發(fā)布時(shí)間：2018-05-04 09:48

  本文選題：動(dòng)脈粥樣硬化 + 膽固醇逆向轉(zhuǎn)運(yùn)��； 參考：《北京中醫(yī)藥大學(xué)》2015年博士論文

【摘要】：動(dòng)脈粥樣硬化(AS)是導(dǎo)致心腦血管疾病的重要因素,目前AS引起的心血管疾病(CVD)仍居發(fā)達(dá)國(guó)家人口死亡原因之首。人民壽命的增長(zhǎng),人口老齡化的出現(xiàn),使得AS發(fā)病率、致死率居高不下。因此,對(duì)于AS的發(fā)病原因、發(fā)病機(jī)制、預(yù)防及治療措施一直是心血管病研究者的研究熱點(diǎn)。AS是由多種因素共同作用引起的多系統(tǒng)參與疾病,作用機(jī)制復(fù)雜,故而單一作用機(jī)制的藥物無法起到有效抗AS的作用。中藥復(fù)方以中醫(yī)理論為指導(dǎo),以整體觀為主導(dǎo)思想,著眼于宏觀、綜合治療,從多角度、多層次、多靶點(diǎn)有效地防治AS,是目前抗AS研究的熱點(diǎn)�！懊}濁”理論的提出,豐富了中醫(yī)學(xué)對(duì)AS病機(jī)的認(rèn)識(shí)角度,并將中醫(yī)學(xué)與現(xiàn)代醫(yī)學(xué)相通,化濁通脈法正是針對(duì)這一證候所創(chuàng)立,以此為治法的中藥復(fù)方在前期臨床觀察中取得了良好的療效。目的：觀察化濁通脈方對(duì)AS兔主動(dòng)脈血管壁AS斑塊形成以及對(duì)血清和主動(dòng)脈血管壁細(xì)胞內(nèi)脂質(zhì)含量的影響,從其影響膽固醇逆向轉(zhuǎn)運(yùn)(RCT)過程的作用,探討化濁通脈方抗AS的作用機(jī)制。實(shí)驗(yàn)方法：健康純種雄性新西蘭大耳白兔40只隨機(jī)分為4組：空白對(duì)照組,模型組,辛伐他汀組,化濁通脈方組(中藥復(fù)方組)。采用單純高脂飼料喂養(yǎng)方式制備AS兔模型,分別在給藥前及給藥14周末禁食后經(jīng)耳緣靜脈采集血液樣本,給藥15周末經(jīng)耳緣靜脈麻醉后處死動(dòng)物,取胸主動(dòng)脈及肝臟樣本。肉眼觀察各組兔胸主動(dòng)脈標(biāo)本脂紋、脂斑形成情況；主動(dòng)脈標(biāo)本行HE染色、油紅O染色,觀察各組兔主動(dòng)脈壁脂質(zhì)沉積及斑塊形成情況；酶法測(cè)定血清中總膽固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)水平；酶聯(lián)免疫吸附法(ELISA)測(cè)定主動(dòng)脈血管壁細(xì)胞內(nèi)TC、游離膽固醇(FC)水平,肝臟細(xì)胞內(nèi)CD36水平；蛋白印跡法(WB)測(cè)定主動(dòng)脈壁內(nèi)小凹蛋白-1(cav-1).親環(huán)素A、三磷酸腺苷結(jié)合盒轉(zhuǎn)運(yùn)蛋白Al (ABCA1)、肝臟B類I型清道夫受體(SR-B I)蛋白表達(dá)量；實(shí)時(shí)定量聚合酶鏈?zhǔn)椒磻?yīng)(real-time PCR)測(cè)定主動(dòng)脈壁cav-1 mRNA.親環(huán)素A mRNA、ABCA1 mRNA.肝臟SR-B I mRNA. CD36 mRNA表達(dá)量。實(shí)驗(yàn)結(jié)果：空白對(duì)照組兔主動(dòng)脈血管壁內(nèi)膜表面光滑,各層結(jié)構(gòu)清晰,內(nèi)皮細(xì)胞完整連續(xù),平滑肌細(xì)胞排列整齊緊密有序,無脂質(zhì)、斑塊等沉積；模型組兔主動(dòng)脈血管壁明顯增厚,內(nèi)皮層脫落,管壁滿布脂質(zhì)、斑塊；中藥復(fù)方組兔主動(dòng)脈壁內(nèi)膜稍有增厚,內(nèi)膜下可見散在泡沫細(xì)胞沉積及脂質(zhì)沉積,未見明顯斑塊,與辛伐他汀組無明顯差異。14周末與空白對(duì)照組相比,模型組兔血清中TC、TG、LDL顯著升高(P0.01),平滑肌細(xì)胞內(nèi)TC、FC明顯升高(P0.05,P0.01)；與模型組相比,中藥復(fù)方組兔血清中TC、TG、LDL明顯下降(P0.05,P0.01),主動(dòng)脈血管壁細(xì)胞內(nèi)TC、FC顯著升高(P0.01)。15周末主動(dòng)脈細(xì)胞測(cè)定結(jié)果顯示：與空白對(duì)照組相比,模型組兔cav-1蛋白及mRNA表達(dá)量顯著下降(P0.01)；與模型組相比,中藥復(fù)方組cav-1 mRNA表達(dá)量顯著增加(P0.01),cav-1蛋白表達(dá)量也有增加趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義(P0.05)。與空白對(duì)照組相比,模型組兔親環(huán)素A蛋白表達(dá)量明顯增加(P0.05),親環(huán)素A mRNA表達(dá)量有下降趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義(P0.05)；與模型組相比,中藥復(fù)方組親環(huán)素A蛋白表達(dá)量顯著下降(P0.01),親環(huán)素A mRNA表達(dá)量顯著增加(P0.01)。與空白對(duì)照組相比,模型組兔ABCA1蛋白表達(dá)量有下降趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義(P0.05),ABCA1 mRNA表達(dá)量顯著增加(P0.01)；與模型組相比,中藥復(fù)方組ABCA1蛋白表達(dá)量有增加趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義P0.05), ABCA1 mRNA表達(dá)量顯著下降(P0.01)。肝細(xì)胞測(cè)定結(jié)果顯示：與空白對(duì)照組相比,模型組兔SR-B I蛋白及mRNA表達(dá)量有下降趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義(P0.05)；與模型組相比,中藥復(fù)方組SR-B I mRNA表達(dá)量有增加趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義(P0.05)。與空白對(duì)照組相比,模型組兔CD36蛋白表達(dá)量有增加趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義P0.05), CD36 mRNA表達(dá)量明顯增加(P0.05)；與模型組相比,中藥復(fù)方組CD36蛋白表達(dá)量有減少趨勢(shì),但差異無統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論：1.化濁通脈方能夠抑制AS兔主動(dòng)脈血管內(nèi)膜增生,減少內(nèi)皮下泡沫細(xì)胞的形成及脂質(zhì)沉積,從而減少AS斑塊形成。2.化濁通脈方可有效降低AS兔血清TC、TG、LDL水平,顯著降低AS兔主動(dòng)脈血管壁細(xì)胞內(nèi)TC、FC含量。3.化濁通脈方能夠通過提高cav-1蛋白及基因水平、親環(huán)素A的基因水平,促進(jìn)細(xì)胞內(nèi)膽固醇轉(zhuǎn)運(yùn)復(fù)合體生成,對(duì)ABCA1蛋白表達(dá)量有上調(diào)作用趨勢(shì),促進(jìn)細(xì)胞內(nèi)膽固醇流出,保障cav-1、親環(huán)素A的正常表達(dá),減少斑塊中親環(huán)素A的過度表達(dá),最終達(dá)到抗AS的作用。4.化濁通脈方能夠通過對(duì)肝臟SR-B I基因水平有一定的上調(diào)作用趨勢(shì)、對(duì)CD36蛋白表達(dá)有一定的抑制作用趨勢(shì),可能促進(jìn)血中膽固醇被肝臟選擇性攝取、代謝,使脂質(zhì)順利代謝,最終達(dá)到抗AS的作用。
[Abstract]:Atherosclerosis (AS) is an important factor leading to cardiovascular and cerebrovascular diseases. At present, the cardiovascular disease (CVD) caused by AS is still the first cause of death in the developed countries. The growth of the people's life and the emergence of the aging population make the incidence of AS and death rate high. Therefore, the causes, pathogenesis, prevention and treatment measures for the pathogenesis of AS are made. The research hotspot of cardiovascular disease researchers.AS is a multi system involved in disease caused by a variety of factors. The mechanism is complex, so the single action mechanism can not play an effective anti AS effect. Chinese medicine compound is guided by the theory of traditional Chinese medicine, guided by the concept of holism, focusing on macroscopical, comprehensive treatment, from multi angle. Degree, multi-level and multi target effectively prevent and cure AS, is the hot spot of anti AS research at present. "Pulse turbid" theory has enriched the understanding angle of Chinese medicine to AS pathogenesis, and interlinked the traditional Chinese medicine with modern medicine. Objective: To observe the effect of Hua Zhuo Tong Mai Recipe on the formation of AS plaque in the aorta wall of AS rabbit and the lipid content in the blood serum and aorta wall cells, and to explore the mechanism of anti AS in the process of RCT. The method of actual test: healthy pure breed male New Zealand white rabbit 40 Only 4 groups were randomly divided into 4 groups: blank control group, model group, simvastatin group, turbid Tongmai prescription group (Chinese medicine compound group). The rabbit model was prepared by feeding simple high fat diet. The blood samples were collected before and after the 14 weekend fasting, and the animals were killed after the ear vein anesthesia at the end of the 15 week and the thoracic aorta was taken for the 15 week. The lipid profiles and formation of fat spots in the thoracic aorta of rabbits were observed by the naked eye. The lipid deposition and plaque formation of aorta wall in each group were observed by HE staining and oil red O staining. The serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL) and high density lipoprotein (HDL) were measured by enzyme method. Enzyme-linked immunosorbent assay (ELISA) was used to determine the intracellular TC, free cholesterol (FC) level, CD36 level in the liver cells, and Western blot (WB) for the determination of -1 (Cav-1) in the aortic wall. The expression of cyclophilin A, adenosine triphosphate binding cassette transporter Al (ABCA1), and liver B I type scavenger receptor (Cav-1) protein expression; The quantitative polymerase chain reaction (real-time PCR) was used to determine the Cav-1 mRNA. Pro cyclophilin A mRNA of the aortic wall and the SR-B I mRNA. CD36 expression of the ABCA1 mRNA.. The experimental results showed that the intima surface of the aorta wall of the rabbit aorta was smooth, the structure of each layer was clear, the endothelial cells were complete and continuous, the smooth muscle cells were arranged neatly and orderly, without lipid. In the model group, the aorta wall of the rabbit aorta was thickened obviously, the endothelial layer fell off, the wall of the tube was covered with lipid and plaque. The intima of the aorta was slightly thickened in the compound group of Chinese medicine, and the deposition of foam cells and lipid deposition were found under the intima, and there was no obvious plaque in the intima. The.14 weekend was compared with the blank control group, compared with the blank control group. The serum levels of TC, TG and LDL in the rabbit serum were significantly increased (P0.01), and TC and FC in the smooth muscle cells were significantly increased (P0.05, P0.01). Compared with the model group, the serum TC, TG, LDL in the rabbit serum of the Chinese medicine compound group decreased significantly (P0.05, P0.01), and the aortic vascular wall cells in the aorta were significantly higher than those in the blank control group. The expression of Cav-1 protein and mRNA in the model group decreased significantly (P0.01). Compared with the model group, the expression of Cav-1 mRNA in the compound group of Chinese medicine increased significantly (P0.01), and the expression of Cav-1 protein also increased, but the difference was not statistically significant (P0.05). Compared with the blank control group, the expression of cyclophilin A protein in the model group was significantly increased (P0.05), pro ring. There was no significant difference in the expression of A mRNA, but there was no significant difference (P0.05). Compared with the model group, the expression of Pro cyclophilin A protein decreased significantly (P0.01), and the expression of cyclophilin A mRNA increased significantly (P0.01). Compared with the blank control group, the expression of ABCA1 protein in the model group had a downward trend, but the difference was not statistically significant (P0.0). (P0.0, the difference was not statistically significant (P0.0). 5), the expression of ABCA1 mRNA increased significantly (P0.01). Compared with the model group, the expression of ABCA1 protein in the Chinese medicine compound group increased, but the difference was not statistically significant P0.05), and the expression of ABCA1 mRNA decreased significantly (P0.01). The results of hepatocyte determination showed that the expression of SR-B I protein and mRNA in the model group had a decreasing trend compared with the blank control group. But the difference was not statistically significant (P0.05). Compared with the model group, the expression of SR-B I mRNA in the Chinese medicine compound group increased, but the difference was not statistically significant (P0.05). Compared with the blank control group, the expression of CD36 protein in the model group was increased, but the difference was not statistically significant P0.05), CD36 mRNA expression increased significantly (P0.05); and the model of the model group (P0.05); The expression of CD36 protein in the Chinese medicine compound group decreased, but the difference was not statistically significant (P0.05). Conclusion: 1. Hua Tong Mai Fang can inhibit the intima hyperplasia of aorta in AS rabbit, reduce the formation of the foam cells under the endothelium and the lipid deposition, and reduce the form of AS plaques form.2. turbid and Tongmai, which can effectively reduce the TC, TG, LDL in the serum of AS rabbit. Level, significantly reduced TC in AS rabbit aortic vascular wall cells, FC content.3. turbid pulse can improve the level of Cav-1 protein and gene, the gene level of cyclophilin A, promote the formation of intracellular cholesterol transport complex, the upward trend of the expression of ABCA1 protein, promote intracellular cholesterol outflow, guarantee Cav-1, and procyclin A Normal expression, reduce the overexpression of cyclophilin A in plaque, and finally achieve the anti AS effect of.4. turbid vein, which can have a certain trend of up regulation of the liver SR-B I gene, and can inhibit the expression of CD36 protein, which may promote the selective uptake and metabolism of cholesterol in the liver and the smooth metabolism of the lipid. Finally, the effect of anti AS is achieved.

【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R285.5

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