本文選題：Ox-LDL + 誘導(dǎo)　； 參考：《浙江大學(xué)》2006年博士論文

【摘要】：動脈粥樣硬化(Atherosclerosis,AS)是一種多因素疾病,與多種危險因子包括血脂紊亂、糖尿病、肥胖和高血壓有關(guān),重要的基本病理改變是氧化低密度脂蛋白(oxidized low-density lipoprotein,Ox-LDL)在血管壁的積聚,其中巨噬細(xì)胞在血管壁的脂質(zhì)代謝中起著關(guān)鍵而復(fù)雜的作用。在早期脂質(zhì)浸潤中,巨噬細(xì)胞通過清道夫受體(CD36等)攝取Ox-LDL,形成泡沫細(xì)胞,大量積聚的Ox-LDL引起細(xì)胞毒性使細(xì)胞產(chǎn)生大量炎癥因子,改變了細(xì)胞的形態(tài)和信號轉(zhuǎn)導(dǎo),進(jìn)而損傷細(xì)胞影響動脈粥樣硬化的發(fā)展。因此,早期AS最基本的病理特征之一就是巨噬源泡沫細(xì)胞的形成和損傷,但目前其形成和損傷的時間和空間動力學(xué)機(jī)理仍不清楚。 本研究以PMA誘導(dǎo)人單核細(xì)胞U937源的巨噬細(xì)胞為靶細(xì)胞,通過Ox-LDL誘導(dǎo)的巨噬細(xì)胞源泡沫細(xì)胞模型,利用流式細(xì)胞術(shù)、激光共聚焦顯微術(shù)和分子生物學(xué)技術(shù)等實(shí)驗(yàn)技術(shù),結(jié)合動力學(xué)分析方法,研究了動脈粥樣硬化早期巨噬細(xì)胞源泡沫細(xì)胞形成和損傷的相關(guān)動力學(xué)機(jī)制。 本研究分別在Ox-LDL急性和慢性刺激等不同條件下,對影響泡沫細(xì)胞形成的部分關(guān)鍵因子膽固醇酯(CE)、總膽固醇(TC)、CD36、CD54和F-actin,以及[Ca~(2+)]i等進(jìn)行了時間和空間上的相關(guān)動力學(xué)研究。研究結(jié)果顯示,1)在慢性刺激下,Ox-LDL能顯著以時間依賴的方式誘導(dǎo)細(xì)胞內(nèi)CE和TC的升高(P0.01),尤其在24h時(CE/TC)%50%,表明了Ox-LDL誘導(dǎo)早期泡沫細(xì)胞形成的時間動力學(xué)變化,即Ox-LDL刺激24h后形成典型巨噬細(xì)胞源泡沫細(xì)胞;2)在急性和慢性刺激下,Ox-LDL能顯著誘導(dǎo)[Ca~(2+)]i的升高,但在急性刺激時,胞外液無論在有無Ca~(2+)存在的情況下,[Ca~(2+)]i都呈不同程度的升高,且在空間上形成明顯的跨膜Ca~(2+)梯度變化;3)在慢性刺激下,Ox-LDL以時間依賴的方式誘導(dǎo)細(xì)胞表面CD54和CD36表達(dá)的升高,以及F-actin時間和空間上多聚化的改變; 另外,本研究還分別在Ox-LDL急性和慢性刺激等不同條件下,對影響泡沫細(xì)胞內(nèi)線粒體損傷的部分關(guān)鍵因子H_2O_2、NO和O_2~(·-)、線粒體呼吸、線粒體膜電位(△ψm)、谷胱甘肽(GSH)和Bcl-2蛋白的表達(dá)、以及細(xì)胞的凋亡和壞死等進(jìn)行了相關(guān)動力學(xué)研究。研究結(jié)果顯示,1)在急性和慢性刺激下,Ox-LDL能顯著
[Abstract]:Atherosclerotic atherosclerosis is a multifactorial disease associated with multiple risk factors including dyslipidemia, diabetes, obesity and hypertension. An important underlying pathological change is the accumulation of oxidized low density lipoprotein (low-density) protein Ox-LDLs in the vascular wall. Macrophages play a key and complex role in lipid metabolism in vascular walls. In the early stage of lipid infiltration, macrophages ingest Ox-LDLthrough scavenger receptor CD36 and form foam cells. The accumulation of Ox-LDL leads to the production of a large number of inflammatory factors and changes the morphology and signal transduction of the cells. And then damage the cells to affect the development of atherosclerosis. Therefore, one of the most basic pathological features of early as is the formation and injury of macrophage foam cells, but the temporal and spatial dynamic mechanism of the formation and injury is still unclear. In this study, macrophages derived from human monocyte U937 induced by PMA were used as target cells, and Ox-LDL induced macrophage foam cell model was used. Flow cytometry, laser confocal microscopy and molecular biology techniques were used. The kinetic mechanism of macrophage-derived foam cell formation and injury in early atherosclerosis was studied by means of kinetic analysis. In this study, under different conditions of acute and chronic stimulation of Ox-LDL, some key factors affecting foam cell formation, such as cholesterol ester, total cholesterol (TCC) CD36, CD54 and F-actini, and [Ca~(2] I, were studied in time and space respectively. The results showed that Ox-LDL could significantly induce the increase of CE and TC in cells in a time-dependent manner, especially at 24 h, which indicated that Ox-LDL induced early foam cell formation in a time-dependent manner. That is to say, after 24 hours of Ox-LDL stimulation, typical macrophage derived foam cells were formed. Ox-LDL significantly induced the increase of [Ca~(2] I under acute and chronic stimulation, but in acute stimulation, [Ca~(2] I increased in varying degrees, regardless of the presence of Ca~(2 in the extracellular fluid. Furthermore, a significant transmembrane Ca~(2 gradient change was formed in space. Under chronic stimulation, Ox-LDL induced the increase of CD54 and CD36 expression on the surface of cells in a time-dependent manner, as well as the changes of F-actin in time and space. In addition, under different conditions such as acute and chronic Ox-LDL stimulation, the expression of H _ 2O _ 2NO and O _ 2C, mitochondrial respiration, mitochondrial membrane potential, glutathione (GSH) and Bcl-2 protein in foam cells were also studied. The kinetics of apoptosis and necrosis were studied. The results showed that 1) Ox-LDL was significant in acute and chronic stimuli.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2006
【分類號】：R363

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 李帥;NO/PKG對THP-1巨噬細(xì)胞ABCA1基因mRNA表達(dá)和膽固醇含量的影響[D];桂林醫(yī)學(xué)院;2011年

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本文編號：1830405