本文關(guān)鍵詞： 髓過(guò)氧化物酶 低密度脂蛋白 過(guò)氧化氫 單核巨噬細(xì)胞 阿司匹林　出處：《中南大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:觀察髓過(guò)氧化物酶(myeloperoxidase,MPO)對(duì)低密度脂蛋白(low density lipoprotein,LDL)的作用和對(duì)THP-1單核巨噬細(xì)胞泡沫化的影響和可能的機(jī)制。 方法:不同濃度的MPO(0.5、1、1.5、2、2.5u/L、)與LDL孵育;不同濃度MPO濃度(0.5、1、1.5、2、2.5u/L)加入過(guò)氧化氫(hydrogendioxide,H_2O_2)與LDL共孵育,TBA法檢測(cè)丙二醛(MDA)的生成量和瓊脂糖蛋白電泳檢測(cè)蛋白相對(duì)電泳率(REM)。在MPO-H_2O_2體系中加入不同濃度的阿司匹林(1、5、10mmol/1)與LDL共孵育后,TBA法檢測(cè)MDA的生成量和瓊脂糖蛋白電泳檢測(cè)蛋白相對(duì)電泳率。MPO途徑氧化生成的氧化型低密度脂蛋白(oxidized low densitylipoprotein,ox-LDL)與THP-1單核巨噬細(xì)胞共同培養(yǎng)24小時(shí)后,油紅O染色法檢測(cè)細(xì)胞的形態(tài)學(xué),液相色譜一質(zhì)譜聯(lián)用法檢測(cè)細(xì)胞內(nèi)膽固醇含量。 結(jié)果:1.不同濃度的MPO與LDL孵育后,與對(duì)照組(單純LDL)比較,MDA的生成量和蛋白相對(duì)電泳率(REM)差異均無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05);2.不同濃度MPO濃度加入H_2O_2后與LDL共孵育,與對(duì)照組比較,MDA生成量和REM均有明顯的增加,差異有統(tǒng)計(jì)學(xué)意義(P＜0.05);3.MPO-H_2O_2反應(yīng)體系中加入不同濃度的阿司匹林(1、5、10mmol/1)后與LDL共孵育,在阿司匹林濃度為1mmol/L,與對(duì)照組比較,MDA的生成量和REM差異均無(wú)明顯統(tǒng)計(jì)學(xué)意義(P＞0.05);阿司匹林濃度在5mmo/l、10mmol/L,與對(duì)照組比較,MDA的生成量和REM均明顯下降,差異有統(tǒng)計(jì)學(xué)意義(P＜0.05);4.MPO途徑氧化生成的ox-LDL與THP-1單核巨噬細(xì)胞共培養(yǎng)24小時(shí)后,油紅O染色形態(tài)學(xué)觀察;實(shí)驗(yàn)組巨噬細(xì)胞內(nèi)可見大量的紅色脂質(zhì)顆粒,正常對(duì)照組幾乎沒(méi)有紅色的脂質(zhì)顆粒。液相色譜—質(zhì)譜聯(lián)用法檢測(cè),總膽固醇(TC),游離膽固醇(FC),實(shí)驗(yàn)組細(xì)胞檢測(cè)CE/TC=52.70%(＞50%),對(duì)照組CE/TC=35.7%(＜50%)。 結(jié)論:1.MPO可能通過(guò)H_2O_2途徑介導(dǎo)LDL氧化并誘導(dǎo)THP-1單核巨噬細(xì)胞泡沫化。 2.5可司匹林可以抑制MPO-H_2O_2途徑介導(dǎo)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.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R363

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 劉恒道;髓過(guò)氧化物酶誘導(dǎo)內(nèi)皮細(xì)胞凋亡的分子機(jī)制[D];中南大學(xué);2012年

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本文編號(hào)：1527893