庆国；

【作者基本信息】 北京中医药大学 ， 中医临床基础， 2007， 博士

【摘要】 高血压病是一种常见的慢性心血管疾病,不仅发病率高,而且具有病程长,病理机制复杂,并发症多等特点,常伴有心、脑、肾等重要脏器的损害,严重危胁着人类健康。因此,高血压病发病机理和治疗药物的研究,一直是国内外医学界研究的热点。越来越多的证据表明,中医药治疗高血压病,具有独特的优势。在长期的临床实践中,也积累了一些有效方剂。归纳分析高血压病的病机特点,系统评价临床有效方剂的疗效,阐明其降压机理,进而研制新的降压药物,为临床提供更有效的治疗方法,是中医药现代化的重要内容。病机在中医理论中占有十分重要的地位。本课题将高血压的病机按照不同层次,分别讨论,然后探讨各层次间的关系,融会贯通,以期理清病机系统的脉络。三草降压汤(SCD)是我校名老中医刘渡舟教授创制的经验方,经过大量临床观察证明对高血压病有效。本课题利用现代药理学的方法系统评价该方的降压作用,阐明其降压作用机理,以便为该方的开发和应用奠定现代研究的基础。利用“病证结合动物模型”进行中医药的基础和应用实验研究,所得研究结果方能揭示辨证论治、方证相应等中医理论的科学内涵,体现中医药治疗的特色与优势。本课题通过宏观表征和行为学指标的观察与微观指标的测定,较系统地研究了自发性高血压大鼠(SHR)和肾性高血压大鼠(RHR)的证候属性,为建立适合中医药研究的动物模型提供了新思路。目的1探讨高血压病的病机及SCD治疗高血压的组方机理;2评价SCD的降压作用;3阐明SCD治疗高血压病的作用机理;4初步辨析SHR与RHR模型的证候属性。方法1理论研究通过整理高血压病病机文献,较系统地探讨高血压病病机;基于高血压气、血、脉失调的病机,浅析SCD的组方特点。2实验研究采用SHR和两肾一夹(2K1C)法制作的RHR动物模型;采用清醒状态下大鼠尾动脉间接测压法观察降压作用的时效、量效关系及连续给药的降压效果;运用宏观表征和行为学指标,研究SCD对动物模型的证侯属性的影响;运用放射免疫、生物化学的方法检测方药对血浆中的血压调节物质的影响;运用病理切片的方法观察对靶器官损害的影响。结果1理论研究高血压病的病机可以分为基本病机,症状病机,疾病病机,证候病机及微观病机等几个不同层次,气、血失调和脉道不利是该病病机的关键。气、血、脉兼治是SCD的组方特点。2实验研究2.1 SCD对SHR大鼠的降压作用及其机理研究(1) SCD对SHR单次给药的时效关系研究表明,SCD小、中、大3个剂量组时效特点相似,均在给药后4h达到最大降压效果,而后开始回升,8h接近给药前水平。量效关系研究则表明给药后4h,中、大剂量组血压均显著下降(p<0.05),降幅达10mmHg左右,小剂量组无降压作用。(2) SHR模型组大鼠与正常组相比在14～18周内血压偏高(p<0.01),心率偏高(p<0.01),易激惹程度增加(p<0.05或p<0.01),痛阈提高(p<0.01),旋转时间偏低(p<0.01或p<0.05),眼球突出度偏高(p<0.01),体重偏低,18周龄时有显著性差异(p<0.05),神情较紧张,同笼大鼠间经常有打架现象,毛色发黄而无光泽,毛发脱落较多,小便颜色偏黄,大便颗粒较小、质地较干。连续给药4周期间,SCD组与模型组相比在给药后血压极显著下降(p<0.01),心率没有明显改变,易激惹程度降低(p<0.01),痛阈无明显差别,旋转时间增加(p<0.05)。眼球突出度未见显著改善,体重无显著变化,紧张、打架现象减少,小便颜色较清,大便较湿润。(3)病理形态上,SHR模型组可见心肌横径增粗;部分肾脏皮质可见肾小球硬化、萎缩,局部区域肾小球集中现象,间质中有淋巴细胞浸润;大脑皮层局部区域可见嗜神经细胞现象,脑软化等现象。连续给药4周的SCD组上述现象均较少见。(4)与正常组相比,SHR模型组心脏指数和左心室指数均增加(p<0.01),血浆AngII水平偏低(p<0.01),ET升高(p<0.01),ANP降低(p<0.01),PRA、CGRP、NO含量无明显差异。与SHR模型组相比,连续给药4周的SCD组的心脏指数和左心室指数无显著差异,血浆ET水平降低(p<0.01),ANP升高(p<0.01),NO升高(p<0.01),AngII、PRA、CGRP无明显变化。2.2 SCD对RHR大鼠的降压作用及其机理研究(1) SCD对RHR单次给药的时效关系研究表明,SCD小、中、大3个剂量组时效特点相似,均在给药后4h达到最大降压效果,而后开始回升,8h接近给药前水平。量效关系研究表明,给药后4h,小、中、大剂量组血压均显著下降(p<0.05或p<0.01),中剂量组降幅达17mmHg左右,大剂量组10mmHg,小剂量组6 mmHg。(2) RHR模型组大鼠在造模后4周～8周期间与正常组相比血压偏高(p<0.01),心率偏高(p<0.01或p<0.05),痛阈偏高(p<0.05),体重偏低,18周龄时有显著性差异(p<0.01),精神较萎靡,毛色无光泽,小便色清,被抓持时遗尿现象较普遍,大便质软,腐臭味重。连续给药4周期间,SCD组与模型组相比在给药后血压极显著下降(p<0.01),心率减慢(p<0.05),体重增加(p<0.05),痛阈增高(p<0.05),转台旋转时间增长(p<0.05),毛色不泽与遗尿现象亦有改善。(3)病理形态上,RHR模型组可见心肌横径增粗;部分肾脏皮质可见肾小球硬化、萎缩,局部区域肾小球集中现象,间质中有淋巴细胞浸润,球旁动脉可见玻璃样变性;大脑皮层局部区域可见嗜神经细胞现象,脑软化等现象。连续给药4周后,SCD组上述现象均较少见。(4)与正常组相比,RHR模型组心脏指数和左心室指数上均增加(p<0.01),血浆PRA水平偏高(P<0.01),AngII偏高(P<0.05),ANP偏高(p<0.01),ET、CGRP、NO无显著差异。与模型组相比,连续给药4周后,SCD组的心脏指数和左心室指数无显著差异,血浆PRA水平降低(p<0.01),AngII降低(p<0.05),ET降低(p<0.05),NO升高(p<0.05),ANP、CGRP无明显变化。2.3 SHR大鼠对SCD降压反应的个体差异初探虽然多数个体给予SCD后血压明显降低,但有29%的SHR大鼠在给药后血压无明显变化,当增大或减小剂量时,这些对药物不敏感的大鼠血压仍没有显著变化。结论1气血失调和脉道不利是高血压病病机的关键,气、血、脉兼治是SCD配伍特点;2 SCD具有明显的降压作用,并且其降压作用显示出一定时效、量效关系;3 SCD可以改善SHR和RHR的宏观表征及行为,显示出其整体治疗效果;4初步阐明了SCD的降压作用机理,发现SCD在两种高血压模型中作用机理不同;5早期SHR对SCD降压反应存在个体差异。更多还原

【Abstract】 Hypertension is a familiar chronic cardiovascular disease. Not only the incidence of hypertension is high, moreover, the course of the diseases is comparatively longer, its pathology mechanism is complex. This syndrome has many complications, that ofen associates with the damage of important viscera such as heart, brain, kidney and so on, which threatens the health of human being seriously. So the research on the pathogenesis of hypertension and curatives is one of the hotspots of international medical field all over the world. More and more proofs make it clear that T.C.M. has unique advantage on the therapy of hypertension and some available prescriptions have been accumulated in the long-term clinic practices. Reduction analysis of the pathology characteristics of hypertension, comprehensive evaluation the curative effect of clinic effective prescriptions, clarification the mechanism in treating , to study its mechanism in treating essential hypertension, thus development new therapy medicine of hypertension and providing more effective trerapy methods for clinic are important contents of modernization of T.C.M..Pathogenesis possesses very important postion in T.C.M. theory. This study discussed pathogenesis of essential hypertension respectively according to different levels, then probed into the relationship of various levels in order to ascertain the skeleton venation of pathogenesis system.San Cao Jiang Ya Tang (SCD) is an experience prescription formulated by famous old doctor of Chinese medicine Liu Du-zhou, a professor of our uviversity, the prescription has been proofed effective in treating hypertension. In this study we maked use of modern pharmacology, comprehensively evaluates this prescription’s effect on hypertension and clarifies its mechanism on therapy of hypertension so as to establish modern research foundation for the development and application of this prescription.Only through foundmental and applied experiment research by using“animal model with combined syndromes and symptoms”, can the research results reveal the scientific meanings of T.C.M. theories such as treatment based on syndrome differentiation, prescriptions corresponding to syndromes and so on, representing the characterisctis and advantages of traditional chinese medicine. This study comprehensively researched symptom attributes of the spontaneously hypertensive rat (SHR) and the renal hypertensive rat (RHR), providing new thinking on setting up modern animal models for T.C.M. researches.Objective 1 Discuss the mechanism of hypertension and SCD’s prescription theory in treating hypertension;2 Evaluate SCD’s effect in treating hypertension;3 Clarify SCD’s mechanism in treating hypertension;4 Differentiate Syndrome attributes between SHR and RHR elementarily.Methods1 Theoretics researchSystemically discuss the pathogenesis of hypertension through organizing literatures about it.Preliminary analysis the prescription theory of SCD based on maladjustment pathogenesis of Qi, blood and vessel in hypertension.2 Experiment researchSHR and two-kindy, one clip (2K1C) RHR were used as animal model. Tail-artery blood pressure measurement for conscious rats was used to observe the time-effect, dose-effect and long-term treatment features of the medicine to the blood pressure of the rats.The effect of SCD to animal model was studied by observeing macro representations and behavior. The blood pressure regulating factors in the plasma was examined by the method of radioimmunoassay and biochemistry. The target-organ damage was observed by pathologic slices making.Result1 Theoretics researchPathogenesis of hypertension can be divided into several different levels, such as basic pathogenesis, pathogenesis of representation, pathogenesis of disease, pathogenesis of syndromes, and micro pathogenesis. Maladjustment between Qi and blood and obstruct of the vessels are the key to the pathology mechanism of this disease. Prescription characteristics of SCD lies in that it can concurrently cure Qi, blood and vessels.2 Experiment research2.1 Research on SCD’s therapy on hypertension of SHR and mechanism(1) The time-effect and dose-effect study of one time treatment of SCD showed that, the time-effect feature of low, middle, high comcentration were similar, they all got the maximum antihypertensive effect at 4h after drug taken, then the blood pressure began to rise, and got back to the level before drug taken at 8h. The dose-effect study shows, the middle, high comcentration groups all had their blood pressure droped obviously at 4h aftert drug taken (p < 0.01), the blood pressure droped about 10mmHg, the low comcentration had no obvious antihypertensive effect.(2) Between 14-18 week-age, compared with the normal group, the SHR model group’s blood pressure was higher (p<0.01), heart rate was higher (p<0.01), irritable degree was higher (p<0.05 or p<0.01), pain threshold was higher (p<0.01), time persisting on rotary table was shorter (p<0.05 or p<0.01), eyes pop degree was higher (p<0.01), body weight was obviously lower at 18 week-age (p<0.05), and they looked more tentional, often fight between the cagemates, their hair color was more yellow, lacklusterits and losing more, their urine was more yellow, their dejection was more dry. In the 4 weeks treatment, compared with the model group, the SCD group’s blood pressure droped obviously (p<0.01), heart rate and pain threshold had no obvious change, irritable degree reduced (p<0.01), time persisting on rotary table extended (p<0.05), eye pop degree and body weight had no obvious chang, tension and fight reduced, urine color was ligter, dejection was wetter.(3) Pathomorphology shows, the transverse diameter of ventricular myocyte turned thicker; glomerular sclerosis, atrophy and centralize could be seen in some part of cortex of rats kidney, lymphocytic infiltration could be seen in thestromal; nerve cell addiction and encephalomalacia phenomenon could be seen in some part of the pallium. After 4 weeks treatment, the phenomena above were less in SCD group.(4) Compared with normal group, both heart weight index and left ventricular mass index of SHR model group inceased (p<0.01), blood plasma AngII level was lower (p<0.01), ET increased (p<0.01), ANP reduced (p<0.01), contents of PRA、CGRP、NO had no obvious differences.After 4 weeks treatment, compared with model group, the heart weight index and left ventricular mass index had no obvious difference, blood plasma ET level reduced (p<0.01), ANP hoisted (p<0.01), NO hoisted (p<0.01), AngII、PRA、CGRP had no obvious change.2.2 Research on SCD’s therapy on hypertension of RHR and mechanism(1) Time-effect study of one time treatment of SCD showed that, the time-effect feature of low, middle, high comcentration were similar, they all got the maximum antihypertensive effect at 4h after drug taken, then the blood pressure began to rise, and got back to the level before drug taken at 8h. The dose-effect study shows, the low, middle, high comcentration groups all had their blood pressure droped obviously 4h aftert drug taken (p<0.05 or p<0.01), the middle comcentration group’s blood pressure droped about 17mmHg, the high comcentration group’s droped 10mmHg, and the low comcentration group’s droped 6mmHg. (2) Compared with the normal group, the RHR model group’s blood pressure was higher (p<0.01), heart rate was higher (p<0.05 or p<0.01), pain threshold was higher (p<0.05), body weight was obviously lower at 18 weeks age (p<0.01), and it had lower spirits, its hair color was lackluster, its urine was more lighter-colored, it was enuresis being holded up, its dejection was more wet, and had a heavier moldy smell.In the 4 weeks being treated, compared with the model group, the SCD group’s blood pressure droped obviously (p<0.01), heart rate reduced, body weight hoisted, pain threshold hoisted, time persisting on rotary table extended, and the hair color and enuresis got better.(3) Pathomorphology shows, the transverse diameter of ventricular myocyte turned thicker; glomerular sclerosis, atrophy and centralize in some part of cortex of rats kidney, lymphocytic infiltration in thestromal and hyaline changes in the artery beside the glomerular could be seen; nerve cell addiction and encephalomalacia phenomenon can be seen in some part of the pallium. After 4 weeks treatment, the phenomena above were less in SCD group.(4) Compared with normal group, both heart weight index and left ventricular mass index of RHR model group inceased (p<0.01), blood plasma PRA level increased (p<0.01), AngII inctreased (p<0.05), ANP increased (p<0.01), contents of ET, CGRP, NO had no obvious differences.After 4 weeks treatment, compared with model group, the heart index and left ventricle had no obvious difference, blood plasma PRA level reduced (p<0.01), AngII reduced (p<0.05), ET reduced and NO hoisted (p<0.01), ANP, CGRP had no obvious change.2.3 Study on individual differences of blood pressure response of SHRs to SCD Although blood pressure decreased obviously in many individuals, there was no obvious difference in the blood pressure of 29% SHR after taken SCD. When the dose was increased or decreased, blood pressure of these insensitive rats still didn’t improve. Conclusion1 Maladjustment between Qi and blood and obstruct of the vessels are key factors of hypertension pathogenesis. Therapy on Qi,blood and vessels concurrently is SCD’S prescription characteristics;2 SCD has obvious effect on therapy of hypertension and the effect can last 6 hours, furthermore, its dosage and effect have some relativity;3 SCD can improve SHR and RHR’s syndrome, which shows its holistic therapy effects;4 the study illustrates SCD’s mechanism on the therapy of Hypertension mechanism elementarily and finds SCD has different operation mechanism on two hypertension models;5 In early stage of the disease, SHR’reaction on treatment of hypertension has individual difference.更多还原