ADMA对人THP-1源巨噬细胞和泡沫细胞内胆固醇含量影响的研究
发布时间：2019-04-02 01:44
【摘要】： 目的ADMA是一种强烈的内源性一氧化氮合酶(NOS)抑制剂,被认为是心血管疾病的一个新的危险因子。单核细胞／巨噬细胞及其形成的泡沫细胞在动脉粥样硬化的发生发展中具有重要作用。本研究探讨ADMA对人THP-1源巨噬细胞和泡沫细胞内胆固醇含量的影响。 方法体外培养人THP-1单核细胞,给予佛波酯(PMA)诱导分化成巨噬细胞,用免疫组化法鉴定THP-1源巨噬细胞；THP-1源巨噬细胞给予氧化低密度脂蛋白(ox-LDL)诱导分化为THP-1源泡沫细胞,用油红O染色鉴定THP-1源泡沫细胞。用酶偶联比色法检测不同浓度ADMA(3.75、7.5、15、30μmol/L)作用不同时间(6、12、24h)THP-1源巨噬／泡沫细胞内胆固醇含量。 结果免疫组化结果表明佛波酯诱导后85%THP-1单核细胞表达CD68，THP-1细胞被诱导为THP-1源巨噬细胞；油红O染色结果显示细胞内有大量脂滴存在,THP-1源巨噬细胞吞噬ox-LDL形成THP-1源泡沫细胞。酶偶联比色法结果显示ADMA在体外以浓度-时间依赖的方式增加THP-1源巨噬／泡沫细胞内胆固醇的含量,ADMA为30μmol/L和24h时作用最明显。 结论1.ADMA能够增加人THP-1源巨噬细胞吞噬脂质的能力和细胞内胆固醇的含量,呈浓度-时间依赖关系。2.ADMA能引起典型的THP-1源泡沫细胞形态学特征,增加人THP-1源泡沫细胞内胆固醇的含量,促进胆固醇在细胞内聚积,增加泡沫细胞的形成。3.ADMA可能是增加THP-1源巨噬细胞和泡沫细胞形成的重要因素,有可能成为临床防治动脉粥样硬化药物作用的新靶点。
[Abstract]:Aim ADMA is a potent endogenous nitric oxide synthase (NOS) inhibitor and is considered to be a new risk factor for cardiovascular disease. Monocytes / macrophages and their foam cells play an important role in the development of atherosclerosis. The purpose of this study was to investigate the effect of ADMA on cholesterol content in human THP-1-derived macrophages and foam cells. Methods Human THP-1 monocytes were cultured in vitro and differentiated into macrophages induced by phorbol ester (PMA). THP-1-derived macrophages were identified by immunohistochemistry. THP-1-derived macrophages were induced to differentiate into THP-1-derived foam cells by oxidized low density lipoprotein (ox-LDL). The THP-1-derived foam cells were identified by oil red O staining. The cholesterol content in THP-1-derived macrophages / foam cells was determined by enzyme-coupled colorimetry at different concentrations of ADMA (3.75, 7.5, 15,30 渭 mol / L) for different time (6,12,24 hours). Results Immunohistochemistry showed that CD68,THP-1 cells expressed in 85%THP-1 monocytes were induced to be THP-1-derived macrophages after induction of phorbol ester. The results of oil red O staining showed that a large number of lipid droplets existed in the cells, and THP-1-derived macrophages engulfed ox-LDL to form THP-1-derived foam cells. The results of enzyme-coupled colorimetry showed that ADMA increased cholesterol content in THP-1-derived macrophages / foam cells in a concentration-time dependent manner in vitro, and the effect was most obvious at ADMA of 30 渭 mol / L and 24 h. Conclusion 1.ADMA can increase the ability of human THP-1-derived macrophages to phagocytosis lipid and the content of intracellular cholesterol in a concentration-time dependent manner. 2. ADMA can induce typical morphological characteristics of THP-1-derived foam cells. 3. ADMA may be an important factor to increase the formation of THP-1-derived macrophages and foam cells, and increase the content of cholesterol in THP-1-derived foam cells, promote the accumulation of cholesterol in the cells, and increase the formation of foam cells. It is possible to become a new target of clinical anti-atherosclerotic drugs.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363
本文编号：2452111
[Abstract]:Aim ADMA is a potent endogenous nitric oxide synthase (NOS) inhibitor and is considered to be a new risk factor for cardiovascular disease. Monocytes / macrophages and their foam cells play an important role in the development of atherosclerosis. The purpose of this study was to investigate the effect of ADMA on cholesterol content in human THP-1-derived macrophages and foam cells. Methods Human THP-1 monocytes were cultured in vitro and differentiated into macrophages induced by phorbol ester (PMA). THP-1-derived macrophages were identified by immunohistochemistry. THP-1-derived macrophages were induced to differentiate into THP-1-derived foam cells by oxidized low density lipoprotein (ox-LDL). The THP-1-derived foam cells were identified by oil red O staining. The cholesterol content in THP-1-derived macrophages / foam cells was determined by enzyme-coupled colorimetry at different concentrations of ADMA (3.75, 7.5, 15,30 渭 mol / L) for different time (6,12,24 hours). Results Immunohistochemistry showed that CD68,THP-1 cells expressed in 85%THP-1 monocytes were induced to be THP-1-derived macrophages after induction of phorbol ester. The results of oil red O staining showed that a large number of lipid droplets existed in the cells, and THP-1-derived macrophages engulfed ox-LDL to form THP-1-derived foam cells. The results of enzyme-coupled colorimetry showed that ADMA increased cholesterol content in THP-1-derived macrophages / foam cells in a concentration-time dependent manner in vitro, and the effect was most obvious at ADMA of 30 渭 mol / L and 24 h. Conclusion 1.ADMA can increase the ability of human THP-1-derived macrophages to phagocytosis lipid and the content of intracellular cholesterol in a concentration-time dependent manner. 2. ADMA can induce typical morphological characteristics of THP-1-derived foam cells. 3. ADMA may be an important factor to increase the formation of THP-1-derived macrophages and foam cells, and increase the content of cholesterol in THP-1-derived foam cells, promote the accumulation of cholesterol in the cells, and increase the formation of foam cells. It is possible to become a new target of clinical anti-atherosclerotic drugs.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363
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