【作者】 梁启军；

【导师】 王鹏；

【作者基本信息】 湖北中医药大学 ， 中医内科学， 2010， 博士

【摘要】 一实验设计和检测指标选择的指导理论：三维宏观态势理论1三维宏观态势理论任何生命体只有物质、能量、时空三个生命维度；生命过程就是生命的自利、自洁和自稳的维持过程。以物质、能量、时空三个维度体现的人体生命首先是一个物质、能量摄入、贮存、利用和排出的动态过程。对于不利于自体的物质,包括多量的人体正常需要物质、有害的理化物质、细菌和病毒等致病微生物、过多积聚的自体生理产物等,人体在长期化中已经形成了一套完善的自稳免疫机制,在宏观上表现为祛其外出、自洁人体的宏观免疫态势。宏观上人体正气祛邪外出的路径选择或者最小化邪气的损害有以下五个基本态势：一是从哪里进来就从哪里出去,因为进处即是易出处。二是从最近或最便利的人体大型外通腔道祛出。三是通过到达体表最近路径祛出,比如形成瘘道。四是通过微观路径趋出。五是就地限制包裹,但祛邪外出总是人体宏观免疫的最终导向。在疾病的发生、发展和转归过程中,人体的宏观免疫态势就会表现出来；同时人体正气和邪气各自情况也就通过物质维(以正气虚实和邪气盛衰为表现)、能量维(以体温和热性为表现)、空间维(以病位为表现)人体生命三维参量变化表现出来,因此正邪斗争总的态势就呈现出来。中医是从人体固有的生命三维、用宏观态势描述法客观地表述人体生理病理规律和现象,运用二分法描述人体正气与邪气,把人体所有自利己体的物质功能作为一个整体,称作正气,而所有致病因素作为另一个整体,称作邪气,正气顺应上述宏观免疫态势对邪气形成祛之离体的总斗争态势,然后根据此态势确定总的治疗原则——三维纠偏、因势利导、扶正祛邪。因此中医是三维宏观态势医学。与中医理论相对映,传统中药学描述的是三维一势：1)药性——能量维,2)五味——物质维(成分),3)归经(特定组织、器官即空间选择性作用)——空间维,4)升、降、沉、浮(四气)——运动方向的宏观态势。中药复方的总配伍原则也是三维纠偏、因势利导、扶正祛邪。2三维宏观态势理论指导下的中药复方实验指标选择思路2.1体现生命物质、能量、时空三维,即体现中医八纲辨证的虚实、寒热、表里之纲：物质维指标——一切关于宏观和微观具体生命物质的指标,都属于物质维。能量维指标——体温、ATP、ATP酶、主观的冷热感觉等指标。时空维指标——与时间、季节、空间、环境、体积、大小、部位等相关指标。2.2体现生命整体或局部运动态势：趋向多动、多语,或趋向沉静等体现整体运动态势的指标。趋向多饮多食、血凝气滞等体现局部运动态势的指标。2.3体现扶正祛邪的中医基本治疗思维：扶正指标——体现组织修复、功能趋向完整或提高的指标。祛邪指标——体现内、外邪气变化的指标。体现正邪斗争态势和祛邪离体路径的指标——前者含在扶正、祛邪指标之内,后者如汗出、陈痰咳出、瘀血排除、积便排出等。二、益肺活血汤治疗肺心病初期家兔模型机理实验研究目的：在三维宏观态势理论指导下,选择检测指标,通过对益肺活血汤治疗肺心病初期家兔模型的实验研究,初步揭示其治疗机理,为益肺活血汤进一步研究提供基础,也为研究中医、中药、复方提供一个理论思路。方法：购进北京沙河通利试验养殖场普级家兔大耳白40只,许可证批号：scxk北京2005-0003,雌雄不拘,每只2kg左右,随机分为正常对照组、肺心病模型组、益肺活血汤治疗组、卡托普利治疗组,每组10只。正常对照组不造模,肺心病模型组、益肺活血汤治疗组、卡托普利治疗组一并造模。参考浙江医科大学俞秋棠家兔肺心病模型建立方法,从兔耳缘静脉注射0.7%三氯化铁溶液,隔日1次,复制肺心病模型；从行为学、二维超声心动图判断肺心病模型已造成,停止造模。造模结束后,每只家兔从耳缘静脉取血、观察血液宏观形态,然后按需留取血浆或血清,检测指标AngⅡ、ET、TXA2/PGI2、NO、TNF-α取血后,正常对照组和肺心病模型组依前法喂养,以备用的益肺活血汤膏粘剂喂药益肺活血汤治疗组,以卡托普利喂药卡托普利治疗组,每天均喂药,喂药20天。因实验中途每组家兔均死亡两只,给药结束时每组均余兔8只,共32只。首先观察各组家兔行为学特征,随后耳缘静脉取血,观察血液宏观形态、提取血浆、血清,做造模结束时相同指标检测,然后杀死家兔观察心、肺宏观形态学特征,并取材家兔肺组织,做心、肺切片、检测肺组织细胞增殖和凋亡现象及ATPase活性变化。结果：第一部分：从各组的心、肺宏观形态、心、肺颜色和色泽、赘生物有无、心肌与浆膜关系改变等方面的差异,显示益肺活血汤疗效良好。心、肺切片从心肌变化、肺小动脉变化、血栓改变、炎性细胞行为等微观形态学方面也证明益肺活血汤的疗效。第二部分：血管活性因子AngⅡ、ET、TXA2/PGI2、NO及TNF-α检测结果,用方差分析、q检验分析如下：益肺活血汤可以显著提高血清NO含量(p<0.01)；有降低血液中AngⅡ、ET含量及TXA2/PGI2的比例的趋势(p>0.05)；对TNF-α无影响(p>0.05),且治疗后各组间TNF-α无差异(p>0.05)第三部分：益肺活血汤有明显促进肺组织细胞增殖作用(p<0.05),各组均未观察到凋亡现象。第四部分：益肺活血汤有明显提高肺组织Na+-K+-ATPase活性作用(p<0.01),对Ca2+-Mg2+-ATPase活性无明显影响(p>0.05)。结果分析：肺心病虽有诸多证型,但肺气(阳)虚兼血瘀是主要证型；益气(阳)活血是肺心病中医治疗的主要方法和普遍思维之一。益肺活血汤是中医大家董建华院士治疗慢性肺心病的临床验方,是在补中益气汤和血府逐瘀汤基础上加减而成,方中黄芪、党参、白术、甘草补益脾肺之气,柴胡、枳壳、桔梗、陈皮理气,川芎、牛膝、当归、桃仁、红花、赤芍活血化瘀,生地黄养阴生津。全方单从药性上看,黄芪微温,白术、枳壳、陈皮、川芎、当归、红花温,党参、桔梗、甘草、桃仁、牛膝平,赤芍、柴胡微寒,生地黄寒；微温、温、平、微寒、寒药量的比例是10：42：40：11：9,全方温性突出,实是温阳为主的益气温阳活血化瘀之方。在形态学方面,心、肺宏观形态学特征、心、肺切片、血液宏观形态的差异就直观证明益肺活血汤有疗效。益肺活血汤治疗组肺组织细胞增殖现象明显强于其它组,既是补益作用的体现,又是补益作用的佐证。Na+-K-+ATPase活性的实验结果表明提高Na+-K-+ATPase活性是以益气温阳活血化瘀之法立方的益肺活血汤治疗肺心病的机制之一。实验证明益肺活血汤能明确提高血液中NO的含量,仅有降低血液中AngⅡ、ET含量及TXA2/PGI2的比例的趋势,所以对血管活性物质影响方面,有待进一步研究。益肺活血汤对TNF-α无影响,表明抗炎或祛邪不是其主要作用,治疗后各组间TNF-α无差异表明益肺活血汤是安全的,无致瘤作用。结论：此次实验从组织细胞修复角度(促进肺组织细胞增殖)、能量代谢角度(增强肺组织Na+-K+-ATPase活性)、血管活性物质角度(增加血管舒张因子NO的含量等)、抗炎及副作用角度(治疗前后TNF-α的变化及治疗后TNF-α的含量无统计学差异)初步揭示了益肺活血汤,即益气温阳活血化瘀法对肺心病的治疗机理,并初步否定了最不希望出现的副作用——致瘤作用。实验指标选择也充分体现益肺活血汤扶正(促进肺组织细胞增殖、增强肺组织Na-+K+-ATPase活性)、纠正病理态势(调节血管舒张和收缩物质的、消散血栓、降低血液粘稠度等)等方面疗效,证明益肺活血汤的疗效是客观的,值得进一步研究。更多还原

【Abstract】 I The guiding theory of designing experiment and selecting test indicators:three-dimension-macro-trend theory1Three-dimension-macro-trend theoryAny life only has the life dimensions of matter, energy, space-time, and the life is a course of self-serving,self-cleaning and self-smoothening. With the three dimensions of matter,energy, and space-time, the human life is the dynamic process of material and energy intake, storage, use and discharge first and foremost. The body has formed a set of improved immune mechanism in the long run of evolution which shows the macro-immune-trend (or law) in the macro perspective to drive out the substances that are not conducive to the body and that include harmful physical and chemical substances,bacteria,viruses and other pathogenic micro organisms, excessive normal life needs or accumulation of physical products from the body. Body has the following five basic macro trends to drive out the pathogenic factors or minimize the damages.First, where out is just the one in because the place coming from is where easy to go out.Second, the pathogenic factors are drived out from the nearest or the most convenient large body cavity linking the outside. Third, they are drived out through fistula developed by the nearest path to the surface. Fourth, the paths are the micro ones. Fifth, they are beleaguered where they develop. But driving-out is always ultimate of the human macro-immune-trend.The body’s macro-immune trend show in the disease’s occurrence, development and transfer process,and in the same time, the changes of healthy minarals and energy and the pathogenic factors are also demonstrated through matter dimension (by strongth or weakness of healthy minarals and energy and the pathogenic factors),the energy dimension(by temperature and the heat or cold feeling),spatial dimension (by disease location).so the overall fighting trend between them appears.From the inherent three life dimensions of human body, Chinese Medicine describs the macro trend of human physiological and pathological laws and phenomena, and describs all healthy minerals and functions as Zhengqi and the pathogenic factors as Xieqi by dichotomy. The general therapy principle is set out based on struggling trend between them that is three-dimension correction, strenthening Zhengqi and driving out pathogenic factors according to the trend of Zhengqi frighting Xieqi.Traditional Chinese Medicine is three-dimension-macro-trend medicine which cures diseases according to the trend of Zhengqi frighting Xieqi and by correctting abnormalities of life three dimensions. Traditional Chinese Medicine also describes the three dimensions and one trend of Chinese medicine:1)The heat or cold of the medicinethe energy dimension,2)five taste-matter dimension(components),3) the target meridians and collaterals of Chinese medicine (specific tissues,organs,namely, the space selective role)theory-space dimension,4) lift ups and downs(4 gases)--the macro-trend of movement direction. The general principle of Chinese herbal compound is three-dimension correction, strenthening healthy minerals and energy and driving out pathogenic factors according to the trend of Zhengqi frighting Xieqi,too.2 Selecting test indicators2.1 To reflect the life three dimensions of material,energy, space-time,namely, reflect the differentiation of eight principle syndromes of Traditional Chinese Medicine.Matter dimension-all macro-and micro-indicators of specific life substances belong to the matter dimension.Energy-dimension-body temperature, ATP, ATP enzyme, the subjective hot and cold feeling,and so on.Space-time dimension-time, season, space, environment, volume, size, location and other relevant indicators and so on.2.2 To reflect the the whole or partial life moving trendThe indicators of hyperactivity, multi-language, or silence to reflect the the whole moving trend;The indicators of polydipsia,blood clotting Qi stagnation to reflect the local moving trend.2.3 To reflect the Chinese Medicine thinking to strengthen the healthy minerals and energy and driving out pathogenic factors:Strengthening healthy minerals and energy-the indicators to reflect tissues repair,functions completed or improvedDriving out pathogenic factors-the indicators to reflect the changes of internal and external pathogenic factors.Showing the trend btween the healthy minerals and energy and the pathogenic factors,and the path of pathogenic factors being drived out.The former indicators can be included in the ones of strengthening healthy minerals and energy or driving out pathogenic factors.The latter can include a lot,such as sweating, coughing out old phlegm, expeling out congestion, discharging the dry plot, etc.II Experimental researchObjective:The therapeutic mechanisms were initially reveal that the rabbit models of cor pulmonale were treated with Decoction of Replenishing Lungs and Activating Blood Circulation by the experiment under the guide of Three-dimension-macro-trend theory. The results and conclusions will become the basis to further research the Decoction and provide a thinking for sdudy of Traditional Chinese Medicine and it’s medicine compound.Methods:40 average class white rabbits with big ears,purchased from the Test Farm of Shahe in Beijing,permits Lot:scxk Beijing 2005-0003,no differentiation of the male and female, about 2kg each, and they divided randomly into normal control group, model group of cor pulmonale,treatment group of Decoction of Replenishing Lungs and Activating Blood Circulation, and the captopril treatment group,10 rabbits in each group, The control group was not modeled, and the other groups were modeled altogether,and all groups were fed with the same feed.Referring to rabbit model of cor pulmonale made by Yu Qiu-tong in Zhejiang Medical University, the models were copied by injection of 0.7% ferric chloride solution from rabbit ear vein one time every other day. The modeling times were 20. From behaviors and two-dimensional echocardiography, cor pulmonale model had successed, modeling ended. After modeling,each rabbit was taken blood from the ear vein for Observation of blood macro-morphology and detection of AngⅡ,ET, TXA2/PGI2, NO, TNF-α. firstly, Control and Model group were fed only after the former method then, and China group fed with the viscose cream, West group fed with captopril,and two medicines were all given once a day. Eight rabbits survived to the end of delivery in each group, and the total was 32. Firstly, to observe and not the behavior characteristics of rabbits,and to take blood from ear vein for the observation of blood macro-morphology and extraction of plasma or serum, for the same detection indicators at the end of modeling, then to kill the rabbits for observation of general hear tand lung morphological characteristics and derive rabbit lung tissues to make the heart and lung biopsy, detection of lung cell proliferation and apoptosis and the changes in ATP enzyme activity.Results:PartⅠ:From the difference of general morphology, the color,xcrescences of heart and lungs,and the changes between heart and it’s pericardium, a good efficacy has shown in Decoction of Replenishing Lungs and Activating Blood. The efficacy also was proved by the micro-morphological changes in cardiac muscle, pulmonary artery, thrombosis and behavors of inflammatory cellsPartⅡ:By analysis of variance and q test of vasoactive factors AngⅡ,ET, TXA2/PGI2,NO, and inflammatory cytokine TNF-α,conclusions are as follows:Decoction of Replenishing Lungs and Activating Blood Circulation can very significantly improve the serum NO levels (p<0.01);and has the trend to lower blood Ang II,ET and TXA2/PGI2, and has no effect to TNF-α,and no significant difference on TNF-αappear between th ebefore and after treatment and among the four groups after treatment.PartⅢ:Decoction of Replenishing Lungs and Activating Blood Circulation has significant effect in promoting cell proliferation in lung tissue (p<0.05),and apoptosis is not observed in each group.PartⅣ:Decoction of Replenishing Lung and Activating Blood Circulation can improve markedly Na+-K+-ATP enzyme activity of lung tissue (p<0.01),and has no significant effect to Ca2+-Mg2+-ATP enzyme activity.Analysis of results:Despite cor pulmonale can show many syndromes,the deficiency of lung-Qi(and Yang)and blood stasis of lung are the main and common pathophysiological features;and replenishing Qi (and Yang) of lung and activating blood circulation are a general thinking and main methods for Traditional Chinese Medicine to treat cor pulmonale. Decoction of Replenishing Lungs and Activating Blood Circulation was a clinical prescription of Traditional Chinese Medicine created by Academy Dong jian-hua for the people with chronic cor pulmonale which was created on the basis of Decoction for Reinforcing Middle-warmer and Replenishing Qi and Decoction for Dissipating Blood Stasis in Chest,and in which Astragalus,Lu Codonopsis,White Atractylodes Rhizome and Licorice can reinforce Qi of spleen and lungs;Bupleurum, the Peel of Bitter Orange,Balloonflower Boot and Dried Old Orange Peelcan can regulate the flow of Qi contribute the ascending trend of lungs and spleen; Chuanxiong Rhizome,Bidentata, Angelica Sinensis,Peach Kernel,Safflower and Red peony Root can promote blood circulation and dissipate blood stasis;Dried Rehmannia Root can nourish Yin and body fluid. Just from thecold or hot nature of the herb, Astragalus is slightly warm; White Atractylodes Rhizome, the Peel of Bitter Orange,Dried Old Orange Peel,Chuanxiong Rhizome, Angelica Sinensis and Safflower are warm; Lu Codonopsis,Balloonflower, Licorice, Peach Kernel and Bidentata are bland; Red peony Root and Bupleurum are slightly cold; Dried Rehmannia Root is cold. The quatity proportion of the slightly warm, the warm,the bland, the slightly cold and the cold is 10/42/40/11/9.The prescription has prominently warm nature and the effect of reinforcing Qi and warming Yang and activating blood circulation, and warming Yang is most important.The experimental results show that the increase in Na+-K+-ATP enzyme activity may be one of the mechanisms that cor pulmonale was treated by Decoction of Replenishing Lung and Activating Blood Circulation which was created on the thinking of reinforcing Qi and warming Yang and activating blood circulation.Conclusion:The experiment primarily revealed the treatment mechanisms of Decoction of Replenishing Lungs and Activating Blood Circulation,namely the way of YiQi,WenYang and Activating Blood Circulation to cor pulmonale, from the angle of tissue repair (promotion of cell proliferation in lung tissues),energy metabolism (enhanceing lung tissue na+k+ATP enzyme activity), vasoactive substances(increase in vascular relaxing factor NO content),anti-inflammatory effect and the side-effect(No significant differences are in TNF-αlevel between the before and after treatment and among the four groups after treatment).The experiment also denies initially the most undesirable side-effect.Selection of experimental indicators is also fully reflected the Decoction’s efficacy of strengthening healthy minerals and energy (promoting lung tissue cell proliferation, and enhancing lung tissue na+k+ATP enzyme activity),correctting the pathological situation(regulating vasoactive factors on dilation and contraction, the dissipation of thrombosis, reducing blood viscosity, etc.),and the test of Driving out pathogenic factors and sade effects (TNF%αdetection).T(e effiaacy of The Decgct)g, aA bEef profed objective, beine worthy of further study.更多还原