【摘要】：目的：研究顯示PAPP-A通過IGF-1激活PI3-K/Akt信號(hào)通路發(fā)揮促動(dòng)脈粥樣硬化（As）發(fā)生發(fā)展的作用，此外IGF-1還可通過MAPK信號(hào)通路促進(jìn)As發(fā)生發(fā)展。本文觀察PAPP-A對THP-1巨噬細(xì)胞源性泡沫細(xì)胞ABCA1、ABCG1和SR-B1表達(dá)及其介導(dǎo)膽固醇流出的影響，并探討其相關(guān)機(jī)制。 方法：THP-1巨噬細(xì)胞源性泡沫細(xì)胞經(jīng)不同濃度PAPP-A（0，10，50，250ng/mL）處理或者用250ng/mL PAPP-A共處理不同時(shí)間（0，6，12，24h）;運(yùn)用液體閃爍計(jì)數(shù)器檢測細(xì)胞內(nèi)膽固醇流出和磷脂流出，高效液相色譜分析細(xì)胞內(nèi)總膽固醇、游離膽固醇和膽固醇酯含量,實(shí)時(shí)熒光定量PCR和Western blot檢測ABCA1、ABCG1、SR-B1和LXRα、IGFBP-5mRNA和蛋白表達(dá)情況。以PI3-K抑制劑或MAPK阻斷劑或轉(zhuǎn)染IGF-1R siRNA對PAPP-A作用下的細(xì)胞共處理24小時(shí)，觀察對ABCA1、ABCG1、SR-B1及LXRα表達(dá)的影響。250ng/mL PAPP-A處理THP-1巨噬細(xì)胞源性泡沫細(xì)胞24h，ELISA檢測培養(yǎng)液中IGF-1的濃度，，實(shí)時(shí)熒光定量PCR檢測細(xì)胞內(nèi)IGF-1mRNA表達(dá)情況，Western blot檢測PAPP-A對PI3-K和Akt磷酸化水平的影響 結(jié)果：PAPP-A抑制ABCA1、ABCG1和SR-B1的表達(dá)及其介導(dǎo)的膽固醇流出和磷脂流出；LXRα介導(dǎo)了PAPP-A抑制ABCA1、ABCG1和SR-B1的表達(dá)以及細(xì)胞內(nèi)膽固醇流出和磷脂流出；PAPP-A通過IGF-1/PI3K/Akt信號(hào)通路抑制LXRα、ABCA1、ABCG1和SR-B1mRNA和蛋白的表達(dá)；THP-1巨噬細(xì)胞源性泡沫細(xì)胞中PAPP-A可激活I(lǐng)GF-1/PI-3K/Akt信號(hào)途徑。 結(jié)論：PAPP-A通過IGF-1/PI3K/Akt信號(hào)通路抑制LXRα表達(dá)，進(jìn)而抑制ABCA1、ABCG1和SR-B1表達(dá)及細(xì)胞內(nèi)膽固醇流出，從而發(fā)揮促As發(fā)生發(fā)展的作用。
[Abstract]:Aim: to investigate the role of PAPP-A in promoting the development of atherosclerotic (As) through IGF-1 activation of PI3-K/Akt signaling pathway, and IGF-1 can also promote the development of As through MAPK signaling pathway. The effects of PAPP-A on the expression of ABCA1,ABCG1 and SR-B1 in THP-1 macrophage derived foam cells and their effects on cholesterol efflux were investigated. Methods: THP-1 macrophage derived foam cells were treated with different concentrations of PAPP-A (010 ~ 10g / mL) or co-treated with 250ng/mL PAPP-A for 24 h. Cholesterol efflux and phospholipid efflux were detected by liquid scintillation counter, total cholesterol, free cholesterol and cholesterol ester contents were analyzed by HPLC, ABCA1,ABCG1,SR-B1 and LXR 偽 were detected by real-time fluorescence quantitative PCR and Western blot. Expression of IGFBP-5mRNA and protein. The effects of PI3-K inhibitor or MAPK blocker or IGF-1R siRNA transfection on the expression of ABCA1,ABCG1,SR-B1 and LXR 偽 in the cells treated with PAPP-A for 24 hours were observed. 250ng/mL PAPP-A treatment of THP-1 macrophage derived foam cells for 24 h. Detection of IGF-1 concentration in culture medium by ELISA and IGF-1mRNA expression in cells by real-time fluorescence quantitative PCR, Western blot effect of PAPP-A on PI3-K and Akt phosphorylation results: PAPP-A inhibits ABCA1, The expression of ABCG1 and SR-B1 and their mediated cholesterol and phospholipid efflux; LXR 偽 mediated PAPP-A to inhibit the expression of ABCA1,ABCG1, SR-B1, cholesterol and phospholipid, and PAPP-A inhibited the expression of LXR 偽, ABCA1,ABCG1, SR-B1mRNA and protein through IGF-1/PI3K/Akt signaling pathway. PAPP-A can activate IGF-1/PI-3K/Akt signaling pathway in THP-1 macrophage derived foam cells. Conclusion: PAPP-A inhibits the expression of LXR 偽, ABCA1,ABCG1 and SR-B1 and cholesterol efflux through IGF-1/PI3K/Akt signaling pathway, thus promoting the development of As.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363

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