全反式维甲酸对动脉粥样硬化兔内皮功能调节的探究

发布时间：2018-05-04 07:47

本文选题：ATRA + 动脉粥样硬化　；参考：《安徽医科大学》2017年硕士论文

【摘要】：背景众所周知,动脉粥样硬化是一种发生在动脉内壁的慢性炎症性疾病,其发病机制尚未完全阐明。越来越多的证据表明,血管内皮功能障碍与心血管事件密切相关。因此,正确认识影响血管的收缩和舒张功能的因子失衡,对于了解并干预动脉粥样硬化的发病起重要作用。内皮细胞可以分泌调节因子,包括一氧化氮(NO)和内皮素(ET),从而调节血管松弛和收缩。全反式维甲酸(Retinoic Acid,ATRA)是维生素A的天然衍生物,可通过调节炎症因子来改善动脉粥样硬化。然而,关于全反式维甲酸对动脉内皮功能的作用的研究较少见。本文,我们通过研究在动脉粥样硬化兔中,全反式维甲酸对血管内皮产生的内皮素-1(ET-1)和一氧化氮(NO)水平的影响来探讨全反式维甲酸对动脉粥样硬化的作用机制。目的本研究选择普通新西兰白兔作为动物模型,观察全反式维甲酸对动脉粥样硬化兔血管内皮的内皮素-1和一氧化氮的影响,从而探讨全反式维甲酸对动脉粥样硬化的作用机制。方法24只普通新西兰白兔适应性饲养一周后,随机分为三组(n=8):A为正常对照组,B为高脂组,C为ATRA干预组。正常对照组予以普通饲料喂养,高脂组给予高脂饲料喂养,ATRA干预组为高脂饲料喂养,高脂饮食4周后,开始加ATRA(10mg/(kg·d))灌胃干预,并继续予以高脂饮食喂养。12周后,采集血标本和胸主动脉标本,观察主动脉斑块形成情况,测定兔的血脂水平、动脉内膜通透性、血液e NOS活性、内皮素(ET-1)、血浆一氧化氮(NO)含量;采用Western blot检测主动脉中ET-1,内皮一氧化氮合酶(e NOS)和e NOS磷酸化的产物(p-e NOS)的水平。结果1.ATRA减轻动脉粥样硬化兔血管内壁斑块形成:油红O染色肉眼可见,正常对照组,血管内壁光滑,基本无斑块形成。高脂组内壁可见明显大面积斑块形成,凹凸不平。ATRA干预组主动脉内壁有斑块形成,与高脂组相比明显较少。HE染色后,光镜下可见正常组动脉壁内膜光滑连续,无内膜增厚,平滑肌细胞排列整齐,弹性纤维稍紊乱。与正常组相比,高脂组动脉壁内膜显著增厚,弹力板多处断裂,可见粥样斑块大量形成,斑块中可见炎性细胞、泡沫细胞。ATRA干预组在药物干预后,其动脉壁内膜可见少量斑块形成,与高脂组相比,脂肪细胞和炎性细胞明显减少。2.ATRA降低动脉粥样硬化兔的血脂水平:由于脂质水平与动脉粥样硬化的进程关系密切,我们检测了各组的胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白。与正常组相比较,高脂组的胆固醇、甘油三酯、低密度脂蛋白明显增高(P0.05),高密度脂蛋白降低(P0.05)。而ATRA干预组的胆固醇、甘油三酯、低密度脂蛋白均比高脂组减低,高密度脂蛋白较高(P0.05)。3.ATRA降低动脉粥样硬化兔的动脉内皮通透性:在荧光显微镜下,可以观察到荧光染料在对照组中几乎没有穿透,但渗透高脂组动脉壁的全层,ATRA干预组中荧光染料的穿透率显着低于高脂组,但比对照组增加。4.ATRA影响动脉粥样硬化兔主动脉中ET-1,e NOS和p-e NOS的表达:Western blot结果显示,与对照组相比较,高脂组兔动脉组织中ET-1的表达明显升高,而ATRA干预组的ET-1表达量介于对照组和高脂组之间。此外,关于e NOS磷酸化的产物Ser-1177(p-e NOS)的水平,p-e NOS在高脂组中的表达低于对照组,在用ATRA干预之后,其表达量明显增加。而ATRA的干预对e NOS的表达并无统计学意义。5.ATRA对动脉粥样硬化兔血中e NOS活性和ET-1、NO水平的影响:结果表明,高脂组NO含量显着低于对照组(P0.05),ATRA处理后eNOS活性和NO含量显着升高(P0.05)。ET-1的检测结果与NO的水平相反,ATRA组的ET-1表达明显低于高脂组(P0.05)。结论NO和ET之间的平衡调节是预防和治疗动脉粥样硬化的一个重要途径。在本实验中显示,ATRA可以减少血浆中ET-1的含量,通过增加e NOS磷酸化增加NO含量,并通过调节两者之间的平衡,从而改善血管松弛,减轻动脉粥样硬化斑块形成。它将为未来动脉粥样硬化的临床治疗提供新的方向。
[Abstract]:As we all know, atherosclerosis is a chronic inflammatory disease occurring on the inner wall of the artery. Its pathogenesis has not been fully elucidated. More and more evidence suggests that vascular endothelial dysfunction is closely related to cardiovascular events. Therefore, it is necessary to understand the imbalance of factors affecting vasodilation and diastolic function of blood vessels and to understand and do it. Preatheromatous disease plays an important role. Endothelial cells secrete regulatory factors, including nitric oxide (NO) and endothelin (ET), which regulate vascular relaxation and contraction. Retinoic Acid (ATRA) is a natural derivative of vitamin A, which can be used to improve atherosclerosis by regulating inflammatory factors. The effect of trans retinoic acid on the function of arterial endothelial cells is rare. In this article, we studied the effect of all trans retinoic acid on the levels of endothelin -1 (ET-1) and nitric oxide (NO) produced by vascular endothelium in atherosclerotic rabbits. The effect of all trans retinoic acid on endothelin -1 and nitric oxide in vascular endothelial cells of atherosclerotic rabbits was observed as an animal model, and the mechanism of all trans retinoic acid on atherosclerosis was investigated. Methods 24 New Zealand rabbits were randomly divided into three groups (n=8) after a week of adaptive feeding: A was normal. Group B was high fat group and C was ATRA intervention group. Normal control group was fed with ordinary diet, high fat group was fed with high fat diet, ATRA intervention group was fed with high fat diet, and after 4 weeks of high fat diet, ATRA (10mg/ (kg. D)) was added to stomach intervention, and the high fat diet was continued to be fed for.12 weeks, and blood specimens and thoracic aorta specimens were collected to observe the main body of the aorta. Atherosclerotic plaque formation, blood lipid level, intima permeability, e NOS activity, endothelin (ET-1), plasma nitric oxide (NO) levels, and the levels of ET-1, endothelial nitric oxide synthase (E NOS) and E NOS phosphorylation in the aorta of the aorta were measured by Western blot. Plaque formation in the inner wall: oil red O staining was visible to the naked eye, in the normal control group, the inner wall of the blood vessel was smooth, and the plaque formed basically. The inner wall of the high fat group showed obvious large area plaque formation. The inner wall of the aorta was formed in the unflat.ATRA intervention group. The smooth connection of the intima of the normal artery wall was visible in the normal group after the light microscope compared with the high fat group. Continued, without intimal thickening, smooth muscle cells arranged neatly, elastic fiber slightly disorder. Compared with the normal group, the intima of the arterial wall of the high fat group was thickened, the elastic plate was fractured in many places, the atherosclerotic plaque was seen in a large number, the plaque was visible in the plaque, and the.ATRA intervention group of the foam cells showed a small amount of plaque formation in the intima of the arterial wall. Compared with the high fat group, the fat cells and inflammatory cells significantly reduced.2.ATRA to reduce the level of blood lipid in atherosclerotic rabbits. Due to the close relationship between the lipid level and the process of atherosclerosis, we detected the cholesterol, triglycerides, high density lipoprotein and low density lipoprotein in each group. Oil three ester, low density lipoprotein (P0.05) and high density lipoprotein decreased (P0.05). The cholesterol, triglyceride and low density lipoprotein in the ATRA intervention group were lower than those in the high fat group, and the high density lipoprotein (P0.05).3.ATRA reduced the arterial endothelial permeability in atherosclerotic rabbits: fluorescence staining could be observed under the fluorescence microscope. There was little penetration in the control group, but the penetration rate of the fluorescent dye in the ATRA intervention group was significantly lower than that in the high fat group, but the increase of.4.ATRA in the aorta of the atherosclerotic rabbits influenced the expression of ET-1, e NOS and P-E NOS in the atherosclerotic rabbits. The result of Western blot showed that the rabbit artery in the high fat group was compared with the control group. The expression of ET-1 in the tissue was significantly increased, while the expression of ET-1 in the ATRA intervention group was between the control group and the high fat group. In addition, the expression of P-E NOS in the high fat group was lower than the control group on the level of the product Ser-1177 (P-E NOS) of the product of E NOS phosphorylation. The effect of statistical significance.5.ATRA on the activity of E NOS and the level of ET-1 and NO in the blood of atherosclerotic rabbits showed that the content of NO in the high fat group was significantly lower than that of the control group (P0.05), and the eNOS activity and the NO content of the NO increased significantly (P0.05) after ATRA treatment (P0.05), and the expression of.ET-1 was significantly lower than that of the high fat group. Balance regulation with ET is an important way to prevent and treat atherosclerosis. In this experiment, ATRA can reduce the content of ET-1 in plasma, increase NO content by increasing e NOS phosphorylation, and regulate the balance between the two, thus improving vascular relaxation and reducing the formation of atherosclerotic plaques. To provide new directions for clinical treatment of atherosclerosis.

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R543.5

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