健康个体与冠心病患者外周血单核源性巨噬细胞泡沫化过程的比较  

Transformation of human peripheral blood monocyte-derived macrophages to foam cells in healthy and coronary heart disease cases

作　　者：曹保卫 吴鹏[2] 覃学美 彭怡兰 梁东辉[3] 

机构地区：[1]来宾市人民医院心血管内科,广西壮族自治区来宾市546100 [2]南方医科大学附属珠江医院心血管内科,广东省广州市510280 [3]南方医科大学附属珠江医院中医科,广东省广州市510280

出　　处：《中国组织工程研究与临床康复》2008年第28期5558-5561,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省自然科学基金资助项目(06024444)~~

摘　　要：背景:不同疾病状态的个体由于体内生化病理环境的差异,其单核-巨噬等细胞对各种体内外刺激因素的反应性亦存在一定差异。目的:拟观察健康个体与冠心病患者外周血单核源性巨噬细胞泡沫化过程的差异。设计、时间及地点:细胞水平的对比观察实验,于2007-06/12在珠江医院中心实验室完成。对象:选择本院收治的冠心病高危及冠心病患者,纳入标准:符合冠心病的临床诊断标准。另选健康个体作为对照,均经临床体格检查和生化检查符合健康人判断标准,受试者对试验知情同意。方法:采用一次性密度梯度超速离心法从血浆中分离低密度脂蛋白,以Cu2+诱导法对其进行氧化修饰。分别采用密度梯度离心法和塑料吸附法从健康个体、冠心病高危及冠心病患者外周血中分离单核细胞,以80nmol/L佛波酯刺激48h使之转化为巨噬细胞。实验按单核细胞取材群体不同分为3组,即健康个体组、冠心病高危组和冠心病组,同时设对照组,对照组只加培养基,实验组另加氧化低密度脂蛋白至终末浓度为80mg/L。主要观察指标:培养24h和48h后收集细胞,油红O染色观察细胞内脂滴变化;高效液相色谱分析细胞内胆固醇的变化。结果:单核细胞经80nmol/L佛波酯诱导24h后,光学显微镜下可见冠心病高危组和冠心病组大部分细胞已由圆形变成多角形状,形态变化较对照组及健康个体组明显。48h后,光镜下可见单核细胞已转化为典型的巨噬细胞。油红O染色发现,与80mg/L氧化低密度脂蛋白共孵育48h后培养细胞中出现了大量的红染颗粒。高效液相色谱法测定细胞内胆固醇提示细胞内总胆固醇明显增加,其中胆固醇酯含量大于游离胆固醇,符合泡沫细胞的特征。同时发现,来自冠心病患者体内的单核细胞更易被诱导形成巨噬细胞,且其发生泡沫化程度较健康、冠心病高危个体者高。结论:源自冠心病高危�BACKGROUND： Monocyte-derived macrophage responses to in vivo and vitro stimulating factors vary to different in vivo biochemical environments in individuals with varying diseases. OBJECTIVE： To compare the difference of the transformation of human peripheral blood monocyte-derived macrophages to foam cells in healthy and coronary heart disease （CHD） cases. DESIGN, TIME AND SETTING： Comparative observation at cell level was performed at the Central Laboratory of Zhujiang Hospital from June to December 2007. PARTICIPANTS： CHD high-risk and CHD patients were selected according to the clinically diagnostic criteria of CHD. In addition, healthy people that were confirmed by clinical physical and biochemical examinations were selected. The informed consent was obtained from all subjects. METHODS： Low density lipoprotein （LDL） was isolated from human plasma by one time density gradient centrifugation, and Cu^2＋ was used to induce the oxidation of LDL （ox-LDL）. Monocytes were isolated respectively from peripheral blood of healthy cases, CHD high-risk cases and CHD cases by Ficoll-Hypaque density gradient centrifugation and plastic adsorptive process. The isolated cells were induced and stimulated by phorbol 12-myristate 13-acetate （PMA） at concentration of 80 nmol/L for 48 h so as to make them transfer to macrophages. The successfully transferred cells were then co-incubated with ox-LDL at concentration of 80 mg/L for 48 hours. There were 4 groups： healthy group, CHD high-risk group, CHD group, and control group （only medium）. MAIN OUTCOME MEASURES： Oil red O staining technique was used to identify the formation of foam cells; cholesterol changes were detected by high performance liquid chromatography （HPLC） and Modified Lowry protein assay after 24 and 48 hours of culture. RESULTS： The isolated monocytes transferred from round to polygon-shaped in CHD high-risk group and CHD group macrophages after inducing by 80 nmol/L PMA for 48 hours, significantly different from

关 键 词：冠状动脉粥样硬化性心脏病 单核细胞 巨噬细胞 泡沫细胞 

分 类 号：R331[医药卫生—人体生理学]