从对HDL亚型分布及功能的影响探讨芎芍胶囊抗动脉粥样硬化的作用机制
发布时间：2018-08-02 11:33
【摘要】：动脉粥样硬化是多种心脑血管疾病的病理基础。动脉粥样硬化的发病机制主要有血脂代谢异常、炎症反应、内皮损伤、氧化应激等。高密度脂蛋白(High density lipoprotein, HDL)能够通过胆固醇逆转运(Reverse cholesterol transport, RCT)、抗炎、抗氧化、抗血栓、抗细胞凋亡及舒张血管来实现保护血管、抗动脉粥样硬化的功能。而最新的研究发现单纯HDL水平不能反映其抗动脉粥样硬化功能,而HDL的功能与其亚型、代谢、组成成分密切相关。HDL的亚型分布、组分和功能开始成为目前抗动脉粥样硬化研究的热点。ApoA-Ⅰ是HDL的主要组成蛋白和结构基础,也是HDL完成RCT、抗内皮细胞凋亡、抗氧化、抗炎功能的主要承担者。ApoA-Ⅰ数量减少或结构改变,HDL原有功能则会减弱或丧失,甚至出现促AS作用。HDL经由preβ1-HDL→HDL3→HDL2的过程而逐渐成熟,HDL2作为大的成熟HDL亚型颗粒,可促使胆固醇向肝脏和合成类固醇激素的组织转运,低HDL2水平与冠心病的风险负相关。三磷酸腺苷结合盒转运子A1(ATP-binding cassette transporters A1, ABCA1)能够与HDL结合,促进细胞内胆固醇的流出,影响HDL颗粒生成,参与RCT,调节脂质代谢。此外,ABCA1还可通过抑制炎症因子表达、参与氧化应激反应等多种途径影响AS进程。B类Ⅰ型清道夫受体(Scavenger receptor class B, type Ⅰ, SR-BI)能够选择性摄取HDL中的胆固醇酯,传递给肝脏和类固醇激素生成组织,完成RCT；还能够介导外周细胞胆固醇流出过程,参与多种脂蛋白的代谢。髓过氧化物酶(Myeloperoxidase, MPO)和对氧磷酶1(Paraoxonase 1,PON1)是HDL影响炎症、氧化应激的相关蛋白。MPO能够选择性氧化修饰ApoA-Ⅰ,使HDL功能减弱或丧失,加速AS斑块发展。相反,PON1能够直接参与脂蛋白中过氧化物的水解,使HDL免受氧化修饰,保护HDL的抗氧化功能。卵磷脂胆固醇酰基转移酶(Lecithin cholesterol acyltransf erase, LCAT)可以将胆固醇酯化,使胆固醇不断进入到HDL中,使HDL逐渐变成富含胆固醇酯的成熟HDL。LCAT是HDL代谢的关键酶,当LCAT的功能受损时,胆固醇酯的合成会受到抑制,从而导致高胆固醇血症：同时,HDL成熟过程将会受阻,AS的发生率将增加。中医认为血脂异常是内因与外因互相作用的结果,内因方面主要是脾失健运、肝失疏泄、肾精亏虚,外因包括饮食不节、情志失调、劳逸失宜等。病机方面,认为本病本虚标实,以正虚为本,湿浊、痰凝、瘀血为标,脾肝肾三脏功能失调是产生血脂异常的主要病理基础。治疗方面,辨证论治,主要以疏肝理气、滋养肝肾、健脾消食,以及活血祛瘀、化痰通络为治则。已经证实多种中药单体成分、单味中药及中药复方能够调节血脂。很多报道提示中药能够提高HDL水平,但是目前对调脂中药通过调节HDL亚型分布来影响其功能方面研究不足,而且少有从影响HDL抗炎、抗氧化功能出发研究中药抗动脉粥样硬化机制的研究报道。在前期实验中我们发现活血化瘀药芎芍胶囊抗AS作用确切,AS兔中血脂TC、LDL升高时伴有HDL升高,考虑HDL具有异质性,HDL水平升高并不一定能够抗AS。在前期研究的基础上,本次实验围绕HDL的亚型、代谢、组分和功能展开了进一步研究。目的：本研究通过建立兔动脉粥样硬化模型,从调节脂质代谢、影响HDL亚型分布及功能的角度探讨芎芍胶囊抗AS可能的作用机制。方法：1.分组和给药将60只雄性新西兰兔随机分为5组,空白对照组、模型组、辛伐他汀组、芎芍低剂量组、芎芍高剂量组,各12只。采用单纯高脂饲料喂养法建立兔AS模型。给药方法：①空白对照组予普通饲料喂养22周；②模型组予高脂饲料喂养14周,接着普通饲料喂养8周；③辛伐他汀组予高脂饲料及辛伐他汀喂养14周,后8周喂养普通饲料及辛伐他汀,辛伐他汀给药量为2mg/Kg·d；④芎芍低剂量组予高脂饲料及中药喂养14周,后8周喂普通饲料及中药,中药剂量为川芎1.5g/kg·d、赤芍0.75g/kg·d;⑤芎芍高剂量组予高脂饲料及中药喂养14周,后8周喂普通饲料及中药,中药剂量为川芎3.0g/kg·d、赤芍1.5g/kg·d。2.病理组织学观察主动脉斑块形成情况22周末麻醉之后处死动物,取出胸主动脉,肉眼观察主动脉大体标本血管壁脂质斑块形成情况。用中性福尔马林溶液固定,常规制成组织石蜡切片,苏木素-伊红(HE)染色,显微镜下观察其组织病理改变。3.高密度脂蛋白及其他血脂检测分别于实验前、给药14周、给药22周3个时间点采血,离心后用全自动生化分析仪检测血清HDL及其组分ApoA-Ⅰ、总胆固醇(TC)、低密度脂蛋白(LDL)、载脂蛋白B(ApoB)、甘油三酯(TG)、极低密度脂蛋白(VLDL)水平。4.高密度脂蛋白亚型检测取血清标本,采用酶联免疫吸附试验法(ELISA)测定血清中HDL亚型HDL2的水平。5.胆固醇逆转运功能检测实验末取肝组织于液氮中冻存。用Trizol法提取肝脏总RNA,实时荧光定量聚合酶链式反应(real-time PCR)测定肝脏ABCAl mRNA、SR-BI mRNA的表达量。6.高密度脂蛋白抗氧化功能的检测采用邻连茴香胺法测定血清MPO活性,酶联免疫吸附试验法(ELISA)测定血清PON1活性。7.高密度脂蛋白代谢调节采用实时荧光定量聚合酶链式反应(real-time PCR)测定肝脏LCAT mRNA的表达量。结果：1.空白对照组兔主动脉管壁表面光滑,内皮细胞连续,未见脂质沉积；模型组兔主动脉管壁满布脂质斑块,内膜下可见大量堆积的泡沫细胞,平滑肌层细胞内见大量脂质沉积；各给药组兔主动脉管壁表面脂质斑块较模型组减少,显微镜下见内膜下泡沫细胞堆积较少。2.给药及造模14周,除空白对照组外,各组实验兔的血清TC、TG、HDL, LDL、VLDL、ApoA-I、ApoB均升高,差异有统计学意义(P0.05或P0.01)；给药22周,除空白对照组外,各组实验兔的血清TC、VLDL、ApoA-I较给药前升高,模型组LDL较给药前升高,差异有统计学意义(P0.05或P0.01)。给药及造模14周,模型组血清TC、TG、HDL、LDL、VLDL、ApoA-I、ApoB较空白对照组升高,差异有统计学意义(P0.05或P0.01),给药组与模型组差异无统计学意义。给药22周,模型组血清TC、LDL、VLDL、ApoA-I较空白对照组升高,差异有统计学意义(P0.01);给药组血清HDL的变化与模型组相比无明显差异；给药组血清TC、LDL、VLDL较模型组降低,ApoA-I较模型组升高,差异有统计学意义(P0.05或P0.01)；其中血清TC、VLDL、ApoA-I在芎芍低、高剂量组与辛伐他汀组之间无明显差异。3.给药22周,模型组血清HDL2较空白对照组升高,差异有统计学意义(P0.05)；辛伐他汀组和芎芍高剂量组血清HDL2的升高程度较模型组更大,差异有统计学意义(P0.05)。4.给药22周末,模型组ABCA1 mRNA表达量较空白对照组升高(P0.05)。给药组ABCA1 mRNA表达量较模型组明显升高(P0.01)。模型组SR-BI mRNA表达量较空白对照组明显升高(P0.01)。各给药组SR-BI mRNA表达量与模型组相比无明显统计学差异。5.造模及给药14周后,模型组血清MPO活性较空白对照组明显升高(P0.01),各治疗组MPO活性较模型组降低(P0.05)。给药22周后,模型组MPO活性较空白对照组升高,差异有统计学意义(P0.05)；各给药组MPO较模型组降低,差异有统计学意义(P0.05)。各组实验兔血清PON1活性改变无差异(P0.05)。6.给药22周末,模型组LCAT mRNA表达量较空白对照组明显升高(P0.01)。各给药组LCAT mRNA表达量较模型组明显升高(P0.01)。结论：1.芎芍胶囊能够抑制AS兔主动脉斑块形成,减轻脂质在血管内壁的沉积,减少泡沫细胞在内膜下的聚集。2.芎芍胶囊能够升高AS兔血清ApoA-I水平,降低TC、LDL、VLDL水平,调节脂质水平。3.芎芍胶囊能够升高AS兔血清中HDL2水平,增加HDL成熟亚型颗粒的水平,影响HDL亚型分布。4.芎芍胶囊能够上调AS兔肝脏ABCA1 mRNA表达,促进肝脏内胆固醇代谢,促进胆固醇逆转运,其对肝脏SR-BI mRNA的调节作用不明显。5.芎芍胶囊能够降低AS兔血清MPO活性,抑制其氧化HDL中的ApoA-I,保护HDL抗氧化功能,其对血清PON1活性作用不明显。6.芎芍胶囊能够上调AS兔肝脏LCAT mRNA表达,促进HDL的成熟,影响HDL的功能。7.芎芍胶囊抗动脉粥样硬化的机制可能与促进HDL成熟、增加HDL成熟亚型颗粒水平以及调节血脂水平有关。8.芎芍胶囊抗动脉粥样硬化的机制可能与增加RCT相关蛋白基因表达促进RCT,且可能保护HDL抗氧化功能有关。
[Abstract]:Atherosclerosis is the pathological basis of various cardiovascular and cerebrovascular diseases. The pathogenesis of atherosclerosis is mainly lipid metabolism, inflammatory reaction, endothelial injury, oxidative stress, etc. High density lipoprotein (HDL) can pass cholesterol reverse transport (Reverse cholesterol transport, RCT), anti-inflammatory, antioxidant, Antithrombotic, anti apoptotic and diastolic blood vessels to protect blood vessels and anti atherosclerotic functions. The latest research has found that the level of HDL alone does not reflect its anti atherosclerotic function, and the function of HDL is closely related to the subtypes of subtypes, metabolism, and components of the.HDL, and the components and functions of the subtypes and functions are now becoming the present anti porridge. .ApoA- I, a hot spot in the study of sample sclerosis, is the main component of the protein and structure of HDL, and is also the main bearer of HDL to complete RCT, anti endothelial cell apoptosis, anti-oxidation, and anti-inflammatory function, the number of.ApoA- I decrease or structural change, the original function of HDL will be weakened or lost, and even the AS action.HDL via pre beta 1-HDL to HDL3 to HDL2. Gradually mature, HDL2, as a large mature HDL subtype, can promote the transport of cholesterol to the liver and the tissue of the synthetic steroid hormone, and the low HDL2 level is negatively related to the risk of coronary heart disease. The adenosine triphosphate binding cassette transporter A1 (ATP-binding cassette transporters A1, ABCA1) can be combined with HDL to promote intracellular cholesterol efflux. The effect of HDL particles formation, participation in RCT, regulating lipid metabolism. In addition, ABCA1 can also inhibit the expression of inflammatory factors and participate in the oxidative stress response in many ways, such as the AS process.B class type I scavenger receptor (Scavenger receptor class B, type I, SR-BI) can selectively absorb cholesterol esters in HDL, transmitted to liver and steroid irrigations The hormone producing tissue, completing RCT, also mediates the cholesterol efflux process of peripheral cells, and participates in the metabolism of many lipoproteins. Myeloperoxidase (MPO) and paroxypase 1 (Paraoxonase 1, PON1) are HDL affecting inflammation, and the associated protein.MPO of oxidative stress can selectively oxidize ApoA- I and weaken or lose the function of HDL. On the contrary, PON1 can directly participate in the hydrolysis of peroxide in lipoprotein to protect HDL from oxidation modification and protect the antioxidant function of HDL. The phosphatidylcholine acyl transferase (Lecithin cholesterol acyltransf erase, LCAT) can esterification the cholesterol, make the cholesterol enter into HDL, and make HDL gradually become rich in the HDL. The mature HDL.LCAT of cholesteryl ester is the key enzyme in the metabolism of HDL. When the function of LCAT is damaged, the synthesis of cholesteryl ester will be inhibited, which leads to hypercholesterolemia. At the same time, the maturation process of HDL will be blocked and the incidence of AS will increase. Loss of health, liver loss, deficiency of kidney essence, and kidney essence deficiency, external causes include diet, emotional disorder, and labor and leisure. On the pathogenesis of the disease, we think that the deficiency of this disease is true, with positive deficiency, wet turbid, phlegm coagulation, blood stasis as the standard, the three dirty function of spleen liver and kidney is the main basis of abnormal blood lipid. Kidney, invigorating the spleen and eliminating food, and activating blood and removing stasis, eliminating phlegm and collaterals as the treatment. It has been proved that many kinds of Chinese medicine monomers, single Chinese medicine and Chinese medicine compound can regulate blood lipid. Many reports suggest that Chinese medicine can improve the level of HDL, but at present, the research on lipid regulating Chinese medicine has not been influenced by the distribution of HDL subtypes to influence its function and has little influence. HDL anti atherosclerotic mechanism of anti atherosclerotic mechanism of traditional Chinese medicine was reported. In the previous experiment, we found that Xiong paehao capsule was effective in anti AS. The serum lipid TC, LDL increased with HDL in AS rabbits, and HDL had heterogeneity, and the elevation of HDL level was not necessarily on the basis of the early study of AS.. In this study, the subtypes, metabolism, components and functions of HDL were further studied. Objective: To explore the possible mechanism of Xiong Shao capsule against AS by setting up the rabbit atherosclerosis model and regulating the lipid metabolism and affecting the distribution and function of the HDL subtype. 1. groups and 60 male New Zealand rabbits were divided into two groups. The machine was divided into 5 groups: blank control group, model group, simvastatin group, Xiong Shao's low dose group, Xiong Shao's high dose group, each 12. The rabbit AS model was established by simple high fat feed feeding method. The method of administration: (1) the blank control group was fed with ordinary feed for 22 weeks; the model group was fed with high fat feed for 14 weeks and then fed with ordinary feed for 8 weeks; 3. The group was fed with high fat diet and simvastatin for 14 weeks, 8 weeks after feeding common feed and simvastatin, the dosage of simvastatin was 2mg/Kg D. 4. The low dose Xiong paehao group was fed with high fat feed and Chinese medicine for 14 weeks, and the ordinary feed and traditional Chinese medicine were fed for 8 weeks. The dose of Chinese medicine was Chuanxiong 1.5g/kg D and Radix Paeoniae 0.75g/kg. D. Feed and traditional Chinese medicine for 14 weeks, 8 weeks after feeding common feed and traditional Chinese medicine, the dose of traditional Chinese medicine was 3.0g/kg D, 1.5g/kg d.2. of Radix Paeoniae Rubra observed the formation of atherosclerotic plaques in the aorta after 22 weekend anesthesia, removed the thoracic aorta, and observed the formation of lipid plaque in the blood tube wall of the main specimens of the aorta. Neutral Faure Marin Young Liquid fixation, routine tissue paraffin section, hematoxylin eosin (HE) staining, microscopically observed histopathological changes of.3. high density lipoprotein and other blood lipids before the experiment, the drug was given 14 weeks, and the drug was given for 22 weeks and 3 time points. After centrifugation, the serum HDL and its component ApoA- I, total cholesterol (TC) were detected by the automatic biochemical analyzer. Low density lipoprotein (LDL), apolipoprotein B (ApoB), triglyceride (TG), extremely low density lipoprotein (VLDL) level.4. high density lipoprotein subtype, serum samples were detected by enzyme linked immunosorbent assay (ELISA), the level of HDL subtype HDL2 in serum was determined by.5. cholesterol reverse transport function test at the end of the experiment, and the liver tissue was frozen in liquid nitrogen. Izol method was used to extract total liver RNA, real-time fluorescent quantitative polymerase chain reaction (real-time PCR) for the determination of liver ABCAl mRNA, the expression of SR-BI mRNA, the antioxidant function of.6. high-density lipoprotein was detected by anisamine method, serum MPO activity was measured by anisamine method. Enzyme linked immunosorbent assay (ELISA) determination of the metabolism of serum PON1 activity high density lipoprotein metabolism The expression of LCAT mRNA in liver was measured by real time fluorescence quantitative polymerase chain reaction (real-time PCR). Results: 1. the aorta wall of rabbits in the blank control group was smooth, endothelial cells were continuous and no lipid deposition was found. The aorta wall of the rabbit model group was covered with lipid plaque, and a large amount of foam cells and smooth muscle layers were visible under the intima. A large amount of lipid deposition was found in the cells, and the lipid plaque on the aortic wall surface of the rabbits was less than that in the model group. Under the microscope, the foam cells of the intima were accumulated less.2. and 14 weeks. Except for the blank control group, the serum TC, TG, HDL, LDL, VLDL, ApoA-I, ApoB of the rabbits in each group were all increased, the difference was statistically significant (P0.05 or P0.01). After 22 weeks of administration, the serum TC, VLDL, and ApoA-I of the rabbits in the experimental group were higher than those in the blank control group. The difference was statistically significant (P0.05 or P0.01) in the model group (P0.05 or P0.01). The serum TC, TG, HDL, LDL, VLDL, ApoA-I, and ApoA-I were higher in the model group than in the blank control group for 14 weeks. There was no significant difference between the drug group and the model group. The serum TC, LDL, VLDL and ApoA-I in the model group were higher than that in the blank control group for 22 weeks. The difference was statistically significant (P0.01). The changes of serum HDL in the administration group were not significantly different from those in the model group; the serum TC, LDL, VLDL were lower in the administration group than the model group, and the ApoA-I was higher than the model group, and the difference was statistically significant. Meaning (P0.05 or P0.01), the serum TC, VLDL and ApoA-I were low in Xiong Shao, there was no significant difference between the high dose group and the simvastatin group for 22 weeks. The serum HDL2 in the model group was higher than that in the blank control group, the difference was statistically significant (P0.05), and the increase of serum HDL2 in the simvastatin group and Xiong Shao high dose group was larger than that in the model group, and the difference was statistically significant. The expression of ABCA1 mRNA in the model group was higher than that in the blank control group (P0.05). The expression of ABCA1 mRNA in the administration group was significantly higher than that in the model group (P0.01). The expression of SR-BI mRNA in the model group was significantly higher than that in the blank control group (P0.01). There was no significant difference in the expression of the SR-BI mRNA expression in the group of the drug groups compared with the model group (P0.05). The expression of ABCA1 mRNA in the model group was significantly higher than that in the model group (P0.01). After 14 weeks of administration, the activity of serum MPO in the model group was significantly higher than that in the blank control group (P0.01). The activity of MPO in each treatment group was lower than that in the model group (P0.05). After 22 weeks of administration, the activity of MPO in the model group was higher than that in the blank control group (P0.05), and the MPO in each group was lower than that in the model group (P0.05). The difference was statistically significant (P0.05). The changes of PON1 activity in rabbit serum were no difference (P0.05).6. administration at the end of 22 weeks. The expression of LCAT mRNA in the model group was significantly higher than that in the blank control group (P0.01). The expression of LCAT mRNA in each group was significantly higher than that in the model group (P0.01). Conclusion: 1. Xiong Shao Shao capsule can inhibit the formation of the aortic plaque in the aorta of AS rabbits, reduce the deposition of lipid on the inner wall of the blood vessels and reduce the bubbles. .2. Xiong Shao capsule can increase the level of ApoA-I in serum of AS rabbit and reduce the level of TC, LDL, VLDL. The regulation of lipid level.3. xiushao capsule can increase the level of HDL2 in the serum of AS rabbit, increase the level of HDL mature subtype, and influence the HDL subtype distribution of.4. Xiong Shao capsule to increase the expression of liver and promote the liver. Intracellular cholesterol metabolism, promote cholesterol reverse transport, its regulating effect on liver SR-BI mRNA is not obvious,.5. Xiong Shao capsule can reduce the activity of MPO in serum of AS rabbit, inhibit the ApoA-I in HDL, protect the antioxidant function of HDL, and its effect on serum PON1 activity is not obvious,.6. Xiong Shao capsule can up regulate AS rabbit liver LCAT The anti atherosclerosis mechanism of Xiong Shao capsule, the function of HDL, may be related to the mechanism of promoting the maturation of HDL, increasing the level of HDL mature subtype particles and regulating the level of blood lipid. The mechanism of anti atherosclerosis of Xiong paehao capsule of.8. may be related to increasing the expression of RCT related protein gene expression to promote RCT, and may protect the antioxidant function of HDL.
【学位授予单位】：北京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R285
本文编号：2159298
[Abstract]:Atherosclerosis is the pathological basis of various cardiovascular and cerebrovascular diseases. The pathogenesis of atherosclerosis is mainly lipid metabolism, inflammatory reaction, endothelial injury, oxidative stress, etc. High density lipoprotein (HDL) can pass cholesterol reverse transport (Reverse cholesterol transport, RCT), anti-inflammatory, antioxidant, Antithrombotic, anti apoptotic and diastolic blood vessels to protect blood vessels and anti atherosclerotic functions. The latest research has found that the level of HDL alone does not reflect its anti atherosclerotic function, and the function of HDL is closely related to the subtypes of subtypes, metabolism, and components of the.HDL, and the components and functions of the subtypes and functions are now becoming the present anti porridge. .ApoA- I, a hot spot in the study of sample sclerosis, is the main component of the protein and structure of HDL, and is also the main bearer of HDL to complete RCT, anti endothelial cell apoptosis, anti-oxidation, and anti-inflammatory function, the number of.ApoA- I decrease or structural change, the original function of HDL will be weakened or lost, and even the AS action.HDL via pre beta 1-HDL to HDL3 to HDL2. Gradually mature, HDL2, as a large mature HDL subtype, can promote the transport of cholesterol to the liver and the tissue of the synthetic steroid hormone, and the low HDL2 level is negatively related to the risk of coronary heart disease. The adenosine triphosphate binding cassette transporter A1 (ATP-binding cassette transporters A1, ABCA1) can be combined with HDL to promote intracellular cholesterol efflux. The effect of HDL particles formation, participation in RCT, regulating lipid metabolism. In addition, ABCA1 can also inhibit the expression of inflammatory factors and participate in the oxidative stress response in many ways, such as the AS process.B class type I scavenger receptor (Scavenger receptor class B, type I, SR-BI) can selectively absorb cholesterol esters in HDL, transmitted to liver and steroid irrigations The hormone producing tissue, completing RCT, also mediates the cholesterol efflux process of peripheral cells, and participates in the metabolism of many lipoproteins. Myeloperoxidase (MPO) and paroxypase 1 (Paraoxonase 1, PON1) are HDL affecting inflammation, and the associated protein.MPO of oxidative stress can selectively oxidize ApoA- I and weaken or lose the function of HDL. On the contrary, PON1 can directly participate in the hydrolysis of peroxide in lipoprotein to protect HDL from oxidation modification and protect the antioxidant function of HDL. The phosphatidylcholine acyl transferase (Lecithin cholesterol acyltransf erase, LCAT) can esterification the cholesterol, make the cholesterol enter into HDL, and make HDL gradually become rich in the HDL. The mature HDL.LCAT of cholesteryl ester is the key enzyme in the metabolism of HDL. When the function of LCAT is damaged, the synthesis of cholesteryl ester will be inhibited, which leads to hypercholesterolemia. At the same time, the maturation process of HDL will be blocked and the incidence of AS will increase. Loss of health, liver loss, deficiency of kidney essence, and kidney essence deficiency, external causes include diet, emotional disorder, and labor and leisure. On the pathogenesis of the disease, we think that the deficiency of this disease is true, with positive deficiency, wet turbid, phlegm coagulation, blood stasis as the standard, the three dirty function of spleen liver and kidney is the main basis of abnormal blood lipid. Kidney, invigorating the spleen and eliminating food, and activating blood and removing stasis, eliminating phlegm and collaterals as the treatment. It has been proved that many kinds of Chinese medicine monomers, single Chinese medicine and Chinese medicine compound can regulate blood lipid. Many reports suggest that Chinese medicine can improve the level of HDL, but at present, the research on lipid regulating Chinese medicine has not been influenced by the distribution of HDL subtypes to influence its function and has little influence. HDL anti atherosclerotic mechanism of anti atherosclerotic mechanism of traditional Chinese medicine was reported. In the previous experiment, we found that Xiong paehao capsule was effective in anti AS. The serum lipid TC, LDL increased with HDL in AS rabbits, and HDL had heterogeneity, and the elevation of HDL level was not necessarily on the basis of the early study of AS.. In this study, the subtypes, metabolism, components and functions of HDL were further studied. Objective: To explore the possible mechanism of Xiong Shao capsule against AS by setting up the rabbit atherosclerosis model and regulating the lipid metabolism and affecting the distribution and function of the HDL subtype. 1. groups and 60 male New Zealand rabbits were divided into two groups. The machine was divided into 5 groups: blank control group, model group, simvastatin group, Xiong Shao's low dose group, Xiong Shao's high dose group, each 12. The rabbit AS model was established by simple high fat feed feeding method. The method of administration: (1) the blank control group was fed with ordinary feed for 22 weeks; the model group was fed with high fat feed for 14 weeks and then fed with ordinary feed for 8 weeks; 3. The group was fed with high fat diet and simvastatin for 14 weeks, 8 weeks after feeding common feed and simvastatin, the dosage of simvastatin was 2mg/Kg D. 4. The low dose Xiong paehao group was fed with high fat feed and Chinese medicine for 14 weeks, and the ordinary feed and traditional Chinese medicine were fed for 8 weeks. The dose of Chinese medicine was Chuanxiong 1.5g/kg D and Radix Paeoniae 0.75g/kg. D. Feed and traditional Chinese medicine for 14 weeks, 8 weeks after feeding common feed and traditional Chinese medicine, the dose of traditional Chinese medicine was 3.0g/kg D, 1.5g/kg d.2. of Radix Paeoniae Rubra observed the formation of atherosclerotic plaques in the aorta after 22 weekend anesthesia, removed the thoracic aorta, and observed the formation of lipid plaque in the blood tube wall of the main specimens of the aorta. Neutral Faure Marin Young Liquid fixation, routine tissue paraffin section, hematoxylin eosin (HE) staining, microscopically observed histopathological changes of.3. high density lipoprotein and other blood lipids before the experiment, the drug was given 14 weeks, and the drug was given for 22 weeks and 3 time points. After centrifugation, the serum HDL and its component ApoA- I, total cholesterol (TC) were detected by the automatic biochemical analyzer. Low density lipoprotein (LDL), apolipoprotein B (ApoB), triglyceride (TG), extremely low density lipoprotein (VLDL) level.4. high density lipoprotein subtype, serum samples were detected by enzyme linked immunosorbent assay (ELISA), the level of HDL subtype HDL2 in serum was determined by.5. cholesterol reverse transport function test at the end of the experiment, and the liver tissue was frozen in liquid nitrogen. Izol method was used to extract total liver RNA, real-time fluorescent quantitative polymerase chain reaction (real-time PCR) for the determination of liver ABCAl mRNA, the expression of SR-BI mRNA, the antioxidant function of.6. high-density lipoprotein was detected by anisamine method, serum MPO activity was measured by anisamine method. Enzyme linked immunosorbent assay (ELISA) determination of the metabolism of serum PON1 activity high density lipoprotein metabolism The expression of LCAT mRNA in liver was measured by real time fluorescence quantitative polymerase chain reaction (real-time PCR). Results: 1. the aorta wall of rabbits in the blank control group was smooth, endothelial cells were continuous and no lipid deposition was found. The aorta wall of the rabbit model group was covered with lipid plaque, and a large amount of foam cells and smooth muscle layers were visible under the intima. A large amount of lipid deposition was found in the cells, and the lipid plaque on the aortic wall surface of the rabbits was less than that in the model group. Under the microscope, the foam cells of the intima were accumulated less.2. and 14 weeks. Except for the blank control group, the serum TC, TG, HDL, LDL, VLDL, ApoA-I, ApoB of the rabbits in each group were all increased, the difference was statistically significant (P0.05 or P0.01). After 22 weeks of administration, the serum TC, VLDL, and ApoA-I of the rabbits in the experimental group were higher than those in the blank control group. The difference was statistically significant (P0.05 or P0.01) in the model group (P0.05 or P0.01). The serum TC, TG, HDL, LDL, VLDL, ApoA-I, and ApoA-I were higher in the model group than in the blank control group for 14 weeks. There was no significant difference between the drug group and the model group. The serum TC, LDL, VLDL and ApoA-I in the model group were higher than that in the blank control group for 22 weeks. The difference was statistically significant (P0.01). The changes of serum HDL in the administration group were not significantly different from those in the model group; the serum TC, LDL, VLDL were lower in the administration group than the model group, and the ApoA-I was higher than the model group, and the difference was statistically significant. Meaning (P0.05 or P0.01), the serum TC, VLDL and ApoA-I were low in Xiong Shao, there was no significant difference between the high dose group and the simvastatin group for 22 weeks. The serum HDL2 in the model group was higher than that in the blank control group, the difference was statistically significant (P0.05), and the increase of serum HDL2 in the simvastatin group and Xiong Shao high dose group was larger than that in the model group, and the difference was statistically significant. The expression of ABCA1 mRNA in the model group was higher than that in the blank control group (P0.05). The expression of ABCA1 mRNA in the administration group was significantly higher than that in the model group (P0.01). The expression of SR-BI mRNA in the model group was significantly higher than that in the blank control group (P0.01). There was no significant difference in the expression of the SR-BI mRNA expression in the group of the drug groups compared with the model group (P0.05). The expression of ABCA1 mRNA in the model group was significantly higher than that in the model group (P0.01). After 14 weeks of administration, the activity of serum MPO in the model group was significantly higher than that in the blank control group (P0.01). The activity of MPO in each treatment group was lower than that in the model group (P0.05). After 22 weeks of administration, the activity of MPO in the model group was higher than that in the blank control group (P0.05), and the MPO in each group was lower than that in the model group (P0.05). The difference was statistically significant (P0.05). The changes of PON1 activity in rabbit serum were no difference (P0.05).6. administration at the end of 22 weeks. The expression of LCAT mRNA in the model group was significantly higher than that in the blank control group (P0.01). The expression of LCAT mRNA in each group was significantly higher than that in the model group (P0.01). Conclusion: 1. Xiong Shao Shao capsule can inhibit the formation of the aortic plaque in the aorta of AS rabbits, reduce the deposition of lipid on the inner wall of the blood vessels and reduce the bubbles. .2. Xiong Shao capsule can increase the level of ApoA-I in serum of AS rabbit and reduce the level of TC, LDL, VLDL. The regulation of lipid level.3. xiushao capsule can increase the level of HDL2 in the serum of AS rabbit, increase the level of HDL mature subtype, and influence the HDL subtype distribution of.4. Xiong Shao capsule to increase the expression of liver and promote the liver. Intracellular cholesterol metabolism, promote cholesterol reverse transport, its regulating effect on liver SR-BI mRNA is not obvious,.5. Xiong Shao capsule can reduce the activity of MPO in serum of AS rabbit, inhibit the ApoA-I in HDL, protect the antioxidant function of HDL, and its effect on serum PON1 activity is not obvious,.6. Xiong Shao capsule can up regulate AS rabbit liver LCAT The anti atherosclerosis mechanism of Xiong Shao capsule, the function of HDL, may be related to the mechanism of promoting the maturation of HDL, increasing the level of HDL mature subtype particles and regulating the level of blood lipid. The mechanism of anti atherosclerosis of Xiong paehao capsule of.8. may be related to increasing the expression of RCT related protein gene expression to promote RCT, and may protect the antioxidant function of HDL.
【学位授予单位】：北京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R285
【参考文献】
相关期刊论文 前10条
1 谢宗恺;高蔚娜;蒲玲玲;韦京豫;张敏;郭长江;;高脂培养条件下槲皮素对大鼠肝细胞胆固醇代谢的影响[J];营养学报;2016年01期
2 王晓娜;叶平;;高密度脂蛋白胆固醇与动脉粥样硬化研究进展[J];中华保健医学杂志;2015年06期
3 商亮;曹佳;喻红;;对氧磷酶1在高密度脂蛋白结构与功能中的意义[J];中国动脉硬化杂志;2015年12期
4 秦树存;于杨;赵亚南;吴云;;磷脂转运蛋白在高密度脂蛋白代谢和胆固醇逆向转运中的生物学作用[J];中国动脉硬化杂志;2015年12期
5 罗伟;马建伟;;原发性血脂异常患者中医体质类型与中医证型相关性研究[J];解放军医药杂志;2015年09期
6 林小龙;温东辉;罗浩元;朱丽明;曾涛;邓宇伟;;大蒜素通过上调ABCA1表达减少脂质在THP1巨噬细胞源性泡沫细胞中蓄积[J];中国生物化学与分子生物学报;2015年09期
7 胡怀强;王兴臣;周永红;;从血浊论治动脉粥样硬化的理论探析[J];中国中医药现代远程教育;2015年11期
8 赵慧;刘红;李一石;;高密度脂蛋白组分:心血管疾病诊治的新靶点[J];心血管病学进展;2015年03期
9 吴厚平;胡允兆;;氧化型低密度脂蛋白与冠心病的相关性研究进展[J];中国医学创新;2015年14期
10 韩耀霞;张强;梁斌;张娜娜;边云飞;肖传实;;ABCA1在冠心病及其发病机制中的研究新进展[J];中国动脉硬化杂志;2015年04期
相关博士学位论文 前3条
1 张艳虹;芎芍胶囊对动脉粥样硬化兔胆固醇逆向转运及炎症反应的影响[D];北京中医药大学;2014年
2 宋庆桥;冠心病心绞痛患者中医证候与相关血清蛋白表达的研究[D];中国中医科学院;2010年
3 孙晓伟;不稳定性心绞痛方证对应及证候动态变化研究[D];北京中医药大学;2010年
相关硕士学位论文 前9条
1 盛松;芎芍胶囊对动脉粥样硬化兔血脂及炎症因子的影响[D];北京中医药大学;2013年
2 李佳;血脉宁颗粒对高脂饮食家兔血清oxLDL、ABCA1含量影响的实验研究[D];辽宁中医药大学;2013年
3 刘倩;白细胞介素-8对动脉粥样硬化斑块稳定性的影响及芎芍胶囊的干预效应研究[D];中国中医科学院;2012年
4 钟建开;普罗布考在动脉粥样硬化模型中改善高密度脂蛋白功能的研究[D];南方医科大学;2011年
5 李宝石;补心通脉颗粒对动脉粥样硬化家兔氧化应激的影响[D];南方医科大学;2011年
6 范永臻;过氧化酶体增殖物激活型受体γ激动剂对动脉粥样硬化兔HDL功能影响及其调控机制探讨[D];南方医科大学;2010年
7 王振坤;普罗布考对动脉粥样硬化兔HDL功能的影响及其调控机制探讨[D];南方医科大学;2010年
8 庞文艳;芪参益气滴丸对冠心病病人高密度脂蛋白亚型和炎性因子的影响[D];天津医科大学;2009年
9 汪韶君;扇贝裙边糖胺聚糖对巨噬细胞脂蛋白代谢及脂蛋白受体表达的影响研究[D];青岛大学;2006年
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