【摘要】： 目的ADMA是一種強(qiáng)烈的內(nèi)源性一氧化氮合酶(NOS)抑制劑,被認(rèn)為是心血管疾病的一個(gè)新的危險(xiǎn)因子。單核細(xì)胞／巨噬細(xì)胞及其形成的泡沫細(xì)胞在動(dòng)脈粥樣硬化的發(fā)生發(fā)展中具有重要作用。本研究探討ADMA對(duì)人THP-1源巨噬細(xì)胞和泡沫細(xì)胞內(nèi)膽固醇含量的影響。 方法體外培養(yǎng)人THP-1單核細(xì)胞,給予佛波酯(PMA)誘導(dǎo)分化成巨噬細(xì)胞,用免疫組化法鑒定THP-1源巨噬細(xì)胞；THP-1源巨噬細(xì)胞給予氧化低密度脂蛋白(ox-LDL)誘導(dǎo)分化為T(mén)HP-1源泡沫細(xì)胞,用油紅O染色鑒定THP-1源泡沫細(xì)胞。用酶偶聯(lián)比色法檢測(cè)不同濃度ADMA(3.75、7.5、15、30μmol/L)作用不同時(shí)間(6、12、24h)THP-1源巨噬／泡沫細(xì)胞內(nèi)膽固醇含量。 結(jié)果免疫組化結(jié)果表明佛波酯誘導(dǎo)后85%THP-1單核細(xì)胞表達(dá)CD68，THP-1細(xì)胞被誘導(dǎo)為T(mén)HP-1源巨噬細(xì)胞；油紅O染色結(jié)果顯示細(xì)胞內(nèi)有大量脂滴存在,THP-1源巨噬細(xì)胞吞噬ox-LDL形成THP-1源泡沫細(xì)胞。酶偶聯(lián)比色法結(jié)果顯示ADMA在體外以濃度-時(shí)間依賴(lài)的方式增加THP-1源巨噬／泡沫細(xì)胞內(nèi)膽固醇的含量,ADMA為30μmol/L和24h時(shí)作用最明顯。 結(jié)論1.ADMA能夠增加人THP-1源巨噬細(xì)胞吞噬脂質(zhì)的能力和細(xì)胞內(nèi)膽固醇的含量,呈濃度-時(shí)間依賴(lài)關(guān)系。2.ADMA能引起典型的THP-1源泡沫細(xì)胞形態(tài)學(xué)特征,增加人THP-1源泡沫細(xì)胞內(nèi)膽固醇的含量,促進(jìn)膽固醇在細(xì)胞內(nèi)聚積,增加泡沫細(xì)胞的形成。3.ADMA可能是增加THP-1源巨噬細(xì)胞和泡沫細(xì)胞形成的重要因素,有可能成為臨床防治動(dòng)脈粥樣硬化藥物作用的新靶點(diǎn)。
[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.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R363

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