吸烟冠心病患者HDL对THP细胞胆固醇逆转运的影响及机制

发布时间：2018-04-10 06:27

  本文选题：胆固醇逆转运　切入点：动脉粥样硬化　出处：《山西医科大学》2017年硕士论文

【摘要】：目的:本研究通过提取正常健康人及吸烟冠心病患者的HDL培养THP-1细胞源性泡沫细胞,观察吸烟冠心病患者的HDL对其胆固醇逆转运的影响及机制,为吸烟促进动脉粥样硬化发病机制提供理论基础。方法:采集正常健康人群及吸烟冠心病人群血样,利用超高速离心机提取高密度脂蛋白(HDL);利用佛波酯诱导THP-1细胞成为巨噬细胞,然后利用氧化低密度脂蛋白(ox-LDL)诱导上述细胞形成泡沫细胞,并将其分为3组,分别为:空白对照组、正常人HDL组、吸烟冠心病HDL组。利用油红O染色观察分组泡沫细胞内脂滴情况;利用液体闪烁计数仪从表观现象方面检测各组细胞相应的胆固醇流出率;采用Western blot检测以上3组三磷酸腺苷结合盒转运体A1(ABCA1)、三磷酸腺苷结合转运体G1(ABCG1)、B类Ⅰ型清道夫受体(SR-BI)相应蛋白质含量。结果:⑴成功提取人HDL,并鉴定;⑵佛波酯成功诱导THP-1细胞形成泡沫细胞,ox-LDL成功诱导泡沫细胞形成;⑶和空白对照组相比,正常人HDL及吸烟冠心病HDL组均可使胆固醇流出率增加(P0.05),正常人HDL组作用较吸烟冠心病HDL组作用显著(P0.05);⑷与空白对照组相比,正常人HDL组ABCA1、ABCG1蛋白表达无明显变化,吸烟冠心病HDL组ABCA1、ABCG1蛋白表达显著减少(P0.05);⑸与空白对照组相比,正常人HDL组及吸烟冠心病HDL组SR-B1蛋白表达显著升高,正常人HDL组作用较吸烟冠心病HDL组作用显著(P0.05)。结论:正常人HDL可以促进THP-1细胞源性泡沫细胞胆固醇逆转运。其可以促进胆固醇流出,且可以升高SR-B1蛋白表达,但是其并不影响ABCA1、ABCG1的蛋白表达量,而吸烟冠心病人HDL相比正常人HDL,促进胆固醇流出作用及升高SR-B1蛋白表达作用减弱,且可以显著减低ABCA1、ABCG1蛋白表达量,为吸烟促进动AS发病机制提供理论基础,从而减弱或者避免AS发生和发展,为冠状动脉粥样硬化性心脏病的临床预防和治疗提供理论依据。
[Abstract]:Objective: to investigate the effect of HDL on cholesterol reverse transport and its mechanism by extracting HDL from healthy subjects and smoking patients with coronary heart disease (CHD) to culture THP-1 cell-derived foam cells.To provide a theoretical basis for smoking to promote the pathogenesis of atherosclerosis.Methods: the blood samples of healthy and smoking patients with coronary heart disease were collected, high density lipoprotein (HDL) was extracted by ultrahigh speed centrifuge, and THP-1 cells were induced into macrophages by phorbol ester.Then the cells were induced to form foam cells by oxidized low density lipoprotein (ox-LDL) and divided into three groups: blank control group, normal HDL group, smoking coronary heart disease HDL group.Lipid droplets in foam cells were observed by oil red O staining and cholesterol efflux was detected by liquid scintillation counter.The protein contents of adenosine triphosphate binding cassette transporter A _ (1) ABCA _ (1) and adenosine triphosphate binding transporter G _ (1) ABCG _ (1) B type 鈪，

本文编号：1730078