3-羟基苯甲醛抑制兔自体移植静脉桥内膜增生的研究

发布时间：2018-04-10 16:14

本文选题：3-羟基苯甲醛 + 静脉桥　；参考：《广西医科大学》2017年硕士论文

【摘要】：研究背景及目的心血管疾病是最常见的疾病病种,冠心病是心血管疾病重要的组成部分,也是心血管疾病中最常见的死因。对于严重的冠心病,冠状动脉旁路移植术是其主要的治疗方法。外科手术常用的取材有两种:左乳内动脉和大隐静脉。相较于乳内动脉血管桥的较好的远期通畅率,大隐静脉血管桥远期通畅率不容乐观。3-羟基苯甲醛(3-HBA)是一种原儿茶醛的前体,有研究表明原儿茶醛类物质有心血管保护作用。但并无3-HBA对静脉桥作用的研究。本实验通过研究3-HBA对人大隐静脉血管平滑肌细胞及兔自体移植静脉桥内膜增生的影响,以其寻找一种新的能够防治冠状动脉旁路移植术后静脉桥狭窄的药物。研究方法1.人大隐静脉平滑肌细胞进行体外培养,设立对照组,以及药物梯度组(25,50,100μmol/L),作用人大隐静脉平滑肌细胞24h后,用MTT法检测细胞增殖情况,Transwell小室法检测细胞迁移情况,流式细胞仪(Flow Cytometry,FCM)检测细胞凋亡情况,Western Blot检测p-AKT蛋白表达情况。2.选取24只健康的实验用新西兰兔,不限雌雄,体重在2000g-2500g之间,编号,计算机随机法分成实验组A组和对照组B组,每组再均分为两个亚组,即A1、A2组和B1、B2组,每个亚组6只兔子,建立同侧颈外静脉移植到颈总动脉的实验动物模型,A组动物从耳缘静脉注射3-HBA 100mg/(kgd),B组动物给予同剂量生理盐水以作对照。给药时间一直持续至取材前一天。A1、B1组动物于模型建立后的第2周采集移植静脉桥,A2、B2组动物于模型建立后4周采集移植静脉桥,计算机图形软件计算静脉桥内膜厚度,并用蛋白印记法(Western Blot)对各组移植静脉桥中AKT蛋白的磷酸化情况进行检测。结果1.药物干预人大隐静脉平滑肌细胞24h后,相比于对照组,实验组细胞增殖与迁移能力均受到明显抑制,并呈现一定的剂量依赖性,流式细胞仪凋亡检测发现其中晚期凋亡细胞比例明显增加,Western blot实验发现p-AKT表达明显减少,p0.05,差异均有统计学意义。2.24只兔子行搭桥模型成功,无明显的手术切口感染。术后2周时,实验组A1组静脉桥内膜厚度为76.57±6.76μm,而同期对照组B1组静脉桥内膜厚度为87.32±6.89μm,实验组组静脉桥内膜厚度较对照组已经稍有减小,p=0.123,差异无统计学意义;到4周时,实验组A2内膜厚度为45.39±9.41μm,对照组B2组内膜厚度为134.07±18.80μm,p0.05,差异具有统计学意义。无论2周或者4周时,p-AKT的表达量实验组组均较对照组均明显降低,p0.05,差异有统计意义。结论3-羟基苯甲醛能抑制实验用新西兰兔自体移植静脉桥内膜增生,可能与其抑制AKT的磷酸化从而抑制平滑肌细胞增殖迁移以及促进VSMC凋亡有关。
[Abstract]:Background and objective Cardiovascular disease is the most common disease. Coronary heart disease is an important part of cardiovascular disease and the most common cause of death in cardiovascular disease.Coronary artery bypass grafting is the main treatment for severe coronary heart disease.There are two commonly used materials for surgery: the left internal mammary artery and the great saphenous vein.But there was no study of the effect of 3-HBA on venous bridge.The aim of this study was to investigate the effects of 3-HBA on intimal hyperplasia of vascular smooth muscle cells of human saphenous vein and autograft vein graft in rabbits in order to find a new drug to prevent and treat the stenosis of vein graft after coronary artery bypass grafting (CABG).Method 1.Human saphenous vein smooth muscle cells were cultured in vitro. Control group and drug gradient group (2550100 渭 mol / L) were used to detect cell proliferation and cell migration by MTT assay.Flow Cytometry (FCM) was used to detect apoptosis. Western Blot was used to detect the expression of p-AKT protein.Twenty-four healthy New Zealand rabbits were randomly divided into experimental group A and control group B, each group was divided into two subgroups: A1A 2 group and B 1 B 2 group, with 6 rabbits in each subgroup.An experimental animal model of ipsilateral external jugular vein transplantation to the common carotid artery was established. The animals of group A were injected with 3-HBA 100mg / kg / kg 3-HBA from the auricular margin to control the rats in group B with the same dose of normal saline.The administration time lasted until the first day before the drug was taken. Group A _ (1) B _ (1) collected the grafted vein grafts 4 weeks after the establishment of the model, and the thickness of the vein grafts was calculated by computer graphics software, and the thickness of the vein grafts was calculated by computer graphics software in the second week after the establishment of the model.The phosphorylation of AKT protein in vein grafts was detected by Western blot.Result 1.Compared with the control group, the proliferation and migration ability of the experimental group were significantly inhibited after 24 hours of drug intervention, and in a dose-dependent manner.The proportion of late apoptotic cells was significantly increased by flow cytometry. Western blot assay showed that the expression of p-AKT decreased significantly (P 0.05), and the difference was statistically significant. 2.24 rabbits were successful in bypass grafting model, and there was no obvious wound infection.At 2 weeks after operation, the intimal thickness of venous grafts was 76.57 卤6.76 渭 m in group A1 and 87.32 卤6.89 渭 m in group B1, respectively.The intimal thickness of experimental group A2 was 45.39 卤9.41 渭 m, and that of control group B2 was 134.07 卤18.80 渭 m (p0.05).The expression of p-AKT in the experimental group was significantly lower than that in the control group at 2 or 4 weeks, and the difference was statistically significant.Conclusion 3-hydroxybenzaldehyde can inhibit the proliferation of autografted vein graft intima in New Zealand rabbits, which may be related to the inhibition of AKT phosphorylation, the inhibition of smooth muscle cell proliferation and migration, and the promotion of VSMC apoptosis.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R654.2

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