【作者】 苗治国；

【导师】 陈宝田；

【作者基本信息】 南方医科大学 ， 中西医结合临床， 2010， 博士

【摘要】 背景在我国,随着社会生活水平的提高以及生活方式的转变,高血压病的发病率稳步上升,2002年全国居民营养与健康状况调查资料显示,我国成人高血压患病率为18.8%,全国有高血压患者约1.6亿。作为最常见的慢性病,高血压是引起脑卒中、冠心病和肾功能衰竭等疾病的重要基础疾病之一。西药降压虽然具有降压迅速、顺应性良好的优势,但降压药的不良反应较多,很多病人甚至不能忍受而放弃治疗。中医药治疗高血压具有独特的优势,如降压稳定而持久,同时症状改善明显,不良反应较少。陈宝田教授根据祖国医学中“诸风掉眩,皆属于肝”、“无痰不做眩”等相关理论,结合数十年临床经验,创制加味镇眩方,用于高血压病的治疗,效果良好。如能合理开发应用,将有助于弥补西药降压的不足,平稳降压、改善高血压患者临床症状、减少降压药物的不良反应,提高我国近两亿高血压患者的生活质量和健康水平。在高血压的病理生理中,微血管对高血压及其并发症的形成可能具有更重要的作用。微血管密度的减少,不仅导致血液平行通路下降,而且引起外周血管阻力的增高,从而推动动脉血压的持续升高；微血管密度的减少,又能使各个靶器官组织的物质能量代谢水平、储备能力及物质交换能力下降,最终导致靶器官的损害。本研究的主要目的在于：一、探讨加味镇眩方在动物实验研究和临床试验研究中的降压作用,二、探讨加味镇眩方对高血压微血管系统的影响及其他作用机制。第一章加味镇眩方降压作用的实验研究目的加味镇眩方的降压作用。方法将50只SHR大鼠随机分为加味镇眩方高剂量组、加味镇眩方中剂量组、加味镇眩方低剂量组、卡托普利组、对照组,经过12周的干预,分别测定基础血压、心率和2周、4周、6周、8周、10周、12周的血压及心率,以及干预结束后血浆AngⅡ含量。结果血压组间比较显示：各组间血压有统计学差异(F=54.557,P=0.000)；与对照组相比,加味镇眩方低剂量组(P=0.001)、中剂量组(P=0.000)、高剂量组(P=0.000)及卡托普利组(P=0.000)均能降低SHR血压；与高剂量组相比,加味镇眩方低剂量组血压较高(P=0.029)、卡托普利组降压效果更好(P=0.001)均有统计学差异,中剂量组血压与之相比无统计学差异(P=0.249)不同时间的血压水平有统计学差异(F=209.221,P=0.000)。从各个时间点来看,治疗前五组SHR大鼠血压无统计学差异(F=0.850,P=0.501),治疗后2周(F=59.646,P=0.000)、4周(F=56.970,P=0.000)、6周(F=45.251,P=0.000)、8周(F=45.546,P=0.000)、10周(F=57.358,P=0.000)、12周(F=75.150,P=0.000)血压均有统计学差异。加味镇眩方低剂量组(2周P=0.089、4周P=0.002、6周P=0.000、8周P=0.000、10周P=0.000、12周P=0.000)、中剂量组(2周P=0.001、4周P=0.000、6周P=0.000、8周P=0.000、10周P=0.000、12周P=0.000)、高剂量组(2周P=0.000、4周P=0.000、6周P=0.000、8周P=0.000、10周P=0.000、12周P=0.000)及卡托普利组(2周P=0.000、4周P=0.000、6周P=0.000、8周P=0.000、10周P=0.000、12周P=0.000)在治疗后各个时间点血压均比对照组低.加味镇眩方高剂量组在第12周末取得最好的降压效果,与卡托普利组降压效果相当(P=0.020),与低(P=0.000)、中剂量组(P=0.001)相比有统计学差异,其他各个时间点卡托普利组血压均比加味镇眩方高剂量组低(2周P=0.000、4周P=0.000、6周P=0.000、8周P=0.000、10周P=0.000)。在整个实验中,中药组的血压呈平稳持续下降过程,而卡托普利组血压则较快下降,并保持稳定。时间和分组之间有交互效应(F=25.646,P=0.000)组间比较显示：各组间心率有统计学差异(F=11.594,P=0.000)；与对照组相比,加味镇眩方低剂量组(P=0.004)、中剂量组(P=0.000)、高剂量组(P=0.000)均能降低SHR心率,卡托普利组对SHR大鼠心率无明显影响(P=0.297)；与高剂量组相比,加味镇眩方低剂量组(P=0.209)、中剂量组(P=0.596)心率无统计学差异。不同时间的心率水平有统计学差异(F=40.907,P=0.000)。从各个时间点来看,治疗前五组SHR大鼠心率无统计学差异(F=0.981,P=0.427)；治疗后2周(F=3.911,P=0.008)、4周(F=7.220,P=0.000)、6周(F=8.949,P=0.000)、8周(F=9.179,P=0.000)、10周(F=12.621,P=0.000)、12周(F=30.622,P=0.000)心率均有统计学差异,与对照组相比,加味镇眩方低剂量组(2周P=0.045、4周P=0.006、6周P=0.005、8周P=0.000、10周P=0.000、12周P=0.000)、中剂量组(2周P=0.002、4周P=0.001、6周P=0.000、8周P=0.000、10周P=0.000、12周P=0.000)、高剂量组(2周P=0.002、4周P=0.000、6周P=0.000、8周P=0.000、10周P=0.000、12周P=0.000)在治疗后各个时间点均能减慢SHR大鼠心率；卡托普利组在各个时间点与SHR大鼠组心率均无统计学差异。时间和分组之间存在着交互效应(F=7.449,P=0.000)。组间血浆AngⅡ含量的比较：各组血浆AngⅡ含量有统计学差异(F=7.047,P=0.000),与对照组相比,加味镇眩方低剂量组(P=0.015)、中剂量组(P=0.006)、高剂量组(P=0.000)、卡托普利组(P=0.000)均能降低血浆AngⅡ含量；与高剂量相比,加味镇眩方低(P=0.142)、中剂量组(P=0.257)、卡托普利组(P=0.359)在降低血浆AngⅡ方面无统计学差异。结论加味镇眩方可降低SHR大鼠血压,可能与降低SHR大鼠心率及血浆AngⅡ含量有关。第二章加味镇眩方对SHR血管内皮功能的影响目的：通过对自发性高血压大鼠(SHR)内皮依赖的耳廓微血管反应,以及NO的生物合成和生物利用途径的研究,探讨加味镇眩方的降压机制。方法：将50只SHR大鼠随机分为加味镇眩方高剂量组、加味镇眩方中剂量组、加味镇眩方低剂量组、卡托普利组、对照组,12周干预结束,SHR大鼠麻醉后,用激光多普勒系统分别测定耳廓微血管基础血流量以及温热刺激下的耳廓微血管血流量；处死SHR大鼠,分别测定其血浆NO含量、主动脉NOS活性、主动脉cGMP含量。结果：治疗后,各组大鼠耳廓微血管的基础血流灌注量无统计学差异(F=0.633,P=0.641)。将基础血流量作为协变量,采用协方差分析,可以看到,在温热刺激下,各组耳廓微血管血流量有统计学差异(F=55.40,P=0.000)。与对照组相比,加味镇眩方低(P=0.000)、中(P=0.000)、高剂量组(P=0.000)及卡托普利组(P=0.000)均能够增加SHR大鼠耳廓微血管血流量,与其他各组相比,加味镇眩方高剂量均能够增加SHR大鼠耳廓微血管血流量(P=0.000),卡托普利与低剂量中药组血流灌注量无统计学差异(P=0.450)。在温热刺激下,各组之间耳廓微血管血流灌注变化率存在差异(χ2=39.692,P=0.000),对照组对温热刺激的变化最不敏感,血流灌注增加最少,其次为卡托普利组、低剂量组、中剂量组、高剂量组。各组血浆NO(F=3.400,P=0.016)、主动脉NOS活性(F=2.892,P=0.033)及主动脉cGMP含量(F=8.195,P=0.000)有统计学差异。与对照组相比,加味镇眩方低剂量组对SHR大鼠血浆NO(P=0.058)、主动脉NOS活性(P=0.070)无明显影响,加味镇眩方中(P=0.007)、高剂量组(P=0.006)和卡托普利组(P=0.002)均能够明显提高SHR大鼠血浆NO含量,加味镇眩方中(P=0.022)、高剂量组(P=0.009)和卡托普利组(P=0.004)均能够明显提高主动脉NOS活性。与高剂量组相比,加味镇眩方低(P=0.254)、中剂量组(P=0.958)和卡托普利组(P=0.731)血浆NO含量无统计学差异,加味镇眩方低(P=0.393)、中剂量组(P=0.734),卡托普利组(P=0.737)主动脉NOS活性无统计学差异。与对照组相比,加味镇眩方低(P=0.000)、中(P=0.000)、高剂量组(P=0.000)和卡托普利组(P=0.009)均能提高SHR大鼠主动脉cGMP含量；与加味镇眩方高剂量组相比,加味镇眩方低(P=0.306)、中剂量组(P=0.905)主动脉cGMP含量无统计学差异,卡托普利组(P=0.035)主动脉cGMP含量较低。结论加味镇眩方可通过影响SHR大鼠NO/NOS/cGMP合成及利用系统,增加SHR大鼠耳廓微血管血流灌注量,改善血管内皮功能。第三章加味镇眩方对SHR大鼠心肌基质金属蛋白酶-2和左室重塑的影响目的：通过观察加味镇眩方对SHR大鼠左室质量及左室质量指数、左心室心肌胶原浓度、心肌MMP-2蛋白表达,探讨其对左室重塑的作用及其可能机制。方法：将50只SHR大鼠随机分为加味镇眩方高剂量组、加味镇眩方中剂量组、加味镇眩方低剂量组、卡托普利组、对照组,12周的干预结束后,动物称重处死后,电子天平称取左心室和室间隔质量为左心室质量,左室质量指数(LVMD=左心室质量(g)／体重(kg)。称取50mg左心室组织测定羟羟脯氨酸含量,其他组织4℃生理盐水洗净吸干,称重,4%多聚甲醛液中固定后石蜡包埋切片后免疫组织化学方法检测心肌MMP-2蛋白。结果：体重：各组间体重存在差异(F=3.548,P=0.013),与卡托普利组相比,加味镇眩方低(P=0.003)、中(P=0.034)、高剂量组(P=0.004)和对照组(P=0.004)体重增加,与加味镇眩方高剂量组相比,低剂量组(P=0.904)、中剂量组(P=0.401)、对照组(P=1.000)体重无统计学差异。左心室质量：各组间左心室质量存在差异(F=10.868,P=0.000),与对照组相比,加味镇眩方低(P=0.001)、中(P=0.000)、高剂量组(P=0.000)、卡托普利组(P=0.000)左心室质量明显减少。与加味镇眩方高剂量组相比,低剂量组(P=0.013)左心室质量增加,中剂量组(P=0.237)、卡托普利组(P=0.082)左心室质量无统计学差异。左室质量指数：各组间左室质量指数存在差异(F=9.550,P=0.000)；与对照组相比,加味镇眩方低(P=0.001)、中(P=0.000)、高剂量组(P=0.000)、卡托普利组(P=0.001)左心室质量指数明显下降；与加味镇眩方高剂量组相比,低剂量组(P=0.019)、卡托普利组(P=0.017)左心室质量指数增加,中剂量组(P=0.198)左心室质量指数无统计学差异。各组SHR大鼠心肌MMP-2蛋白表达有统计学差异(F=170.854,P=0.000),与对照组相比,加味镇眩方低(P=0.037)、中(P=0.000)、高剂量组(P=0.000)、卡托普利组(P=0.000)均能够降低心肌MMP-2蛋白表达；与高剂量组相比,低剂量组(P=0.000)、中剂量组(P=0.000)、卡托普利组(P=0.000)心肌MMP-2蛋白表达增加。各组大鼠左室心肌胶原浓度有统计学差异(F=25.535,P=0.000),与对照组相比,加味镇眩方低(P=0.000)、中(P=0.000)、高剂量组(P=0.000)、卡托普利组(P=0.000)左室心肌胶原浓度均下降。与加味镇眩方高剂量组相比,低剂量组(P=0.012)、卡托普利组(P=0.004)胶原浓度增加,中剂量组与之无统计学差异(P=0.285)左室质量与终点血压显著相关(r=0.645,P=0.000),左室质量指数与终点血压显著相关(r=0.580,P=0.000)。结论：加味镇眩方能够降低心肌基质金属蛋白酶-2的表达、降低左室质量、降低左室质量指数及左室心肌胶原浓度。第四章加味镇眩方对SHR大鼠心、脑、肾微血管稀疏的影响目的：通过观察加味镇眩方组对SHR大鼠心、脑、肾微血管密度的影响,探讨其对高血压微血管系统的影响及其可能机制。方法：将50只SHR大鼠随机分为加味镇眩方高剂量组、加味镇眩方中剂量组、加味镇眩方低剂量组、卡托普利组、对照组,12周的干预结束后,动物称重处死后,迅速分离左心室,称取50mg左心室组织测定羟羟脯氨酸含量,其他组织4℃生理盐水洗净吸干,称重,立即投入到4%多聚甲醛液中固定12h。同时分离大脑、肾脏,分别称重后,沿冠状面切开,立即投入到4%多聚甲醛液中固定,石蜡包埋后切片,CD34染色免疫组织化学方法检测大鼠心肌、大脑、肾脏微血管密度。结果：各组心肌微血管密度有统计学差异(F=5.212,P=0.002)。与对照组相比,加味镇眩方低(P=0.029)、中(P=0.024)、高剂量组(P=0.000)、卡托普利组(P=0.032)均能够提高SHR大鼠心肌微血管密度。与加味镇眩方高剂量组相比,低剂量组(P=0.026)、中剂量组(P=0.031)、卡托普利组(P=0.023)SHR大鼠脑组织微血管密度无统计学差异各组SHR大鼠心肌微血管密度有统计学差异(F=22.793,P=0.000)。与对照组相比,加味镇眩方低(P=0.000)、中(P=0.000)、高剂量组(P=0.000)、卡托普利组(P=0.007)均能够增加SHR大鼠心肌微血管密度。与加味镇眩方高剂量组相比,低剂量组(P=0.030)、卡托普利组(P=0.000)SHR大鼠心肌微血管密度降低,中剂量组与之无统计学差异(P=0.080)各组肾小管周围微血管密度有统计学差异(F=26.820,P=0.000)。与对照组相比,加味镇眩方低(P=0.000)、中(P=0.000)、高剂量组(P=0.000)、卡托普利组(P=0.000)均能够增加SHR大鼠肾小管周围微血管密度。与加味镇眩方高剂量组相比,低剂量组(P=0.003)、卡托普利组(P=0.001)SHR大鼠肾小管周围微血管密度降低,中剂量组肾小管周围微血管密度与之无统计学差异(P=0.093)。各组肾小球微血管密度有统计学差异(F=29.766,P=0.000)。与对照组相比,加味镇眩方低(P=0.000)、中(P=0.000)、高剂量组(P=0.000)、卡托普利组(P=0.000)均能够提高SHR大鼠肾小球微血管密度。与加味镇眩方高剂量组相比,低剂量组(P=0.000)、卡托普利组(P=0.001)SHR大鼠肾小球微血管密度减少,中剂量组肾小球微血管密度与之无统计学差异(P=0.076)。肾小管周围微血管密度以及肾小球微血管密度与血压的具有相关性,分别为(r=-0.427,P=0.003)和(r=-0.500,P=0.000),心肌(r=-0.172,P=0.247)、大脑(r=-0.255,P=0.084)微血管密度与终点血压无明显相关。结论：加味镇眩方各个剂量组均能改善高血压主要靶器官心、脑、肾脏微血管稀疏的作用,高剂量组在改善脑组织微血管稀疏方面效果最好,中、高剂量组对于心、肾小球及肾小管周围微血管稀疏的效果最好。加味镇眩方对改善微血管稀疏的作用优于西药卡托普利。高血压肾小球及肾小管周围微血管密度与血压具有相关性。第二部分临床部分加味镇眩方治疗原发性高血压的疗效观察目的：通过加味镇眩方对原发性高血压患者血压及中医证侯的疗效观察,探讨其降低血压的临床有效性及安全性。方法：所有病例来源于河南省高血压病医院(河南省中医药研究院附属医院)门诊及高血压病区,性别不限,年龄在18岁-65岁,共90例。试验符合临床试验医学伦理,知情同意后进入临床试验。入组病例须符合高血压低危或中危(1～2级)诊断及中医肝火亢盛证或痰湿壅盛证诊断。90例患者随机分为中药治疗组和卡托普利对照组,中药治疗组45例,卡托普利对照组45例,中药组给予加味镇眩方治疗；对照组以西药卡托普利片(12.5mg,日二次),疗程4周。分别观察治疗前后的血压水平、中医证候积分及安全性指标。两组病例在性别(χ2=3.455,P=0.063)、年龄(t=0.429,P=0.669)、病程(t=—1.157,P=0.251)无统计学差异。两组治疗前收缩压无统计学差异(Z=-0.543,P=0.587)；舒张压无统计学差异(Z=-1.091,P=0.275)；中医证侯积分,无统计学差异(Z=-0.734,P=0.463)。结果：不同时间收缩压有统计学差异(F=369.678,P=0.000),治疗后两组收缩压均下降；两组之间收缩压变化无统计学差异(F=0.132,P=0.719),时间和分组之间无交互效应(F=1.120,P=0.297)。不同时间舒张压有统计学差异(F=50.230,P=0.000),治疗后两组舒张压均下降,两组之间舒张压变化无统计学差异(F=4.010,P=0.053)。时间和分组之间无交互效应(F=0.005,P=0.944)。不同时间中医证侯积分有统计学差异(F=368.841,P=0.000),治疗后两组中医证侯积分均降低,两组之间中医证侯积分变化有统计学差异(F=10.462,P=0.003),治疗组优于对照组。时间和分组之间有交互效应(F=59.588,P=0.000)。两组在降压疗效方面无统计学差异(Z=-0.759,P=-0.448)。两组在改善中医证侯方面有统计学差异(Z=-5.147,P=0.000),治疗组优于对照组。安全性评价：两组患者治疗前后实验室各指标检测均正常,卡托普利组因皮疹剔除1例,因咳嗽脱落6例,加味镇眩方组因无法坚持服药脱落7例。结论：加味镇眩方和卡托普利(25mg/d)均能降低高血压患者的血压,两者在降压方面疗效相当,收缩与舒张压下降均无显著差异。中药与卡托普利均能够改善中医症候体征,中药组疗效显著优于卡托普利组。更多还原

【Abstract】 In China, with the improvement of living standards and lifestyle changes, the incidence of hypertension increased steadily, the 2002 National Nutrition and Health Survey data show that adult prevalence rate was 18.8%, there are about 160 million patients with hypertension. As the most common chronic disease, hypertension is one of most important basis disease led to stroke, coronary heart disease and renal failure and other diseases.Although Western medicine could lower blood pressure quickly, efficacy and the advantages of good compliance, but the side effects of antihypertensive drugs was more frequent, many patients had to give up treatment. Chinese medicine treatment of hypertension has unique advantages, such as blood pressure stable and durable, while alleviating symptoms, less side effects. Professor Chen Baotian according to the "dizzy all the Feng, all belonging to the liver," "do not dizzy without Tan " and related theories of Chinese medicine, combined with decades of clinical experience, create Modified ZhenXuanFang for the treatment of hypertension, and had a good effect. If rational development and wide clinical application, it will help complement the disadvantage of western medicine in decreasing blood pressure stablely and alleviating symptoms in patients with hypertension, reducing blood pressure medication side effects, improve health quality of nearly 200 million people hypertensive patients in chinathe pathophysiology of microcirculation has paly a more important role in hypertension. Reduction in microvessel density, not only led to decreased blood parallel pathway, and lead to increased peripheral vascular resistance, thereby promoting the continued increase in arterial blood pressure, reduced microvessel density, but also enables organizations to the level of physical energy metabolism, storage capacity and material exchange capacity decreased, leading to target organ damage at the same time. The main purpose of this study:first, to explore effect of Modified ZhenXuanFang on blood pressure in experimental and clinical research,senong, to explore effects of Modified ZhenXuanFang on microvascular system and other mechanism.ChapterⅠThe study of the effect of Modified ZhenXuanFang on blood pressure of spontaneously hypertensive ratsObjective To explore the effect of Modified ZhenXuanFang on blood pressure of spontaneously hypertensive ratsMethods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose Modified ZhenXuanFang group,the captopril group and SHR control group, Blood pressure and heart rate were recorded from the tail at the beginning and every 2 weeks during 12 weeks treatment. At the end of the experiment, animals were sacrificed.the plasma AngⅡconcentration were measured.Rusults Test of between-subjects effect in decreasing blood pressure indicate: significant differences were seen in the differences groups of SHR (F=54.557,P= 0.000),When compared with control group, the low-dose groups (P=0.001),the middle-dose (P=0.000) and high-dose (P=0.000) Modified ZhenXuanFang group,the captopril group (P=0.000) all show significant differences in decreasing blood pressure of spontaneously hypertensive rats.When compared with high-dose Modified ZhenXuanFang group, the low-dose groups show significantly higher blood pressure (P=0.029), the captopril group show significantly lower blood pressure (P =0.001),and no significant differences were seen in middle-dose group (P= 0.249).Significant differences were seen in differences time (F= 209.221,P=0.000).There were no significant differences in blood pressure at the beginning (F=0.850,P=0.501) and significant differences were seen in blood pressure in differences group on second week (F=59.646,P=0.000),fourth week (F=56.970,P=0.000),sixth week (F=45.251,P=0.000),eighth week (F= 45.546,P=0.000),tenth week (F=57.358,P=0.000),twelfth week (F= 75.150,P=0.000).When compared with control group, the low-dose (second week P =0.089,fifth week P=0.002,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),the middle-dose (second week P=0.001,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),high-dose (second week P=0.000,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000) Modified ZhenXuanFang group and the captopril group (second week P=0.000,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000) all show significant in lowering blood pressure.The high-dose group show the best effect in reduction in blood pressure and show significant differences compared with the low-dose (P=0.000) and middle-dose groups (P=0.001) and no significant differences compared with captoril group (P=0.020)in decreasing blood pressure in 12 weeks. the captopril group were more significant in reduction in blood pressure at other times (second week P= 0.000,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000).Ttraditional Chinese medicine group could steady decrease blood pressure in the experiment,and captoril group could quickly decrease blood pressure. Time and group show significant crossover effect (F=25.646,P=0.000)Test of between-subjects effect in decreasing blood pressure indicate:significant differences were seen in the differences groups of SHR(F=11.594,P=0.000); When compared with control group, the low-dose groups (P=0.004),the middle-dose (P =0.000) and high-dose (P=0.000) Modified ZhenXuanFang group all show significant differences in decreasing heart rate of spontaneously hypertensive rats. no significant differences was seen in captoril group(P=0.297). When compared with high-dose Modified ZhenXuanFang group, the low-dose group (P=0.209) and middle-dose group (P=0.596) show no significant differences in heart rate of spontaneously hypertensive rats.Significant differences were seen in differenent time (F=40.907,P=0.000).There were no significant differences in heart rate at the beginning (F= 0.981,P=0.427) and significant differences were seen in heart rate in differences group on second week (F=3.911,P=0.008).fourth week (F=7.220,P=0.000),sixth week (F=8.949,P=0.000),eighth week (F=9.179,P=0.000),tenth week (F= 12.621,P=0.000),twelfth week (F=30.622,P=0.000).When compared with control group, the low-dose (second week P=0.045,fifth week P=0.006,sixth week P=0.005,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),the middle-dose (second week P=0.002,fifth week P=0.001,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),high-dose (second week P =0.002,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000) Modified ZhenXuanFang group all show significant in decreasing heart rate. the captopril group show no significant differences (second week P=0.068,fifth week P=0.343,sixth week P=0.448,eighth week P=0.210,ten week P=0.416,twelfth week P=0.358) at any time in the experiment.Time and group show significant crossover effect (F=7.449,P=0.000) Test of between-subjects effect in plasma AngⅡconcentration indicate: significant differences were seen in differenent in different group (F= 7.047,P=0.000), When compared with control group, the low-dose (P=0.015), middle-dose (P=0.006), high-dose (P=0.000) Modified ZhenXuanFang group and captopril group (P=0.000) all show significant differences in plasma AngⅡconcentration of spontaneously hypertensive rats. When compared with high-dose Modified ZhenXuanFang group, the low-dose group (P=0.142),the middle-dose (P=0.257) and the captopril group (P=0.359) show no significant differences in decreasing plasma AngⅡconcentration of spontaneously hypertensive ratsConclusion Modified zhenxuan granules could decrease blood pressure of spontaneously hypertensive rats, perhaps by decreasing heart rate and plasma AngⅡconcentration.ChapterⅡThe study of the effect of Modified ZhenXuanFang on endothelial function in spontaneously hypertensive ratsObjective To explore the effect of Modified ZhenXuanFang on blood pressure of spontaneously hypertensive rats By measuring noninvasively endothelial function in the auricular skin microcirculation by monitoring with a laser-Doppler system the blood flow changes induced by warming of the skin and biosynthesis and bioavailability of NO.Methods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose groups Modified zhenxuan fang group,the captopril group and SHR control group.At the end of the experiment, animals were anesthetized and measured auricular skin blood flow changes by Laser Doppler system before and after local warming.Then animals were sacrificed. The plasma NO concentration,arteriae aorta nitric oxide synthase (NOS) activity and arteriae aorta cyclic guanylic acid (cGMP) concentration were measured. Rusults The baseline auricular skin blood flow perfusion units show no Significant differences in the differences groups of SHR (F=0.633,P=0.641).The basis of blood flow as a covariate, using analysis of covariance, we find auricular skin blood flow perfusion units were significantly different in each group (F=55.40,P=0.000). Compared with the control group, the low-dose (P=0.000),the middle-dose (P=0.000), high-dose group (P=0.000) Modified zhenxuan fang group and the captopril group (P=0.000) could improve auricular skin blood flow perfusion units of SHR. high-dose group could improve auricular skin blood flow perfusion units compared with other groups (P=0.000).captopril and low dose group show no significant difference in auricular skin blood perfusion units (P= 0.450).the rate of change of the auricular microvascular perfusion units differences (χ2=39.692, P=0.000) before and after local warming, the control group increased blood flow perfusion units at least, followed by captopril group, low dose group, middle dose group and high dose group.The plasma NO (F=3.400, P=0.016), aortic NOS activity (F=2.892, P= 0.033) and aortic cGMP content (F=8.195, P=0.000) were significantly different.Compared with the control group, Modified ZhenXuanFang low dose group SHR plasma NO (P=0.058), aortic NOS activity (P=0.070) had no significant difference, Modified ZhenXuanFang middle dose group (P=0.007), high dose group (P=0.006) and captopril group (P=0.002) were able to significantly increase the plasma NO concentration in SHR,modified ZhenXuanFang middle dose (P=0.022), high dose group (P=0.009) and captopril group (P=0.004) were able to significantly increase aortic NOS activity. Compared with the high dose group, modified Zhenxuan low-dose (P=0.254), middle dose group (P=0.958) and captopril group (P=0.731) showed no significant difference in plasma NO, modified Zhenxuan low-dose group (P=0.393), middle dose group (P=0.734), captopril group (P= 0.737) show no significant difference in aortic NOS activity.Compared with the control group, modified Zhenxuan low-dose group (P= 0.000), middle dose group(P=0.000), high dose group (P=0.000) and captopril group (P=0.009) could increase aortic cGMP concentration; Compared with Modified ZhenXuanFang high dose group, low dose group (P=0.306), middle dose group (P=0.905) show no significant differences in aortic cGMP, the captopril group was lower(P=0.035) in Aortic cGMP concentration,Conclusion Modified ZhenXuanFang could affecte NO/NOS/cGMP biosynthesis and bioavailability of SHR, increase the SHR rat auricular microvascular perfusion to improve endothelial function.ChapterⅢThe study of modified ZhenXuanFang on SHR myocardial matrix metalloproteinase-2 and left ventricular remodelingObjective:To explore the effect of Modified ZhenXuanFang on left ventricular remodeling and possible mechanisms by observing spontaneously hypertensive rats left ventricular mass, left ventricular mass index, left ventricular collagen concentration, myocardial expression of MMP-2 after treatment.Methods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose Modified ZhenXuanFang group,the captopril group and SHR control group, At the end of the experiment, animals were sacrificed. electronic balance to take the quality of left ventricular and ventricular septal left ventricular mass, left ventricular mass index (LVMD=left ventricular mass (g)/ body weight (kg). Weighed 50mg of left ventricular tissue to detect hydroxyproline, other left ventricular tissue was washed in 4℃saline,dried, weighed, and 4% paraformaldehyde solution fixed and paraffin embedded and detected myocardial MMP-2 protein by immunohistochemistry.Results:Weight:animals’weight differ among groups (F=3.548, P=0.013), compared with captopril group, Modified ZhenXuanFang low dose group (P= 0.003),middle dose group (P=0.034), high dose group (P=0.004) and control group (P=0.004) weight heavier, compared with Modified ZhenXuanFang high dose group, low dose group (P=0.904), middle dose group (P=0.401), control group (P=1.000) weight show no significant difference.Left ventricular mass:Left ventricular mass differ among the groups (F= 10.868, P=0.000); compared with the control group, modified ZhenXuanFang low dose group (P=0.001), middle dose group (P=0.000), high dose group (P=0.000), captopril group (P=0.000) decreased left ventricular mass significantly; Compared with modified ZhenXuanFang high dose group, low dose group (P=0.013) left ventricular mass increased, the middle dose group (P=0.237), captopril group (P= 0.082) left ventricular mass show no signifcant differences.Left ventricular mass index:the group left ventricular mass index differ among the groups (F=9.550, P=0.000); compared with the control group, modified ZhenXuanFang low dose group (P=0.001), middle dose group (P=0.000), high dose group (P=0.000), captopril group (P=0.001) significantly reduced left ventricular mass index; compared with modified ZhenXuanFang high dose group, low dose group (P=0.019), captopril group (P=0.017) left ventricular mass index increase,middle dose group (P=0.198) show no significant difference in left ventricular mass index.Myocardial expression of MMP-2 protein were significantly different among the groups (F=170.854, P=0.000), compared with the control group, modified ZhenXuanFang low dose group (P=0.037), middle group (P=0.000), high dose group (P=0.000), captopril group (P=0.000) were able to reduce the myocardial expression of MMP-2 protein; and compared with the high dose group, low dose group (P=0.000), middle dose group (P=0.000), captopril group (P=0.000) myocardial MMP-2 protein expression were up regulation.Left ventricular myocardial collagen concentration were significantly different (F=25.535, P=0.000), compared with the control group, modified ZhenXuanFang low dose group (P=0.000), middle dose group(P=0.000), high dose group (P= 0.000), captopril group (P=0.000) left ventricular collagen concentration decreased. Compared with modified ZhenXuanFang low dose group, low dose group (P= 0.012), captopril group (P=0.004) increase in the concentration of collagen, the middle dose group show no significant difference (P=0.285)Left ventricular mass was significantly correlated with end blood pressure (r= 0.645, P=0.000),left ventricular mass index was significantly correlated with end blood pressure (r=0.580, P=0.000).Conclusion:Modified ZhenXuanFang could reduce the expression of matrix metalloproteinase-2, lower left ventricular mass, reduced left ventricular mass index and left ventricular collagen concentrationChapterⅣThe study of modified ZhenXuanFang on microvascular rarefaction in heart,brain and kidney of SHRObjective To explore Modified ZhenXuanFang on microvessel density in hypertension and its possible mechanism by observing the Modified ZhenXuanFang group, captopril group, spontaneously hypertensive rats control group myocardial, cerebral and renal microvessel density.Methods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose Modified ZhenXuanFang group,the captopril group and SHR control group, At the end of the experiment, animals were sacrificed, left ventricular tissue was washed in 4℃saline,dried, weighed, and 4% paraformaldehyde solution fixed and paraffin embedded, At the same time separating the brain, kidney, respectively, after weighing, cut along the coronal plane, and immediately put into 4% paraformaldehyde solution fixed and paraffin-embedded sections,CD34 immunohistochemical staining to detect myocardial, cerebral, renal MVDResults Myocardial microvessel density in each group were significantly different (F=5.212, P=0.002). Compared with the control group, the low dose fang group (P=0.029), middle dose group (P=0.024), high dose Modified zhenxuan group (P=0.000), captopril group (P=0.032) were able to improve the SHR cerebral microvessel density. Compared with high-dose Modified ZhenXuanFang group,low dose group (P=0.026), middle dose group (P=0.031), captopril group (P=0.023) SHR cerebral microvessel density show no significant differenceCerebral microvessel density in each group were significantly different (F=22.793,P=0.000). Compared with the control group, the low dose group (P= 0.000), middle dose group(P=0.000), high dose Modified ZhenXuanFang group (P= 0.000), captopril group (P=0.007) were able to improve the cerebral microvessel density. Compared with high-dose Modified ZhenXuanFang group, low dose group (P=0.030), captopril group (P=0.000) cerebral microvessel density decreased,the middle dose group show no significant difference (P=0.080).Peritubular microvessel density in each group were significantly different (F= 26.820, P=0.000). Compared with the control group, the low dose group (P=0.000), middle dose group (P=0.000),high dose group (P=0.000), captopril group (P= 0.000) were able to improve the SHR Peritubular microvessel density. Compared with high-dose Modified ZhenXuanFang group, low dose group (P=0.003), captopril group (P=0.001) Peritubular microvessel density decreased, the middle dose group show no significant difference (P=0.093).Glomerular microvessel density in each group were significantly different (F =29.766, P=0.000). Compared with the control group, the low dose group (P= 0.000), middle dose group (P=0.000),high dose group (P=0.000), captopril group (P=0.000) were able to improve the SHR glomerular microvessel density. Compared with high-dose Modified ZhenXuanFang group, low dose group (P=0.003), captopril group (P=0.001) SHR glomerular microvessel density decreased, the middle dose group show no significant difference (P=0.076).Peritubular and glomerular microvessel density all correlated with end blood pressure, respectively (r=-0.427, P=0.003) and (r=-0.500, P=0.000), myocardial (r=-0.172, P=0.247), cerebral tissue (r=-0.255, P=0.084) microvessel density was not related with end blood pressure.Conclusion All dose Modified ZhenXuanFang group can improve the main target organs cardiac, cerebral, renal microvessel density, high-dose group improve cerebral microvascular best, middle and high dose group improve myocardial, peritubular, glomerular microvessel density best. Modified ZhenXuanFang was superior to captopril in improving microvessel density. glomerular and peritubular microvessel density correlated with end blood pressure.Clinical part The study of the effect of Modified ZhenXuanFang on essential hypertensionObjective To explore the clinical effectiveness of lowering blood pressure and safety of Modified ZhenXuanFang by study the effect of Modified ZhenXuanFang on essential hypertension and TCM Symptom scores.Methods All patients from the Henan hypertension hospital (Henan Academy of Traditional Chinese Medicine Hospital) out-patient department and in-patient department, either sex, aged 18-65 years old, a total of 90 cases. Test line with clinical trials of medical ethics, all patients enter clinical trials after informed consent.All the hypertensive patients should be consistent with low-risk or middle-risk group(1-2) diagnosis and ganhuokangsheng or tanshiyongshen of Chinese medicine diagnosis.90 cases were randomly divided into treatment group and captopril group,45 cases of Chinese medicine treatment group and 45 cases of captopril group, treatment for 4 weeks. Medicines for the Treatment of Modified ZhenXuanFang, the control group for captopril tablets (12.5mg,.b.i.d) for 4 weeks. The blood pressure, the symptom score and the safety index were measured before and after treatment. the gender (χ2=3.455,P=0.063), age (t=0.429, P=0.669)and Course of disease (t=-1.157, P=0.251), systolic blood pressure (Z=-0.543, P= 0.587), diastolic blood pressure (Z=-1.091, P=0.275),TCM syndrome score (Z=-0.734, P=0.463)show no significant difference in two groups before treatment.Results Systolic blood pressure were significantly different at different times (F=369.678, P=0.000), systolic blood pressure change between the two groups was not significantly different (F=0.132, P=0.719), Time and group show no significant crossover effect (F=1.120, P=0.297). Diastolic blood pressure were significantly different at different times (F=50.230, P=0.000), diastolic blood pressure change between the two groups was not significantly different (F=4.010, P=0.053). Time and group show no significant crossover effect (F=0.005, P=0.944).TCM syndrome score in different time points were significantly different (F= 368.841, P=0.000), TCM syndrome score between the two groups were significantly different (F=10.462,P=0.003). Time and group show significant crossover effect (F =59.588, P=0.000). Antihypertensive effect in two groups show no significant difference (Z=-0.759, P=-0.448). Treatment group was superior to the control group in relieving syndromes (Z=-5.147, P=0.000). Safety evaluation:two patients laboratory indexes were safe before and after trail, the captopril group removed 1 case because of erythra,6 cases of loss due to cough,7 cases were removed because of stopping to take chinese medicine in Modified ZhenXuanFang.Conclusion Modified ZhenXuanFang and captopril (12.5mg/d) could lower blood pressure, the two groups showed no significant difference in lower systolic and diastolic blood pressure. Two medicine could relieving syndromes both, modified ZhenXuanFang was superior to captopril in relieving syndromes更多还原