本文選題：黃芪水提取物 + 心肌纖維化�。� 參考：《遼寧中醫(yī)藥大學(xué)》2016年博士論文

【摘要】：目的:心肌纖維化是一種在臨床常見的慢性心血管疾病,其與動脈粥樣硬化、心肌梗死、心律失常、高血壓等眾多疾病和病理過程有著密切的關(guān)系。高血壓所致的心肌纖維化,是一種比較常見的病理表現(xiàn),現(xiàn)階段研究已發(fā)展到分子水平。高血壓誘發(fā)心肌纖維化的原因可能與炎癥、缺氧、血流動力學(xué)變化等息息相關(guān)。心肌纖維化在中醫(yī)理論中有著對應(yīng)的描述。心肌纖維化貫穿于多種疾病的病理過程中,比如高血壓、冠心病、糖尿病、心肌炎等多種病癥。上述這些病癥,在中醫(yī)理論中可以歸為“胸痹”、“心悸”、“眩暈”、“痰飲”、“水腫”等范疇。心肌纖維化的中醫(yī)病機(jī),可以概括為本虛標(biāo)實(shí)。因多屬久病勞損,病入絡(luò)脈所致,陰陽虧虛所致的本虛,表現(xiàn)為痰濁瘀阻、水濕橫生、痞塊熱毒等標(biāo)實(shí)現(xiàn)象。黃芪則是我國傳統(tǒng)中醫(yī)學(xué)中常用的一種藥材,經(jīng)過藥理學(xué)研究,已經(jīng)證實(shí)含有多種有效成分,如黃芪皂苷、黃芪多糖、黃酮類和氨基酸類等。黃芪具有增強(qiáng)機(jī)體免疫功能、保護(hù)肝臟功能、利尿、抗衰老氧化、抗應(yīng)激作用,廣譜抗菌作用,并且能夠降低血壓。傳統(tǒng)醫(yī)學(xué)的驗(yàn)方中,使用黃芪配伍其他藥物,治療相關(guān)心血管疾病的比比皆是,但其中起效的具體成分仍不清楚。黃芪水提取物是通過水提取法從黃芪的根部中提取的主要活性成分之一。本實(shí)驗(yàn)室對于黃芪水提取物的以往研究表明,黃芪水提取物具有抗炎活性,可降低巨噬細(xì)胞遷移和血管內(nèi)皮細(xì)胞的粘附,通過核轉(zhuǎn)錄因子(NF)-κB信號通路減輕局部炎癥,抗動脈粥樣硬化。但是黃芪水提取物對于高血壓相關(guān)作用,對心肌纖維化的相關(guān)影響及其機(jī)制并未見諸于報(bào)道。因此,本研究旨在通過文獻(xiàn)研究對高血壓及心肌纖維化的證型、使用藥物等作出分析,同時(shí)對本實(shí)驗(yàn)室近期所研究的藥物黃芪水提取物是否對于高血壓及其所致的心肌纖維化具有拮抗作用加以研究。材料與方法:1.文獻(xiàn)研究收集高血壓相關(guān)的研究共計(jì)3663篇,經(jīng)過篩選后入選文獻(xiàn)共527篇,通過對上述文獻(xiàn)進(jìn)行歸納,整理出高血壓相關(guān)的證型與用藥的初步總結(jié)。然后再次收集心肌纖維化相關(guān)的文獻(xiàn)490篇,通過文獻(xiàn)研究心肌纖維化的臨床治療用藥習(xí)慣來探討高血壓治療與心肌纖維化治療的共通之處,從而在臨床治療上能夠有效的對這種疾病進(jìn)行治療。2.相關(guān)實(shí)驗(yàn)研究實(shí)驗(yàn)一中的文獻(xiàn)研究顯示,高血壓比較常見的證型表現(xiàn)為肝陽上亢、痰瘀互結(jié)、肝腎陰虛等。心肌纖維化比較常見的證型包括氣虛血瘀、陽虛水泛、陰虛熱盛等證型。中藥黃芪的具有的作用非常多,包括益氣固表,利尿托毒,排膿斂瘡等,對于氣虛、陰陽兩虛等癥狀表現(xiàn)的治療作用全面,在臨床治療心肌纖維化的應(yīng)用中應(yīng)該是較為常見的一種藥材。而現(xiàn)階段,隨著藥理學(xué)與臨床醫(yī)學(xué)的發(fā)展,中藥提取物的使用逐漸成為醫(yī)學(xué)研究的后續(xù)熱潮,黃芪的提取物研究也在逐步展開,曾經(jīng)有相關(guān)報(bào)道提示,黃芪甲苷通過對內(nèi)皮的保護(hù)等,對于拮抗高血壓具有一定的作用。同時(shí),黃芪甲苷對于心肌纖維化也有一定的抑制作用。而黃芪水提取物是另一種較為常見的黃芪提取物,本實(shí)驗(yàn)室既往研究便是圍繞黃芪水提取物對內(nèi)皮因子作用進(jìn)行研究。本次的后續(xù)實(shí)驗(yàn)試圖研究,黃芪水提取物這種中藥提取物是否也一樣對于高血壓性心肌纖維化具有相關(guān)作用。2.1.試劑黃芪水提取物由中國科學(xué)院大連化學(xué)物理研究所提供,在蒸餾水中溶解并稀釋(3克/公斤/天)。辛伐他汀(simvastatin)購自sigma-aldrich(saintlouis,mo,usa)。血管緊張素Ⅱ購自美侖生物制藥(大連)。皮下埋入植入式膠囊滲透壓泵(usa/alzet生產(chǎn))。2.2.小鼠雄性昆明小鼠(n=24),年齡8周,18~20克,來源于swiss小鼠的背景,購自沈陽軍區(qū)總醫(yī)院醫(yī)學(xué)動物科。所有的小鼠全部分籠分組喂養(yǎng),以常規(guī)平價(jià)鼠糧進(jìn)行適應(yīng)性,常規(guī)性的喂養(yǎng)一周后,將小鼠按照盲法隨機(jī)分為空白對照組、血管緊張素Ⅱ模型組、血管緊張素Ⅱ加辛伐他汀組、血管緊張素Ⅱ加黃芪水提取物組(每組n=6)。除空白對照組外,其他三組全部麻醉后皮下埋入植入式膠囊滲透壓泵,內(nèi)容物為血管緊張素Ⅱ(1.3毫克/公斤/天),血管緊張素Ⅱ加辛伐他汀組小鼠每日服用0.4%辛伐他汀溶液灌胃(30毫克/公斤/天),血管緊張素Ⅱ加黃芪水提取物組小鼠給予口服劑量黃芪水提取物溶液(3克/公斤/天)。在皮下埋泵28日時(shí)收集組織標(biāo)本,進(jìn)一步準(zhǔn)備進(jìn)行數(shù)據(jù)分析。按照相關(guān)要求,所有涉及動物實(shí)驗(yàn)程序的相關(guān)實(shí)驗(yàn),都已經(jīng)獲得倫理委員會的批準(zhǔn)。2.3he染色實(shí)驗(yàn)收集到達(dá)標(biāo)點(diǎn)時(shí)間的小鼠心臟標(biāo)本,清洗干凈后投入預(yù)先配制準(zhǔn)備的固定液(一般為4%多聚甲醛)中,固定脫水后浸蠟、包塊、切片(心臟橫切,切片厚度大約5-8微米)后,將切片放入45℃恒溫箱進(jìn)行烘干步驟,通過浸泡在酒精和二甲苯中進(jìn)行脫蠟,按照步驟要求將切片投入regaud(或weigert)蘇木素溶液中染色5-10分鐘,然后投入1%鹽酸乙醇與氨水中分化數(shù)秒,使用流水沖洗大約20分鐘以上等待細(xì)胞核反藍(lán),下一步驟,使用蒸餾水反復(fù)沖洗后,將切片投入0.5%的伊紅溶液中染色約1-3分鐘,分別投入80%-100%酒精中按順序脫水后,投入二甲苯中依次序換缸,采用中性樹脂進(jìn)行封片固定,切片標(biāo)本制備后即可采用顯微鏡下觀察。2.4masson染色實(shí)驗(yàn)將小鼠的心臟標(biāo)本切片準(zhǔn)備后,投入已經(jīng)配置準(zhǔn)備的固定液(4%多聚甲醛)中,固定脫水后浸蠟、包塊、切片(心臟橫切,一般厚度5-8微米)后,放入45℃恒溫箱內(nèi)烘干,石蠟切片使用酒精進(jìn)行脫蠟、脫水,然后使用自來水和蒸餾水分別沖洗。用regaud蘇木素溶液或weigert蘇木素溶液5-10分鐘,充分進(jìn)行水洗后,觀察效果如果認(rèn)為切片過染,可以使用鹽酸酒精進(jìn)行分化,分化后再次使用蒸餾水沖洗。使用蘇木素染核結(jié)束后,使用masson麗春紅酸性復(fù)紅液浸染,大概時(shí)間為5-10分鐘,然后切片以2%冰醋酸水溶液侵染片刻,再使用1%的磷鉬酸水溶液分化進(jìn)行約3-5分鐘。不必經(jīng)過流水重新,直接用苯胺藍(lán)溶液進(jìn)行染色5分鐘,染色結(jié)束后再用0.2%冰醋酸水溶液浸洗片刻,使用80%、90%、95%酒精、無水酒精進(jìn)行脫水后,再用二甲苯進(jìn)行透明、中性樹脂封固,就可以對切片進(jìn)行鏡下觀察。2.5免疫組化技術(shù)檢驗(yàn)將手術(shù)后采集的小鼠心臟標(biāo)本清洗后,投入預(yù)先配好的固定液(4%多聚甲醛)中,固定脫水后浸蠟、包塊、切片(心臟橫切,一般厚度5-8微米)后,放入45℃恒溫箱內(nèi)烘干,石蠟切片在酒精、二甲苯中進(jìn)行脫蠟脫水,然后將切片在清水中反復(fù)沖洗一段時(shí)間,置入3%過氧化氫溶液中浸泡10分鐘,(這一步驟可以除去內(nèi)源性的過氧化氫酶)然后倒掉過氧化氫溶液,將切片放入清水中反復(fù)清洗后,取出滴加抗原修復(fù)液(本實(shí)驗(yàn)室使用為檸檬酸緩沖液),密閉放入沸水中煮40分鐘后撈出冷卻至室溫。pbs溶液沖洗后,動物血清封閉,去除動物血清添加一抗,一抗滴加完畢后,將切片盒于4℃冰箱內(nèi)過夜,第二日取出在室溫下復(fù)溫30分鐘,使用pbs溶液沖洗后在切片上添加二抗,滴加適量的辣根酶或堿性磷酸酶標(biāo)記的鏈霉卵白素工作液后,將切片用pbs溶液反復(fù)沖洗,沖洗后進(jìn)行dab染色觀察,待dab顯色達(dá)到理想程度后,清水沖洗切片以終止顯色反應(yīng),然后使用蘇木素復(fù)染染色細(xì)胞核,梯度酒精進(jìn)行脫水,二甲苯溶液透明后采用中性樹膠對切片封片固定,標(biāo)本即可觀察使用。2.6免疫印跡分析(westernblot)提取各組小鼠心臟組織,測定不同組別間的心臟組織膠原含量。本實(shí)驗(yàn)中采用westernblot檢測小鼠心臟組織的collagen-3。使用材料為100微升裂解液(50毫摩爾/升的tris,10毫摩爾/升的氯化鎂,0.5摩爾/升氯化鈉,1%tritonx-100)用來提取心肌總蛋白。心臟組織保持在冰上裂解及收取。在離心機(jī)使用14,000轉(zhuǎn)速度離心10分鐘后,將分離出來的上清收集。按照相關(guān)試劑盒上的實(shí)驗(yàn)方法提取蛋白,離心后上清保存。用bca法測定上清液中的蛋白質(zhì)水平。等分30微克蛋白質(zhì)樣品進(jìn)行聚丙烯酰胺凝膠電泳,將其轉(zhuǎn)移到聚偏二氟乙烯膜。轉(zhuǎn)膜結(jié)束后,采用5%脫脂奶粉將聚偏二氟乙烯膜封閉在tris中緩沖生理鹽水,然后在膜上添加需要的一抗,置于在4°c冰箱中,按照孵育條件過夜,次日取出后采用pbs洗滌3次,沖洗后使用1:2,000稀釋的辣根過氧化物酶(hrp)標(biāo)記的羊抗兔,羊抗鼠,兔抗山羊二抗在室溫下培養(yǎng)2小時(shí)。洗滌三次后,添加發(fā)光劑檢測活性,暗室曝光。2.7統(tǒng)計(jì)分析實(shí)驗(yàn)數(shù)據(jù)采用平均值±sd表示。所有實(shí)驗(yàn)中的數(shù)據(jù)均采用spss13.0統(tǒng)軟件統(tǒng)計(jì)結(jié)果。兩組之間的差異統(tǒng)計(jì)采用unpairedstudent’st檢驗(yàn)。三個(gè)相關(guān)實(shí)驗(yàn)組之間的差異采用方差分析(anova)。p0.05(雙尾)具有統(tǒng)計(jì)學(xué)意義。結(jié)論:1.高血壓病的證型分布以肝陽上亢型為最多、肝腎陰虛、瘀血阻絡(luò)、肝火亢盛、痰濕中阻、痰瘀互結(jié)、氣虛血瘀、陰陽兩虛、腎氣虧虛等都是較為常見的證型,此外還有多種證型存在,臨床治療要根據(jù)具體證候進(jìn)行論治。2.高血壓病的證素中以陽亢、陰虛、氣滯為最主要,臨床中出現(xiàn)頻率最高,其次為血瘀、氣虛、痰濁、火熱等。3.高血壓病的常用藥物以補(bǔ)虛藥、鎮(zhèn)肝熄風(fēng)藥、活血化瘀藥、清熱藥、理氣化痰藥為主,其他各種不同藥物根據(jù)證型加以配伍改變。4.心肌纖維化的臨床治療以活血化瘀、平肝潛陽、降濁消積、溫陽利水、寬胸散結(jié)法最為常見。現(xiàn)階段臨床治療中以活血化瘀藥、平肝熄風(fēng)藥、溫里藥、補(bǔ)虛藥、利水滲濕藥為主,通常選用復(fù)方湯劑或者其他復(fù)方制劑,中藥提取物也逐漸開始廣泛應(yīng)用于臨床。5.補(bǔ)虛類藥物在高血壓病與心肌纖維化重都有著大量的應(yīng)用。6.黃芪水提取物能夠拮抗血管緊張素Ⅱ刺激產(chǎn)生的血壓升高與心室重構(gòu),從而抑制心肌纖維化的形成。
[Abstract]:Objective: myocardial fibrosis is a common chronic cardiovascular disease, which has a close relationship with many diseases and pathological processes, such as atherosclerosis, myocardial infarction, arrhythmia, hypertension, and so on. Myocardial fibrosis caused by hypertension is a common pathological manifestation. At present, the study has developed to molecular level. The causes of myocardial fibrosis induced by blood pressure may be closely related to inflammation, hypoxia and hemodynamic changes. Myocardial fibrosis has a corresponding description in traditional Chinese medicine theory. Myocardial fibrosis runs through the pathological process of various diseases, such as hypertension, coronary heart disease, diabetes, myocarditis and many other diseases. These diseases are in Chinese medicine. The theory can be classified as "chest Bi", "palpitation", "dizziness", "phlegm" and "edema". The TCM pathogenesis of myocardial fibrosis can be summed up as a virtual standard. Because of many chronic diseases, the disease entered the collateral veins, the deficiency of yin and Yang caused by the deficiency of phlegm and blood stasis, the water wet cross, the heat poison of the lump, and so on. A kind of medicinal herbs commonly used in traditional Chinese medicine has proved to contain a variety of effective components, such as Astragalus saponins, astragalus polysaccharides, flavonoids and amino acids. Astragalus membranaceus can enhance the immune function of the body, protect liver function, diuresis, anti aging oxidation, anti stress effect, broad-spectrum antibacterial effect, and can reduce blood pressure. In traditional medicine, the use of Astragalus and other drugs to treat related cardiovascular diseases is abound, but the specific components of the effect are still unclear. The water extract of Astragalus is one of the main active ingredients extracted from the root of Astragalus membranaceus through water extraction. The extract of Astragalus membranaceus has anti-inflammatory activity, which can reduce the migration of macrophages and the adhesion of vascular endothelial cells, alleviate local inflammation and anti atherosclerosis through the nuclear factor (NF) - kappa B signaling pathway, but the effect of Astragalus membranaceus water extract on hypertension and the related effects on myocardial fibrosis and its mechanism are not reported. The purpose of this study is to analyze the evidence of hypertension and myocardial fibrosis by literature, and to analyze the antagonistic effects of Astragalus membranaceus water extract on hypertension and myocardial fibrosis in recent years. Materials and methods: 1. literature studies are related to the collection of hypertension. A total of 3663 articles were selected and 527 articles were selected after screening. Through the induction of the above literature, a preliminary summary of hypertension related syndromes and drugs was sorted out. Then 490 articles related to myocardial fibrosis were collected again. The treatment and heart of hypertension in the treatment and heart of hypertension were studied through literature Study on the habit of treating myocardial fibrosis in the treatment of bed. The common place of the treatment of muscle fibrosis, so as to be effective in the treatment of the disease in clinical treatment, the literature study in the experimental study of.2. related experiments shows that the common syndromes of hypertension are liver yang hyperactivity, phlegm and blood stasis, deficiency of liver and kidney, etc. the common syndromes of myocardial fibrosis include Qi deficiency, blood stasis and yang deficiency. The effect of traditional Chinese medicine Astragalus membranaceus is very much, including Qi Qi and fixation, diuresis, purulent and collecting sores, etc., in the treatment of qi deficiency, yin and yang two deficiency symptoms and other symptoms, in the clinical treatment of myocardial fibrosis should be a more common medicine. At the present stage, with pharmacology and clinical medicine The use of Chinese medicine extracts has gradually become a subsequent upsurge of medical research. The study on the extract of Astragalus has also been carried out gradually. It has been reported that astragaloside has a certain effect on antagonizing hypertension through the protection of endothelium and so on. At the same time, astragaloside also has certain inhibitory effect on myocardial fibrosis. The Astragalus membranaceus extract is a more common extract of Astragalus membranaceus. The previous study in our laboratory was to study the effect of Huang Qishui extract on the endothelial factor. This follow-up experiment tried to study whether the extract of Astragalus membranaceus water extract was also the same as the.2.1. test for high blood pressure myocardial fibrosis. The aqueous extract of Astragalus membranaceus was provided by the Dalian Institute of Chemical Physics, Chinese Academy of Sciences, dissolved and diluted in distilled water (3 grams per kilogram / day). Simvastatin was purchased from Sigma-Aldrich (saintlouis, Mo, USA). Angiotensin II was purchased from MLUN biopharmaceutical (Dalian). Subcutaneous embedded capsule osmotic pressure pump (usa/alzet production).2.2. The male mouse Kunming mice (n=24), aged 8 weeks, 18~20 gram, derived from the background of Swiss mice, were purchased from the Medical Animal Department of the General Hospital of Shenyang military region. All the mice were fed in all the cage groups and were fed with conventional parity rats. After a week of routine feeding, the rats were randomly divided into blank control groups according to the blind method, angiotensin II model. Group, angiotensin II plus simvastatin group, angiotensin II Plus Astragalus water extract group (each group of n=6). Except for the blank control group, all the other three groups were subcutaneously embedded into the implanted capsule osmotic pressure pump, the contents were angiotensin II (1.3 mg / kg / day), and angiotensin II plus simvastatin group took 0.4% daily. Simvastatin solution (30 mg / kg / day), angiotensin II and Astragalus membranaceus water extract group were given oral dose of Astragalus membranaceus water extract solution (3 g / kg / day). Tissue specimens were collected at 28 days of subcutaneous embedded pump to further prepare for data analysis. All related experiments involving animal experiment procedures were conducted according to the relevant requirements. It has been approved by the ethics committee's approval.2.3he staining experiment to collect the heart specimens of the mice that arrive at the punctuation time and put into the pre prepared fixed liquid (usually 4% polyformaldehyde) after cleaning. After dehydration, paraffin, lumps, slices (heart crosscutting, thickness about 5-8 microns), the slices are put into a thermostat at 45 degrees centigrade to be baked. Dry steps, dewaxing by soaking in alcohol and dimethylbenzene, dyeing the slice into regaud (or Weigert) soubin solution for 5-10 minutes according to the steps, and then putting into 1% hydrochloric acid ethanol and ammonia water for several seconds, and using water for more than 20 minutes to wait for the nucleus anti blue for more than 20 minutes, and the next step, using distilled water to rinse again and again. After putting the slice into 0.5% eosin solution for about 1-3 minutes, then putting into 80%-100% alcohol in order to dehydrate in sequence, then put into the dimethylbenzene in order to change the cylinder in order, and use the neutral resin to fix the seal. After the preparation of the slice specimen, the specimen of the heart of the mice can be observed under the microscope and then the specimens of the mice are prepared and put into the section. After the fixed solution (4% polyformaldehyde), after the fixed dehydration, the wax is dipped, the block, the slice (the heart crosses, the general thickness is 5-8 microns), then dried in the constant temperature box at 45 C, and the paraffin section is dewaxed and dehydrated using alcohol. Then, the water and the distilled water are used respectively. 5-10 points are used with regaud hematoxylin solution or Weigert hematoxylin solution. When the clock is washed fully, the observation effect, if it is considered that the slice is dyed, can be differentiated by hydrochloric acid and then flushed with distilled water after differentiation. After the use of hematoxylin to dye the nucleus, the Masson Li Chun Hong acid reddish liquid is used for 5-10 minutes, and then the slice is infected by 2% glacial acetic acid solution for a moment, and then use 1%. The solution of Phospho molybdate water is divided for about 3-5 minutes. Without water, it will be dyed for 5 minutes directly with aniline blue solution. After the dyeing, it is soaking for a moment with 0.2% glacial acetic acid solution, using 80%, 90%, 95% alcohol, and anhydrous alcohol to dehydrate, then dimethylbenzene is transparent, and the neutral resin is sealed. After cleaning the specimens of the mice after the operation, after the.2.5 immunohistochemical technique, they were put into the pre matched fixed solution (4% polyformaldehyde). After the dehydration was fixed, the wax was impregnated, the paraffin block and the slice (the heart crosscutting, the general thickness of 5-8 microns) were dried in a constant temperature box at 45 C, and paraffin section was dewaxed and dehydrated in alcohol and xylene. Then, the paraffin section was dewaxed and dehydrated. Then, the paraffin section was dewaxed and dehydrated. The slice was rinsed in clear water for a period of time and was soaked in 3% hydrogen peroxide solution for 10 minutes. (this step could remove the endogenous catalase) and then pour out the hydrogen peroxide solution. After rinsing the slice into the clean water, the antigen repair solution was extracted (the laboratory was used as a citric acid buffer) and closed into the boiling water. After 40 minutes of water boiled in the water and cooled to room temperature.Pbs solution, the animal serum was closed and the animal serum was added to add a resistance. After the first anti drop, the sliced box was taken overnight in the refrigerator at 4, and the temperature was reheated at room temperature for second days for 30 minutes. After washing with the PBS solution, the slices were added to the slice and added a proper amount of horseradish or alkaline phosphoric acid. After the enzyme labelled streptomycin working fluid, the slice was rinsed with PBS solution repeatedly and followed by DAB staining. After the DAB coloration reached the ideal degree, the clear water was flushed to terminate the color reaction. Then the cell nucleus was stained with hematoxylin and the gradient alcohol was used to dehydrated, and the dimethylbenzene solution was transparent with neutral gum after the slice. .2.6 immunoblotting analysis (Westernblot) was used to observe the cardiac tissue of each group and determine the collagen content of cardiac tissue in different groups. In this experiment, the use of Westernblot to detect collagen-3. in the cardiac tissue of mice was 100 microlitre solution (50 mole / liter Tris, 10 mole / liter chlorine). Magnesium, 0.5 mole / liter of sodium chloride, 1%tritonx-100) used to extract total myocardial protein. Cardiac tissue remains frozen and collected on ice. The isolated supernatant is collected after 14000 rotation centrifugation in centrifuge for 10 minutes. The protein is extracted according to the experimental method on the related kit and preserved in the supernatant after centrifugation. The supernatant is determined in the supernatant by BCA method. Protein level. 30 microgram protein samples were divided into polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride (pvvdf) membrane. After the film was finished, 5% skimmed milk powder was used to close the polyvinylidene fluoride membrane in Tris Physiological Buffered Saline, and then the required first resistance was added to the membrane. It was placed in the 4 degree C refrigerator and spent the night according to the incubation conditions. After taking out the next day, 3 times were washed with PBS. After rinsing, the Sheep anti rabbit, Sheep anti rat and Rabbit anti goat two anti goat were cultured with 1:2000 diluted horseradish peroxidase (HRP) for 2 hours at room temperature. After three times washing, the activity was detected by adding luminescent agent. The statistical analysis data of dark room exposure were expressed by mean value of + SD. All the data in the experiment The statistical results of SPSS13.0 software were used. The difference between the two groups was measured by unpairedstudent 'st test. The difference between the three related experimental groups was statistically significant by using variance analysis (ANOVA).P0.05 (double tail). Conclusion: the syndrome type distribution of 1. hypertension is the most in liver yang hyperactivity, liver kidney yin deficiency, blood stasis obstructing collaterals, liver fire hyperactivity, The obstruction of phlegm dampness, phlegm and blood stasis, Qi deficiency and blood stasis, yin and yang two deficiency and kidney qi deficiency are common syndrome types. In addition, there are many kinds of syndrome types. The clinical treatment should be based on specific syndromes to treat.2. hypertension with yang hyperactivity, yin deficiency and qi stagnation as the most important, the second is blood stasis, Qi deficiency, phlegm turbidity, and heat. The commonly used drugs for.3. hypertension are to fill in the deficiency drugs, the antipyretic drug of the liver, the medicine of activating blood and removing stasis, the antipyretic medicine and the medicine of Qi and chemical phlegm, and the other different drugs are combined with the syndrome type to change the clinical treatment of.4. myocardial fibrosis with the treatment of activating blood and removing stasis, smoothing the liver and Yang, reducing turbidity and eliminating the accumulation, warming Yangli water, and the broad chest disintegration. In the treatment, the drugs of promoting blood circulation and removing blood stasis, the medicine of extinguishing the liver, the medicine of warm water, the medicine of filling the deficiency, the medicine of the water and the dampness, usually use the compound decoction or other compound preparation. The extract of the Chinese medicine has gradually started to be widely used in the clinical.5. supplement deficiency drugs in the hypertension and the myocardial fibrosis, and the.6. Astragalus water extract can antagonize the blood. Angiotensin II stimulates increased blood pressure and ventricular remodeling, thereby inhibiting the formation of myocardial fibrosis.

【學(xué)位授予單位】：遼寧中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R285.5

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