MPO对THP-1细胞泡沫化的影响和相关机制的实验研究

发布时间：2018-02-23 22:35

本文关键词： 髓过氧化物酶低密度脂蛋白过氧化氢单核巨噬细胞阿司匹林　出处：《中南大学》2008年硕士论文　论文类型：学位论文

【摘要】： 目的:观察髓过氧化物酶(myeloperoxidase,MPO)对低密度脂蛋白(low density lipoprotein,LDL)的作用和对THP-1单核巨噬细胞泡沫化的影响和可能的机制。方法:不同浓度的MPO(0.5、1、1.5、2、2.5u/L、)与LDL孵育;不同浓度MPO浓度(0.5、1、1.5、2、2.5u/L)加入过氧化氢(hydrogendioxide,H_2O_2)与LDL共孵育,TBA法检测丙二醛(MDA)的生成量和琼脂糖蛋白电泳检测蛋白相对电泳率(REM)。在MPO-H_2O_2体系中加入不同浓度的阿司匹林(1、5、10mmol/1)与LDL共孵育后,TBA法检测MDA的生成量和琼脂糖蛋白电泳检测蛋白相对电泳率。MPO途径氧化生成的氧化型低密度脂蛋白(oxidized low densitylipoprotein,ox-LDL)与THP-1单核巨噬细胞共同培养24小时后,油红O染色法检测细胞的形态学,液相色谱一质谱联用法检测细胞内胆固醇含量。结果:1.不同浓度的MPO与LDL孵育后,与对照组(单纯LDL)比较,MDA的生成量和蛋白相对电泳率(REM)差异均无统计学意义(P＞0.05);2.不同浓度MPO浓度加入H_2O_2后与LDL共孵育,与对照组比较,MDA生成量和REM均有明显的增加,差异有统计学意义(P＜0.05);3.MPO-H_2O_2反应体系中加入不同浓度的阿司匹林(1、5、10mmol/1)后与LDL共孵育,在阿司匹林浓度为1mmol/L,与对照组比较,MDA的生成量和REM差异均无明显统计学意义(P＞0.05);阿司匹林浓度在5mmo/l、10mmol/L,与对照组比较,MDA的生成量和REM均明显下降,差异有统计学意义(P＜0.05);4.MPO途径氧化生成的ox-LDL与THP-1单核巨噬细胞共培养24小时后,油红O染色形态学观察;实验组巨噬细胞内可见大量的红色脂质颗粒,正常对照组几乎没有红色的脂质颗粒。液相色谱—质谱联用法检测,总胆固醇(TC),游离胆固醇(FC),实验组细胞检测CE/TC=52.70%(＞50%),对照组CE/TC=35.7%(＜50%)。结论:1.MPO可能通过H_2O_2途径介导LDL氧化并诱导THP-1单核巨噬细胞泡沫化。 2.5可司匹林可以抑制MPO-H_2O_2途径介导的LDL氧化。
[Abstract]:Aim: to investigate the effects of myeloperoxidase (MPO) on low density lipoprotein (density) and the foaming of THP-1 mononuclear macrophages. Methods: LDL was incubated with different concentrations of MPO 0.5U / L ~ (-1) ~ (2.5) U / L ~ (-1); Different concentration of MPO: 0.5 ~ 1.5U / L) add hydrogen peroxide to Dioxide-H2O-2) Co-incubate with LDL to detect malondialdehyde (MDA) production and agarose protein electrophoresis to detect protein relative electrophoretic rate (REM). Add different concentrations of Aspirin to MPO-H_2O_2 system (10 mmol / 1) and LDL co-incubated with LDL. After incubation, the production of MDA was detected by TBA and the relative electrophoretic rate of protein by agarose protein electrophoresis. The oxidized low density lipoprotein oxidized lipoprotein (ox-LDL) was co-cultured with THP-1 mononuclear macrophages for 24 hours. The morphology of cells was detected by oil red O staining and cholesterol content was detected by liquid chromatography-mass spectrometry. Results 1. After incubating with LDL at different concentrations, there was no significant difference in MPO production and relative protein electrophoresis rate (REM) between LDL and control group (P > 0.05). LDL was co-incubated with H _ 2O _ 2 at different concentrations of MPO after addition of H _ 2O _ 2. Compared with the control group, the amount of malondialdehyde (MDA) and the amount of REM increased significantly (P < 0.05) and the difference was statistically significant (P < 0.05). The LDL was co-incubated with LDL after adding different concentrations of Aspirin in the system of H2O2 reaction. When the concentration of aspirin was 1 mmol / L, there was no significant difference in the amount of MDA and REM between the control group and the control group (P > 0.05), but the concentration of aspirin decreased significantly at 5 mmol / L 10 mmol / L, compared with the control group, the amount of MDA and REM decreased significantly. The difference was significant (P < 0.05). After co-culture of ox-LDL and THP-1 mononuclear macrophages for 24 hours, the morphology of oil red O staining was observed, and a large number of red lipid granules were observed in macrophages of experimental group. In the normal control group, there were almost no red lipid granules. Total cholesterol (TC) was detected by liquid chromatography-mass spectrometry (LC-MS), and free cholesterol (FCN) was detected. In the experimental group, CE-T / C (52.70%) was detected (> 50%), while in the control group, CE-T / C was 35.7g (< 50%). Conclusion: 1. MPO may mediate LDL oxidation and induce THP-1 mononuclear macrophage foaming through H _ S _ 2O _ 2 pathway. 2. 5 Costepidine inhibited LDL oxidation mediated by MPO-H_2O_2 pathway.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363

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